Upload
others
View
1
Download
0
Embed Size (px)
Citation preview
19th Annual Upstate New York
Immunology Conference
Major Corporate Sponsors:
BD Biosciences
BioLegend, Inc.
Keynote Speakers:
Wayne M. Yokoyama, M.D.
Thomas J. Braciale, M.D., Ph.D.
Workshops by:
Dr. Yokoyama, Dr. Braciale, and Members of NIH
Panel Discussion:
Careers Outside of Academia
AAI Young Investigator Awards
eBioscience/affymetrix Trainee Travel Awards
Welcome to
The Sagamore Resort
and Conference Center
Bolton Landing, NY
October 24-27, 2016
Supported by NIH/NIAID
Grant Funding
2.
3.
Conference and Venue ............................................................................... 5
Schedule of Events .................................................................................... 6
Memories from 2015 .................................................................................. 15
Platinum Corporate Sponsors
BD Biosciences ..................................................................................... 16
BioLegend, Inc. .................................................................................... 18
Silver Plus Corporate Sponsors
eBioscience/affymetrix .......................................................................... 20
Krackeler Scientific .............................................................................. 22
Silver Corporate Sponsors .......................................................................... 24
Agilent Technologies ............................................................................ 25
EMD Millipore ...................................................................................... 26
Shenandoah ......................................................................................... 27
ThermoFisher ....................................................................................... 28
Bronze Corporate Sponsors ........................................................................ 29
American Association of Immunologists ....................................................... 30
NYIC Scientific Advisory Board .................................................................... 31
Institutional Financial Supporters ................................................................ 32
Grant Support ........................................................................................... 33
Keynote Speaker—Sponsored by BD Biosciences
Wayne M. Yokoyama, M.D. ................................................................... 34
Symposium I: Current Topics in Immunoregulation ...................................... 35
Symposium II: Neonatal Immunity .............................................................. 39
Oral Poster Presentations
Session A: Lymphocyte Immunobiology I ............................................... 42
Session B: Tumor Immunology .............................................................. 48
Table of Contents
4.
Corporate Presentation: BioLegend, Inc. ...................................................... 54
Oral Poster Presentations
Session C: Lymphocyte Immunobiology II .............................................. 55
Session D: Immunoregulation ................................................................ 61
Workshop I - Dr. Wayne Yokoyama
“Opportunities to Study the Immunology of Human Diseases” ................... 67
Corporate Presentation: BD Biosciences ....................................................... 68
Symposium III: Cancer Immunity ............................................................... 69
Symposium IV: Immune Mediated Disease .................................................. 72
Industry Panel
“Careers Outside of Academia” ............................................................. 75
Workshop II - Dr. Thomas Braciale
“The Business End of Academic Research” .............................................. 76
Workshop III—NIH Members
“Grantsmanship, Funding, and Mock Peer Review” ................................. 77
Symposium V: Immunity in Infectious Disease ............................................. 78
Keynote Speaker—Sponsored by BioLegend
Dr. Thomas J. Braciale .......................................................................... 82
Poster Listing ............................................................................................ 83
Poster Abstracts ........................................................................................ 84
Attendee Contact Information ..................................................................... 133
Author Index ............................................................................................. 139
5.
UPSTATE NEW YORK IMMUNOLOGY CONFERENCE (NYIC)
We’ve come a long way from Garnet Hill! This meeting started in 1997 as a small retreat to facilitate
interactions among young scientists, institutions, and renowned experts in the field of Immunology. In
just a few short years, the number of attendees grew and a larger venue was needed to meet the future
needs of the Conference.
We are happy to announce the American Association of Immu-
nologists (AAI) is once again providing 10 Young Investigator
Awards. eBioscience/affymetrix is also proving 10 Trainee Travel
Awards. All award winners will give Oral Poster Presentations.
There will also be two Workshops. Keynote speakers are Dr. Wayne M. Yokoyama
(Washington University) and Dr. Thomas J. Braciale (University of Virginia). As part of
our leisure activities, there will be a cruise on Lake George, as well as a recreational night including min-
iature golf, Wii, X-box, whiffle ball, and movies! Caldwell’s and Mr. Brown’s Pub will be open Wednes-
day night for informal discussions. Trainees will also have an opportunity to win an iPad during one of
two drawings. You must be present at the drawing to win!
While all these elements lend to the atmosphere, one simple principle goal of this Conference re-
mains. To provide an opportunity for young and senior scientists to gather in a setting that is diverse
enough to meet the needs of all attendees while remaining small enough to allow for personal interac-
tions. While always challenging, it is the goal of the NYIC Scientific Advisory Board and the NYIC Confer-
ence Organizers to give graduate students and postdoctoral fellows the opportunity to present their re-
search and engage in conversations that will stimulate further discussions, collaborations, and interest in
pursuing a new or different way of looking at their research.
We hope you share our enthusiasm and enjoy your time with us!
THE SAGAMORE RESORT
The Sagamore Resort and Conference Cen-
ter celebrated it’s 125 year anniversary in
2008. Since that time, the resort has gone
through some remarkable renovations. If this is
your first visit to the Resort, take some time to
enjoy the beauty that surrounds you. There
are many breath-taking views to be seen.
The staff are friendly, courteous, and hard-working. If you require any information or have a special
need, please see either Dawn Bellville, Administrative Coordinator for NYIC, or any of the Resort person-
nel. Many thanks to Lori Rehm (Director of Sales), Shelly Yeager (National Sales Manager), Derrick Ham-
mond (Conference Services Manager), Don Vilmar (Banquet Manager), Joel Clark (Function Set-up Man-
ager) and his amazing crew, CMI Communications-Audio/Visual Technician, Glen, along with all of the
associates who attend to our many needs. Thank you!
6.
Upstate New York Immunology Conference
Schedule of Events
Monday, October 24th
3:00-5:00 p.m. Hotel Check-in (Main Hotel Lobby) 3:00-5:00 p.m. Conference Registration (Albenia—Conference Center) 5:00-6:00 p.m. Welcome Reception (Conference Center Lobby) 6:00-6:30 p.m. Dinner (Bellvue) 6:30 p.m. Welcome and Introductions
Keynote Presentation
Sponsored by BD Biosciences Introduction: Dr. Dennis W. Metzger
Wayne M. Yokoyama, M.D. Levin Professor of Medicine Investigator, Howard Hughes Medical Institute Washington University
“Tissue-resident Natural Killer Cells” 7:30 p.m. American Association of Immunologists Young Investigator Awards and eBioscience Travel Awards (Award Presentations & Photos) Immediately Following Awards Presentation—Meet and Greet Activities
(Recreation Center)
7.
Tuesday, October 25th 7:00-8:15 a.m. Breakfast at Leisure (Use voucher towards total cost) (La Bella Vita)
8:25-8:30 a.m. Morning Announcements (Nirvana)
8:30-10:00 a.m. Symposium I: Current Topics in Immunoregulation (Nirvana) Chair: Dr. Brent Berwin 8:30-9:00 Michael A. Lynes, Ph.D. (University of Connecticut)
“The Stress Protein Metallothionein: a Small Protein with a Large Immunomodulatory Role”
9:00-9:30 Qi Yang, Ph.D. (Albany Medical College) “Biology of Inflammatory Innate Lymphoid Cells” 9:30-10:00 Karin Schneider, Ph.D. (SUNY Upstate)
“Influence of Colony Stimulating Factors on Virus Infections of Monocyte Lineage Cells”
10:00-10:15 a.m. Break and Display Posters (Conference Center Foyer/Bellvue) 10:15-11:15 p.m. Symposium II: Neonatal Immunity (Nirvana) Chair: Dr. Gary Winslow 10:15-10:45 Brain Rudd, Ph.D. (Cornell University) “The Fate of Neonatal and Adult CD8+ T cells During Infection is Linked to Their Developmental Origin” 10:45-11:15 David A. Lawrence, Ph.D. (Wadsworth/NYSDOH)
“Maternal-placental-fetal Interactions Affecting Offspring Im-munity and Behavior”
11:15-11:30 a.m. Break and Display Posters (Conference Center Foyer/Bellvue) 11:30-12:45 p.m. Oral Poster Presentations (Albenia) Session A—Lymphocyte Immunobiology I
8.
11:30-12:45 p.m. Oral Poster Presentations (Albenia) Session A - Lymphocyte Immunobiology I Chairs: Drs. Margaret Bynoe and Brian Rudd
11:30-11:45 Oyebola Oyesola, M.S. (Roswell Park Cancer Institute)
“The Prostaglandin D2 Receptor CRTH2 Mediates Interleukin-33-elicited Group 2 Innate Lymphoid Cell Accumulation in Tissues” (5)
11:45-12:00 Weishan Huang, Ph.D. (Cornell University)
“ITK Signaling via IRF4 Regulates the Development and Function of Type 1 Regulatory T Cells” (6)
12:00-12:15 Christina M. Post, M.S. (University of Rochester)
“The Ancestral Environment Shapes Antiviral CD8+ T Cell Responses Across Generations” (11)
12:15-12:30 Jennifer Yates, Ph.D. (Wadsworth Center)
“Early IL-10 Signals Favor Regulatory B Cell Over Memory B Cell Development during Cognate iNKT Cell Help” (17)
12:30-12:45 Victoria L. DeVault, B.S. (University of Vermont)
“SLAMf6 Modulates the NKT Cell Death Threshold” (20) 11:30-12:45 p.m. Oral Poster Presentations (Evelley) Session B - Tumor Immunology Chairs: Drs. Michael Robek and Yasmin Thanavala 11:30-11:45 Kelly L. Singel, B.S. (Roswell Park Cancer Institute)
“A Novel Barrier to Endogenous Anti-Tumor Immunity: Ovarian Cancer Ascites-activated Neutrophils Suppress T Cell Proliferation in a Contact-dependent Mechanism” (4)
11:45-12:00 Colin A. Powers, Ph.D. (Roswell Park Cancer Institute) “Tumor-induced Myeloid-derived Suppressor Cells Act via Remote Control to Inhibit L-selectin-dependent Adaptive Im-munity in Lymph Nodes” (24)
9.
12:00-12:15 Adaobi Amobi, B.S. (Roswell Park Cancer Institute)
“Tumor-Derived Indoleamine 2,3- Dioxygenase Regulates Density of Tumor Infiltrating CD8+ T cells and Myeloid Derived Suppressor Cells in a Murine Model of Ovarian Cancer” (26)
12:15-12:30 Amy Ku, B.S. (Roswell Park Cancer Institute)
“Negative Impact of Myeloid-derived Suppressor Cells on CD8 Effector T Cell Trafficking Within the Tumor Microenvironment” (27)
12:30-12:45 Anand Sharda, B.A. (Roswell Park Cancer Institute)
“Pretreatment Peripheral Blood Monocyte Subset Signature is Predictive of Patient Response to Dendritic Cell Vaccination” (48)
1:00-1:30 p.m. Lunch Buffet (Wapanak)
1:30-2:00 p.m. Platinum Corporate Sponsor—BioLegend, Inc. (Wapanak) Patrick Murphy, Ph.D. 2:00-3:15 p.m. Oral Poster Presentations (Albenia) Session C - Lymphocyte Immunobiology II Chairs: Drs. Nicholas Mantis and Gary Winslow
2:00-2:15 Stephanie L. Schell, B.S. (Penn State College)
“Autoantigen Availability Determines the Innate Sensing Requirement during Self-Antigen Driven Germinal Center Responses in Autoimmunity” (13)
2:15-2:30 Kevin Kenderes, B.S. (SUNY Upstate Medical University)
“IgM Memory B Cells Reconstitute Multiple B Cell Lineages and Provide Protection” (22)
2:30-2:45 Adam Utley, B.S. (Roswell Park Cancer Institute) “CD28 Induces Metabolic Fitness in LLPCs through NFκB-Mediated Irf4 Expression and ROS-Dependent Survival” (23)
10.
2:45-3:00 Joel R. Wilmore, Ph.D. (University of Pennsylvania)
“Commensal Microbes Drive the Generation of Systemic IgA Responses” (38)
3:00-3:15 Amber Papillion, B.S. (SUNY Upstate Medical University)
“Regulation of IgM Memory B cell Pool Size by the Inhibitory receptor FcγRIIb” (44)
2:00-3:15 p.m. Oral Poster Presentations (Evelley) Session D - Immunoregulation Chairs: Drs. Eyal Amiel and William O’Connor 2:00-2:15 Scott B. Minchenberg, B.S. (SUNY Upstate Medical University)
“Cytokine-mediated Regulation of Oligodendrocyte Metabolism” (8)
2:15-2:30 Camila Rosat Consiglio, M.S. (Roswell Park Cancer Institute) “Inflammation, Androgens and Macrophages in the Prostate:
Are We Missing the Link?” (18) 2:30-2:45 Phyu Thwe, B.S. (University of Vermont)
“Cell-Intrinsic Glycogen Metabolism Supports Early Activation and Maintains Metabolite Homeostasis in Dendritic Cells” (21)
2:45-3:00 Angelica Costello, B.A. (Albany Medical College)
“Macrophages Negatively Regulate Hematopoietic Stem Cells in Murine Aplastic Anemia” (40)
3:00-3:15 Travis Walrath, B.S. (Albany Medical College)
“Antagonistic Control of Intestinal Wnt Expression by IBD-Associated Cytokines” (49)
AAI Young Investigator Award and Oral Poster Presentation
eBioscience Trainee Travel Award and Oral Poster Presentation
3:15-3:30 p.m. Break 3:30-5:00 p.m. Workshop I - Dr. Wayne Yokoyama (Wapanak) “Opportunities to Study the Immunology of Human Diseases”
11.
5:00-6:30 p.m. Vendor/Poster Mixer (Bellvue) Poster Viewing and Questions (Odd Numbers) 6:30-7:00 p.m. Dinner (Wapanak)
7:00-7:30 p.m. Platinum Corporate Sponsor-BD Biosciences (Wapanak) Arielle Ginsberg 7:30-9:00 p.m. Vendor/Poster Mixer (Bellvue) Poster Viewing and Questions (Even Numbers) iPad drawing during this event. Must be present to win. 9:00-9:15 p.m. Remove Posters (Posters left behind will be discarded) Wednesday, October 26th 7:00-8:15 a.m. Breakfast at Leisure (Use voucher towards total cost) (La Bella Vita) 8:25-8:30 a.m. Morning Announcements (Nirvana)
8:30-9:30 a.m. Symposium III: Cancer Immunity (Nirvana) Chair: Dr. Edith Lord 8:30-9:00 Joseph Barbi, Ph.D. (Roswell Park) “Exploring the Mechanisms of Neurotrophin-mediated Immune Tolerance and Their Implications for Autoimmunity and Cancer” 9:00-9:30 Fiona Ginty, PhD. (GE Global) “In situ Analysis of Multiple Immune Cells in Tumor and Microenvironment” 9:30-10:30 a.m. Symposium IV: Immune Mediated Disease (Nirvana) Chair: Dr. Wayne Yokoyama
12.
9:30-10:00 Iwona Buskiewicz, Ph.D. (University of Vermont) “Reactive Oxygen Species Induce Virus-dependent MAVS-oligomerization in Systemic Lupus Erythematosus” 10:00-10:30 Timothy Chapman, Ph.D. (University of Rochester) “Increasing Regulatory Tone to Suppress Allergic Inflammatory Responses in the Lung” 10:30-10:45 a.m. Beverage Break (Conference Center Foyer)
10:45-11:45 a.m. Industry Panel (Albenia) “Careers Outside of Academia” 11:45-12:45 p.m. Lunch Buffet (Wapanak) 12:45 -2:00 p.m. Workshop II - Dr. Thomas Braciale (Nirvana) “The Business End of Academic Research” 2:15-2:30 p.m. Meet at Dock—Prepare to board The Morgan (Please have your boarding pass.)
2:30-4:00 p.m. Cruise sponsored by Krackeler Scientific, Inc. (Refreshments provided/Cash Bar)
4:00-6:00 p.m. Free Time 4:15-5:15 p.m. Scientific Advisory Board Meeting (Board Members Only) (Empire Room)
6:00-7:00 p.m. Dinner Buffet (Wapanak)
7:00-8:30 p.m. Workshop III—NIH Members “Grantsmanship, Funding, and Mock Peer Review”
8:30-11:30 p.m. Mr. Brown’s Pub and Caldwell’s Informal Discussions
13.
Thursday, October 27th
7:00-8:15 a.m. Breakfast at Leisure (Use voucher towards total cost)
(La Bella Vita)
8:25-8:30 a.m. Morning Announcements (Nirvana)
8:30-10:00 a.m. Symposium V: Immunity in Infectious Disease (Nirvana) Chair: Dr. Thomas Braciale
8:30-9:00 Richard I. Enelow, M.D. (Dartmouth College) “Control of the CD8+ Effector Population in Influenza Infection”
9:00-9:30 Christine King, Ph.D. (SUNY Upstate) “Mast Cells in Kaposi’s Sarcoma – A Model for the Pathogenesis of KSHV-driven Oncogenesis”
9:30-10:00 Ira J. Blader, Ph.D. (University at Buffalo) “Toxoplasma Infections of the Nervous System”
10:00-10:15 a.m. Beverage Break and Check-out (Conference Center Foyer/Hotel Lobby)
10:15-11:15 a.m. Keynote Presentation – Sponsored by BioLegend (Nirvana)
Introduction: Dr. Richard Enelow
Dr. Thomas J. Braciale Director, Emeritus Carter Immunology Center Professor of Pathology and Molecular Medicine University of Virginia
“Dendritic Cells in the Host Response to Stress or Dendritic
Cells Can Do More Than Present Antigens”
11:15-11:30 a.m. Closing Remarks(Nirvana) iPad drawing during this event. Must be present to win.
14.
Departure
Please join us next year for the
20th Annual
Upstate New York Immunology Conference
October 23-26, 2017
(Monday-Thursday)
The Sagamore Resort and Conference Center
Bolton Landing, NY
15.
NYIC 2015
16.
Platinum Corporate Sponsor
BD Biosciences
BD OptiBuild™ custom reagentsExactly the reagents you want, when you want them
The perfect conjugates to complete your artfully crafted panel
BD OptiBuild™ reagents give researchers access to custom antibody-dye combinations as easily and conveniently as ordering a standard catalog item. BD OptiBuild reagents are available in small, 50-microgram vial sizes and are ready to ship within hours of your order, usually arriving at your location in two to four days. If you’ve ever crafted a multicolor panel only to have to start over again because one or more of the conjugates you’ve specified is not available, you’ll truly appreciate BD OptiBuild reagents.
With BD OptiBuild reagents, your options are no longer limited to commonly available, in-stock products. Now you can complete your multicolor panels as you’ve designed them or simply explore and try new reagent combinations that might advance your research. Find your next conjugate at bdbiosciences.com/go/optibuild
For Research Use Only. Not for use in diagnostic or therapeutic procedures.© 2016 BD. BD, the BD Logo and all other trademarks are property of Becton, Dickinson and Company.23-18276-00
BD Life SciencesBiosciences2350 Qume DriveSan Jose, CA 95131bdbiosciences.com
18.
Platinum Corporate Sponsor
BioLegend, Inc.
World-Class Quality | Superior Customer Support | Outstanding Value
Toll-Free Tel: (US & Canada): 1.877.BIOLEGEND (246.5343)Tel: 858.768.5800
biolegend.com
08-0062-04
Learn more at: biolegend.com/� ex-t
MHC multimers that identify antigen-speci� c T cells have become the standard to measure speci� c T cell responses, from basic research to applied and clinical research. Flex-T™, BioLegend’s technology to study antigen-speci� c T cells, o� ers the unique advantage of custom peptide loading using a UV-dependent peptide exchange. The system also allows for two color combinatorial barcoding, making it possible to detect a diverse set of antigen-speci� c T cells with a single small volume of sample.
Flex-T™ Unparalleled MHC Multimer Versatility
BioLegend is ISO 9001:2008 and ISO 13485:2003 Certi� ed
A) CD8+ T cells previously gated on lymphocytes (FSC vs SSC) and 7-AAD negative events, the exclusion cocktail contains Alexa Fluor® 700 anti-CD4, CD19, CD14, and CD16. B) Antigen-speci� c CD8+ T cells, gated as described, detected with Flex-T™ tagged with PE and APC. HLA-A*11:01 Flex-T™ is loaded with EBV peptide (IVTDFSVIK).
The most advanced technology to � nd unique T cells
20.
Silver-Plus Corporate Sponsor
22.
Silver Plus Corporate Sponsor
We are proud to sponsor
The 19th Annual Upstate New YorkImmunology Conference 2016
Krackeler Scientific is a leading distributor of quality scientific products. We serve customers nationwide in biotechnology, nanotechnology,
life sciences, pharmaceutical, biomedical, environmental, & industrial sectors.
Our comprehensive product line represents all of the leading manufacturers including Corning, Sigma, Agilent, Eppendorf & Celltreat. Our home is Albany, NY.
www.krackeler.com • [email protected] • 800-334-7725
24.
Silver Corporate Sponsors
Aligent Biosciences
EMD Millipore
Shenandoah
ThermoFisher
Using as few as 15,000 live cells and an Agilent Seahorse XFp Analyzer,
metabolic phenotype as well as metabolic potential —
LEARN MORE
Extracellular Acidification Rate (ECAR)
Oxy
gen
Con
sum
ptio
n R
ate
(OC
R)
Glycolysis
Mito
chon
dria
l Res
pira
tion
Aerobic
Quiescent
Energetic
Glycolytic
BaselinePhenotype
StressedPhenotype
Metabolic Potentia
l
Agilent Seahorse XFp Cell Energy Phenotype Test Kit
Seahorse Biosci
ence is
Now Agilent Technologies
Flow Cytometers
Up to 3 lasers and 14 simultaneous detectionparameters
Particle detection down to 00.2 μmVersatile and user friendly softwareEasy operation and minimal maintenance
Benefits
No sheath fluid and absolute cell counts
Guava easyCyte™ Flow Cytometers
.
Benefits
MADE IN THE USAwww.shenandoah-bt.com
Start Your Test Drive Today:
Step 1: Order a Mini size of one of our many human, mouse, or rat cytokines to use in your lab- Just $60 for enough material to put through the paces and see for yourself why we are the Protein Pros!
Step 2: Order a larger size of the same protein within 3 months and receive a FULL CREDIT for the trial mini. Simply ask for the Test Drive Credit by phone, email or in the notes section of your online shopping cart. It’s just that simple!
Visit our website for more details.
CYTOKINES GROWTH FACTORS
CHEMOKINES INTERLEUKINS
The Highest Quality PROTEINS
at Super Competitive PRICES
29.
Bronze Corporate Sponsors
Tonbo Biosciences
30.
In recognition of the significance
of this meeting and work being done by
Graduate Students
and Postdoctoral Fellows,
the
American Association
Of Immunologists
has provided
Ten(10) Young Investigator Awards.
Each will receive a monetary award,
as well as the opportunity
to present their research both in poster format and brief talks.
31.
NYIC Scientific Advisory Board
Institutional Representatives
Albany Medical College
Jim Drake and Kate MacNamara
(NYIC Conference Organizers)
Cornell University
Margaret Bynoe
Dartmouth College
Brent Berwin
Roswell Park Cancer Institute
Yasmin Thanavala
SUNY Upstate Medical University
Gary Winslow
University at Buffalo
Beth Wohlfert
University of Connecticut
Steven Szcezpanek
University of Rochester Medical Center
Edith Lord
University of Vermont
Eyal Amiel
Wadsworth Center/SUNY Albany
Nicholas Mantis
32.
Institutional Financial Supporters
Albany Medical College
Alumni Association
Cornell University
Microbiology & Immunology
Dartmouth College
Department of Microbiology & Immunology
Roswell Park Cancer Institute
Department of Immunology
SUNY Upstate Medical University
Microbiology & Immunology Program
University at Buffalo
Buffalo School of Medicine
Department of Microbiology & Immunology
University of Connecticut
Department of Molecular & Cellular Biology
Center of Excellence for Vaccine Research
University of Rochester Medical Center
Department of Microbiology & Immunology
University of Vermont
Vermont Center for Immunology & Infectious Diseases
Wadsworth Center
33.
Grant support provided to
Graduate Students
and
Postdoctoral Fellows
by the
National Institutes of Health
National Institute of
Allergy and Infectious Diseases
R13AI051522
“Thank You”
34.
Keynote Speaker
Wayne M. Yokoyama, M.D. Levin Professor of Medicine
Investigator, Howard Hughes Medical Institute
Rheumatology Division, Dept. of Medicine
Washington University School of Medicine
St. Louis, Missouri
“Tissue-Resident Natural Killer Cells”
Most of what we know about natural killer (NK) cells comes from study of NK cells from the
mouse spleen and human peripheral blood. However, solid organs, such as the mouse liver, con-
tains two populations of NK cells, circulating conventional NK cells that resemble splenic NK cells,
and tissue-resident NK cells that do not recirculate. These NK cell populations can be distin-
guished by cell surface markers and transcription factor-dependence, indicating that they form
distinct lineages. Humans may also have tissue-resident NK cells but lineage-specific markers in
mice may not be easily translated to humans. Here I will describe our work on mouse tissue-
resident NK cells and progress we have made in characterizing human liver tissue-resident NK
cells.
Dr. Yokoyama’s research includes the role of NK cells in anti-pathogen defense led to studies of
innate immune responses to large DNA viruses, MCMV and CPXV. His lab is especially interested
in how these viruses interact with the host. They are also studying the interplay between innate
and adaptive immunity. They are also embarking on human studies aimed at understanding the
etiology and pathogenesis of autoimmune disease, especially rheumatic disorders (i.e., rheuma-
toid arthritis and Kawasaki disease.)
35.
Symposium I
Current Topics in Immunoregulation
Chair : Dr. Brent Berwin
36.
The Stress Protein Metallothionein: A Small Protein with a Large
Immunomodulatory Role
Michael A. Lynes
University of Connecticut, Storrs CT
Metallothionein (MT) is a protein that makes many interwoven contributions to cellular homeosta-
sis and to the management of cellular function under stressful conditions. These contributions include man-
aging essential metals such as copper and zinc, and regulating the local redox microenvironment, which
have been linked to MT’s influence on intracellular signaling cascades and many transcription factors (e.g.
Sp1 and NF-kB). MT can serve in these roles as a consequence of its thiol-rich nature: fully 33 mol% of
the protein is cysteine. MT has a complex upstream regulatory region, which is responsible for its expres-
sion as a consequence of cellular exposure to divalent heavy metal cations, reactive oxygen species, endo-
toxin, acute phase cytokines (IL-1, IL-6, and TNF-a), glucocorticoids, IFN-g, and aromatic hydrocarbons.
These inducers and regulatory roles suggest that MT plays an important role in the progression of an im-
mune response. We have shown that MT can influence elements of both the innate and adaptive immune
response. At least some of this effect is a consequence of its effect on intracellular NF-kB, while other
changes appear to rely on the MT-mediated management of intracellular Zn and the role that ion plays in
phosphatase activity. Under stressful circumstances, MT can exit the cell, despite the lack of a signal pep-
tide, and appears in extracellular fluids and spaces such as serum, urine, milk, bronchoalveolar lavage and
pancreatic acini, bronchoalveolar spaces, and liver sinusoids. We have shown that this extracellular pool
can serve as an activator of chemotactic responses, and that a targeted disruption of the Mt1 and Mt2 genes
or monoclonal antibody blockade of the extracellular pool of MT can significantly diminish the severity of
DSS-induced colitis in mice. Taken together, these observations suggest MT as an interesting and novel
target for therapeutic manipulation of the immune response.
37.
Biology of Inflammatory Innate Lymphoid Cells
Qi Yang and Kangning Zhang
Department of Immunology and Microbial Disease, Albany Medical College,
Albany, NY 12208
Mechanisms that control lymphocyte lineage stability and plasticity remain elusive. Recent work
indicates that innate lymphoid cells (ILC) possess substantial functional plasticity. Whereas natural type-2
innate lymphoid cells (nILC2) respond to IL-33 and produce type-2 cytokines, inflammatory ILC2 (iILC2)
respond to IL-25 and can co-produce both type-2 cytokines and also the ILC3-characteristic cytokine IL-
17. The transcriptional regulatory mechanisms that drive this plasticity, and the importance in health and
disease, remain unknown. Here we show that iILC2 are potent inducers of airway inflammation in re-
sponse to acute house-dust mite challenge. We find that Notch signaling drives the emergence of iILC2.
Acute blockade of Notch signaling abolished functional iILC2, but not nILC2, in vivo. Exposure of isolat-
ed nILC2 to Notch ligands in vitro altered their cytokine responsiveness and elicited dual IL-13/IL-17 pro-
duction, thus converting nILC2 into iILC2. In mature ILC2, Notch transcriptional complex bound to the
Rorc gene locus and promoted its expression, but did not affect the expression of Gata3, therefore confer-
ring ILC3-like capability without compromising primary ILC2 function. Together these results reveal a
novel role for Notch signaling in eliciting the plasticity of ILC2 and driving the emergence of highly pro-
inflammatory innate lymphocytes.
38.
Influence of Colony Stimulating Factors on Virus infections of Monocyte
Lineage Cells
Karin Schneider
SUNY Upstate Medical University
Monocyte lineage cells are common targets for infection and innate immune activation by many
pathogenic viruses. As both infection and innate activation are key determinants of viral pathogenesis espe-
cially in the central nervous system, an analysis of colony stimulating factors on these events was per-
formed in cultures infected with Theiler’s Murine Encephalomyelitis Virus (TMEV) or Zika virus (ZIKV).
Our results suggest that the two single-stranded, positive sense viruses can uniquely interact to different
monocyte lineage cells.
39.
Symposium II
Neonatal Immunity
Chair : Dr. Gary Winslow
40.
The Fate of Neonatal and Adult CD8+ T cells During Infection is Linked to Their
Developmental Origin
Jocelyn Wang1, Erin Wissink2, Neva Watson1, Norah L. Smith1, Arnold Reynaldi3,
Andrew Grimson2, Miles P. Davenport3, and Brian D. Rudd1
Department of Microbiology and Immunology1, Cornell University, Ithaca, NY 14853
Department of Molecular Biology and Genetics2, Cornell University, Ithaca, NY 14853
Kirby Institute for Infection and Immunity3, University of New South Wales,
Sydney, Australia
We recently showed that neonatal CD8+ T cells fail to become memory cells because of an inher-
ent propensity to rapidly proliferate and become terminally differentiated. However, the underlying basis
for these differences remained unclear. To determine whether neonatal CD8+ T cells behave differently
than adults because of age-related differences in T cell homeostasis or thymopoiesis, we directly compared
the behavior and transcriptomes of neonatal and adult CD8+ T cells that had undergone similar amounts of
homeostatic proliferation in the periphery or were at the same stage of thymic development. We also com-
pared T cell maturation by fetal and adult precursors in the adult thymus and examined whether fetal-
derived CD8+ T cells behave differently than their adult counterparts. Our data demonstrates that neonatal
and adult CD8+ T cells adopt different fates after infection because they are derived from distinct HSCs,
which express different amounts of Lin28b. We also developed a system to ‘timestamp’ CD8+ T cells in
situ at various stages of development (1d, 1wk, and 4wks) and examined their behavior at 8 wks of age.
These data indicates that the spectrum of CD8+ T cell differentiation observed after infection is influenced
by when the responding cells were initially made.
41.
Maternal-placental-fetal Interactions Affecting Offspring Immunity and Behavior
David A. Lawrence1,2, Tapan Mondal1, Alvaro Mendoza1, Jane Lubliner2, and
Lori Kearsing3 1 Wadsworth Center/New York State Department of Health,
2University at Albany School of Public Health, and 3Center for Disability Services
Luminex and grating coupled surface plasmon coupled fluorescence (GCSPCF) technologies were
used to screen samples from humans and mice. The human samples were obtained from newborn dried
blood spots (NDBSs) collected from babies born in New York and later diagnosed with autism spectrum
disorder (ASD) or typical development. The proteins eluted from the NDBSs were compared to plasma
protein differences from the BTBRT+ltpr3tf/J (BTBR) mouse strain with ASD-like behavior and the
C57BL/6J (B6) strain, which has normal behavior. BTBR mice have reduced social interactions, impaired
play, low exploratory behavior, unusual vocalizations and high anxiety; they also have substantial deple-
tion of the corpus callosum and severely reduced hippocampal commissure. Antibody specificities and iso-
types from the NDBSs of babies that developed ASD and those from BTBR mice were assessed by
GCSPCF and Luminex analysis and shown to differ from the antibody specificities and isotypes from hu-
mans and mice with typical development and usually normal behaviors. The human and mouse compari-
sons demonstrate that humans and mice with different behaviors have different antibody and cytokine pro-
files from the control populations. The human and mouse comparisons demonstrate that humans and mice
with different behaviors have different antibody and cytokine profiles from the control populations. Mater-
nal-placental-fetal interactions leading to heightened antenatal inflammation and oxidative stress are sug-
gested to affect the aberrant immunity and behavior of offspring.
42.
Oral Poster Presentations
Session A:
Lymphocyte Immunobiology I
Chairs: Drs. Margaret Bynoe
and Brian Rudd
43.
The Prostaglandin D2 Receptor CRTH2 Mediates Interleukin-33-elicited Group 2
Innate Lymphoid Cell Accumulation in Tissues
Oyebola Oyesola, Lauren M. Webb, Rebecca Cubitt, Elia Tait Wojno
Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca NY 14853, United
States
Group 2 innate lymphoid cells (ILC2s) are rare innate immune cells that contribute to the development of
type 2 inflammation. These cells and associated type 2 inflammation promote allergic lung inflammation, which
affects nearly 8% of the general population in the United States. Previous studies have shown that the accumulation
of ILC2s at inflammatory sites is mediated by epithelial cell-derived cytokines and alarmins, particularly interleukin-
33 (IL-33). In addition, prostaglandin D2 (PGD2), a bioactive lipid produced by mast cells, induces accumulation of
ILC2s at inflammatory sites by binding to its receptor chemoattractant receptor-homologous molecule expressed on
Th2 cells (CRTH2), which is expressed by ILC2s. However, whether the IL-33-IL-33 receptor (IL-33R) and PGD2-
CRTH2 pathways coordinate or intersect to regulate ILC2 responses during type 2 lung inflammation has not fully
been explored in vivo. In this study, we investigated the role of the PGD2-CRTH2 pathway during IL-33-elicited
ILC2 accumulation and type 2 inflammation in the lung. When wild-type mice were treated either systemically by
intraperitoneal injection or locally by intranasal injection with recombinant murine IL-33 (rmIL-33) to induce type 2
inflammation, ILC2 frequency and number increased in the lung parenchyma. In contrast, ILC2s accumulated in the
lung of mice deficient in CRTH2 (CRTH2KO) following intranasal but not intraperitoneal treatment with rmIL-33.
The defect in ILC2 accumulation in the CRTH2KO lung following systemic rmIL-33 treatment was not due to dif-
ferences in expression of the IL-33R, cell death, apoptosis, or cell proliferation. Therefore, our data suggest that the
PGD2-CRTH2 pathway acts downstream of IL-33 to partially mediate ILC2 accumulation in the lung during type 2
inflammation that originates in the periphery, acting via pathways other than those controlling cell responsiveness to
IL-33, death, or proliferation. Future studies will focus on determining if ILC2 accumulation in the lung in response
to IL-33 is mediated via CRTH2-dependent migration to or retention in the lung. A better understanding of how cy-
tokines and bioactive lipid mediators interact to regulate ILC2 responses will be important in informing the use and
development of drugs that target these pathways to treat ILC2-associated type 2 airway inflammation.
Poster 5
44.
ITK Signaling via IRF4 Regulates the Development and Function of Type 1
Regulatory T cells
Weishan Huang*, Sabrina Solouki, Nicholas Koylass, and Avery August*
Department of Microbiology and Immunology, Cornell University, Ithaca, NY 14853.
Type 1 regulatory T (Tr1) cells lack the expression of Foxp3 but have significant regulatory function in sup-
pressing inflammation and promoting tolerance, in part via their expression of the immunosuppressive cytokine IL-
10. Tr1 cells differentiate in response to signals engaging T cell receptor (TCR) and/or the regulatory cytokine mi-
lieu. The non-receptor tyrosine kinase ITK is a key modulator downstream of TCR, playing critical roles in T cell
development and function. Using mouse models carrying Foxp3RFP and IL-10GFP dual reporters, we found that, in
the absence of ITK, TCR activation-driven development of Foxp3- IL-10+ Tr1 cells is severely impaired in various
organs (spleen, blood, lung, gut, and fat). Itk-/- mice were also deficient in mounting a mucosal Tr1 cell response
during parasitic (Nippostrongylus Brasiliensis) and viral (Influenza A) infections.
Naïve Itk-/- thymic and splenic Foxp3- CD4+ T cells also exhibited severe deficiency in Tr1 differentiation
under Tr1 polarizing condition. Although Itk-/- CD4+ T cells proliferated under Tr1 differentiating conditions, they
failed to up-regulate IL-10, and Tr1 cell markers LAG3, CD49b, ICOS, PD-1, c-Maf, AHR, and IRF4 to levels ob-
served in WT cells, suggesting that ITK is critical for Tr1 cell fate programming. Utilizing a transgenic mouse model
carrying an allele sensitive mutant of ITK (ITKas) that allows ITK kinase specific blockade by a small molecule
3MB-PP1, we determined that the expression of the aforementioned markers, as well as the balance between IL-10
and IFN-g (gamma) production during Tr1 differentiation, are dependent on ITK kinase activity. Furthermore, using
cells from an ITKas-Foxp3RFP/IL-17AGFP dual reporter mouse model, we find that ITK kinase activity is required
for optimal Th17 trans-differentiation to Tr1 cells. We also find that inhibiting ITK kinase activity diminished Tr1
differentiation by human CD4+ T cells. The requirement for ITK function during Tr1 cell development can be re-
stored by the expression of the transcription factor IRF4. Finally, specifically targeting ITK kinase activity in already
differentiated Tr1 cells diminished their suppressive function.
We conclude that the TCR/ITK/IRF4 pathway is required for the development and function of Tr1 cells,
which may be targeted to modulate regulatory immunity for clinical benefit.
*Correspondence: Weishan Huang ([email protected]) and Avery August ([email protected]).
Poster 6
45.
The Ancestral Environment Shapes Antiviral CD8+ T cell Responses
Across Generations
Christina M. Post, Jason Myers, and B. Paige Lawrence
Department of Environmental Medicine, University of Rochester School of Medicine & Dentistry,
Rochester NY
Recent studies have linked health fates of great-grandchildren to environmental exposures of their great
grandparents. However, few studies have considered whether ancestral exposures influence the immune system
across generations. Here we report novel findings regarding transgenerational transmission of altered T cell respons-
es resulting from maternal (F0) exposure to an environmentally relevant aryl hydrocarbon receptor (AHR) ligand.
The AHR is a transcriptional regulator that plays diverse roles in cellular function, including modulating immune
responses. AHR ligands comprise several classes of pollutants, such as dioxins and PCBs, as well as molecules from
foods and other sources. AHR-binding pollutants cross the placenta and are excreted in breast milk. In animal mod-
els and human populations, early life exposure to dioxins and PCBs is associated with persistent defects in the off-
spring’s immune function. Using a mouse model, maternal exposure to the AHR ligand and pollutant 2,3,7,8-
tetrachlorodibenzo-p-dioxin (TCDD) results in a significantly reduced CD8+ T cell response to influenza A virus
(IAV) in the adult offspring (F1), compared to the response of infected offspring of control-treated dams. Specifical-
ly, there are significantly fewer cytotoxic T lymphocytes (CTL; CD44hiCD62Llo), virus-specific CD8+ T cells, and
CD8+ T cells that produce interferon gamma (IFNγ). Transcriptomic analyses using sorted CD8+ T cells from F1
offspring of TCDD and control dams support new evidence that triggering AHR during development changes pro-
gramming of senescence or exhaustion regulatory pathways. Follow up studies show increased expression of pro-
teins associated with hindered T cell responses, such as CTLA-4 and KLRG1 on CD8+ T cells. We next asked
whether the diminished CD8+ T cell response in the F1 generation was observed in the F3 generation. We detected
fewer CTL and virus-specific CD8+ T cells in the TCDD F3 lineage, as well as increased expression of CTLA-4 and
KLRG1 compared to control F3 lineage following IAV infection. These data indicate that F0 maternal exposure to
AHR ligands is capable of disrupting immune function not only via direct activation of the AHR in the F1 genera-
tion, but also by reprogramming immune responses in subsequent generations. This has broad implications for un-
derstanding how the environment of prior generations shapes susceptibility to pathogens and antiviral immunity in
later generations.
Poster 11
46.
Early IL-10 Signals Favor Regulatory B cell Over Memory B cell Development during Cognate
iNKT Cell Help
Jennifer Yates1,4, Emilie Vomhof-Dekrey1, Paula Lanthier1, Katja Mohrs1, Thomas Hägglöf 2,
Natacha Veerapen3, Gurdyal Besra3, Mikael Karlsson2, and Elizabeth Leadbetter1,5 1Trudeau Institute, Saranac Lake, NY 12983; 2Karolinska Institutet, Stockholm, Sweden;
3School of Biosciences University of Birmingham, Birmingham, UK; 4Wadsworth Center, NYSDOH, Albany, NY; 5University of Texas Health Science Center at San Antonio,
San Antonio, TX
Effective generation of humoral B cell memory is dependent upon help from CD4+ T cells. We and
others have found that invariant natural killer T (iNKT) cells can provide both cognate and non-cognate
helper signals to enhance B cell responses. While both cognate and non-cognate iNKT cell help induce
class-switched, antigen-specific humoral immune responses - only non-cognate iNKT cell help drives the
formation of humoral memory. Rather, cognate iNKT cell help drives an early, un-sustained expansion of
germinal center B cells and antigen-specific antibody production. Therefore, we posit that cognate help
provided to B cells by iNKT cells is fundamentally different from the help provided by conventional CD4+
T cells. We now find that glycolipid immunization drives considerable IL-10 transcription by many differ-
ent spleen cell populations including dendritic cells, plasmablasts, iNKT cells, and B regulatory cells. Cog-
nate iNKT cell help expands antigen-specific IL-10 producing B regulatory cells upon primary immuniza-
tion, and IL-10 producing iNKT10 cells following secondary antigen challenge. Early, but not late, block-
ade of the IL-10 receptor resulted in a significant, and sustained increase in antigen-specific antibody titers
during cognate iNKT cell help, but had no effect when traditional CD4+ T cell help was present. We con-
clude that the early composite cytokine environment is critical for dictating the long-term course of the B
cell response. Based on these data, we suggest that B cell antigens which recruit only cognate help from
iNKT cells experience a regulatory rather than inflammatory environment.
Poster 17
47.
SLAMf6 Modulates the NKT Cell Death Threshold
Victoria L. DeVault1, Oliver Dienz1, Graham W.J. Lilley1, Patrick Benoit1, Pamela L. Schwartzberg2,
Jonathan E. Boyson1 1Department of Surgery, University of Vermont College of Medicine, Burlington, VT 2National Human Genome Research Institute (NHGRI), National Institutes of Health,
Bethesda, MD
Signaling lymphocyte activation marker family member 6 (SLAMf6) is a cell surface signaling re-
ceptor that plays a critical role in NKT cell development. Surprisingly, the exact mechanisms through
which SLAMf6 regulates NKT cell development and function remain unclear. To investigate the function
of SLAMf6 on peripheral NKT cells, we challenged C57BL/6 (B6) or B6.Slamf6-/- mice with the NKT cell
agonist, aGalCer. While we detected no difference between B6 and B6.Slamf6-/- mice in NKT cell IFN-g,
IL-4, and TNFa production, we did find a significant difference in NKT cell numbers in the periphery.
Three days after challenge, we observed a 50-fold increase in NKT cells in B6 mice over vehicle-treated
controls. In contrast, we found a 220-fold increase in NKT cells in B6.Slamf6-/- mice versus controls. A
comparison between B6 and B6.Slamf6-/- mice of in vivo BrdU uptake by NKT cells revealed no signifi-
cant differences in proliferation. We then compared NKT cell apoptosis using Annexin V staining and
live/dead discriminator 3 h after aGalCer administration. This analysis revealed a significantly lower per-
centage of apoptotic NKT cells in B6.Slamf6-/- versus B6 mice, suggesting that SLAMf6 expression on
NKT cells was associated with activation-induced cell death (AICD). Consistent with this observation, we
found significantly diminished expansion of sorted B6 NKT cells when they were stimulated by SLAMf6+
antigen-presenting cells in an in vitro cell culture system. We conclude that in the presence of a strong,
high-affinity agonist, SLAMf6 contributes to significantly increased NKT cell AICD and concomitant di-
minished NKT cell expansion. Interestingly, when we conducted similar comparisons in naïve mice under
homeostatic conditions, we observed an increased percentage of apoptotic NKT cells in B6.Slamf6-/- mice
versus their B6 counterparts, which was associated with lower NKT cell numbers in B6.Slamf6-/- mice.
Taken together, these data support a model in which SLAMf6 regulates NKT cell activation and death
thresholds depending on the strength of activation. These data also suggest that SLAMf6 blockade could
be a useful tool to manipulate NKT cell populations in vivo.
Poster 20
48.
Oral Poster Presentations
Session B
Tumor Immunology
Chairs: Drs. Michael Robek and
Yasmin Thanavala
49.
A Novel Barrier to Endogenous Anti-Tumor Immunity: Ovarian cancer
ascites-activated neutrophils suppress T cell proliferation in a contact-dependent mechanism
Kelly L. Singel1, ANM Nazmul H. Khan2, Kirsten B. Moysich3, Kunle Odunsi4,5, Brahm H. Segal1,2,6
Departments of 1Immunology, 2Medicine, 3Cancer Prevention and Control, 4Gynecologic Oncology, and
the 5Center for Immunotherapy, Roswell Park Cancer Institute, Buffalo, NY 6Department of Medicine, University at Buffalo Jacobs School of Medicine and Biomedical Sciences,
Buffalo, NY
Neutrophils are the first responders to infection and injury, and critical for antimicrobial host de-
fense. Through the generation of reactive oxidants, activation of granular constituents, and neutrophil ex-
tracellular traps (NETs), neutrophils target microbes and prevent their dissemination. While these path-
ways are beneficial in the context of trauma and infection, their roles in the context of tumor are less un-
derstood. Ovarian cancer (OC) is often diagnosed at advanced stages and presents with ascites. Necrosis is
a hallmark of advanced cancer and releases DAMPs that activate innate immune responses. Cytotoxic T
lymphocyte (CTL) immunity is critical in OC, and barriers to durable anti-tumor immunity include TAMs,
MDSCs, and Tregs. While activated neutrophils can kill tumor cells, knowledge is limited on the role of
activated neutrophils in the tumor microenvironment.
Our prior studies showed that granulocytic cells from ascites of patients with newly diagnosed OC
variably suppressed stimulated normal donor T cell proliferation ex vivo. In ascites, ~90% of cells were
inflammatory (CD45+) with varying proportions of neutrophils, monocytes, and lymphocytes. Neutrophils
comprised ~15% of CD45+ cells and the neutrophil:CD8+ T cell ratio was 1.5:1. In ex vivo studies, cell-
free ascites (CFA) attracted normal donor neutrophils (NDN) and induced NETs. We next evaluated the
effects of CFA-treated NDN on T cell proliferation. In co-culture studies, a subset of CFA (5/10 tested)
activated NDN to completely suppress CD3/CD28-stimulated T cell proliferation. Neither CFA nor NDN
alone impaired T cell proliferation as measured by [3H] thymidine at a 1:1 target:effector cell ratio. Sup-
pression did not affect T cell viability or induce apoptosis, but required T cell contact with NDN in the
presence of CFA. In addition, we found that PD-1 expression on the suppressed T cells phenocopied the
unstimulated controls. When T cells were CD3/CD28-stimulated overnight before exposure to CFA and
NDN, T cell proliferation was not inhibited by the addition of CFA and NDN, suggesting that CFA-treated
NDN interrupt T cell proliferation at an early stage of activation.
These results support a model in which neutrophils in the ascites of a subset of OC patients sup-
press T cell responses, which may be an important barrier to endogenous anti-tumor immunity and immu-
notherapy. Further studies will identify ascites constituents that activate neutrophils and mechanisms for
neutrophil-mediated T cell suppression. This work may lead to novel prognostic biomarkers regarding in-
flammatory responses in the OC tumor microenvironment and therapeutic approaches that target neutro-
phils.
Supported by the NIH (R01, CA188900; T32, CA085183), the RPCI-UPCI Ovarian Cancer SPORE (P50
CA159981), and the Cancer Center Support Grant to RPCI (CA016056).
Poster 4
50.
Tumor-induced Myeloid-derived Suppressor Cells Act via Remote Control to Inhibit L-selectin-
dependent Adaptive Immunity in Lymph Nodes
Amy W. Ku1, Jason B. Muhitch1, Colin A. Powers1, Michael Diehl1, Anand P. Sharda1,
Kieran O'Loughlin1, Hans Minderman1, Joseph J. Skitzki1, Suzanne Ostrand-Rosenberg2,
Scott I. Abrams1, and Sharon S. Evans1 1Roswell Park Cancer Institute, Buffalo, NY 14263, USA; 2University of Maryland Baltimore County,
Baltimore, MD 21250, USA
Myeloid derived suppressor cells (MDSC) are potent immunomodulatory cells that play an exten-
sive role in cancer progression and immune evasion. These immature myeloid cells are known to accumu-
late within the spleen and tumor, and their ability to suppress effector T cell functions within these tissues
is well described. In contrast, the impact of MDSC on naive T cells within lymph nodes (LN) has been
largely overlooked as MDSC are rare within these critical sites of immune priming. Previous reports have
shown that peripheral MDSC from tumor-bearing mice downregulate the LN homing receptor L-selectin
on naive CD4 and CD8 T lymphocytes, but the molecular mechanisms and target cell-specificity has been
unclear. Furthermore, the biological relevance of moderate fluctuations of L-selectin is questionable as the
high density of L-selectin molecules normally present on T cells could theoretically buffer against the ef-
fects of such loss during trafficking. Using stringent murine mammary tumor models of high and low
MDSC burden (4T1 and AT-3, respectively), we demonstrate that MDSC downregulate L-selectin on na-
ive T and B cells post-transcriptionally via a contact-dependent mechanism. MDSC-driven loss of L-
selectin occurs within 24 hours both in vitro and in vivo, and does not appear to be species-restricted as L-
selectin on human lymphocytes can also be targeted by MDSC. By employing real-time intravital micros-
copy and immunofluorescence histology to visualize and assess naive CD8 T cell trafficking within vascu-
lar gateways for lymphocyte trafficking known as high endothelial venules (HEV), we found that even
moderate losses of L-selectin mediated by MDSC causes a profound reduction in the quality of lympho-
cyte-HEV interactions. Ultimately, this results in significantly fewer T cells trafficking and infiltrating
into the LN parenchyma. In an in vivo vaccination model, MDSC-mediated loss of L-selectin on naive
CD8 T cell and subsequent reduction in lymphocyte trafficking severely diminishes antigen-driven T cell
expansion within draining LN. These data reveal a novel mechanism by which tumor-induced MDSC lo-
calized outside of the LN shape the magnitude of T cell responses within the intranodal compartment,
which has unanticipated implications for systemic immunity in cancer. Supported by the NIH (CA79765,
AI082039, T32CA085183, 5T32CA108456, 5P30 CA016056), the Breast Cancer Coalition of Rochester,
the Mark Diamond Research Fund and the Jennifer Linscott Tietgen Family Foundation.
Poster 24
51.
Tumor-Derived Indoleamine 2,3- Dioxygenase Regulates Density of Tumor
Infiltrating CD8+ T cells and Myeloid Derived Suppressor Cells in a Murine Model of Ovarian
Cancer
Adaobi Amobi1,3, and Kunle Odunsi1,2,3
Departments of Immunology1, Gynecologic Oncology2 and Center for Immunotherapy3
Roswell Park Cancer Institute, Buffalo, NY
Amino-acid withdrawal is an important molecular mechanism regulating anti-tumor immune re-
sponses. The catabolism of the essential amino-acid tryptophan (TRP) by indoleamine 2,3-dioxygenase
(IDO1) is a central pathway that contributes to the immunosuppressive microenvironment in many types
of cancer. IDO1 enzymatic activity results in depletion of TRP and the generation of immunosuppressive
metabolites, such as kynurenine. Our lab has previously shown that IDO1 expression in human ovarian
tumor correlates with poor prognosis and poor tumor infiltration by CD8+ T cells. Moreover, our lab
demonstrated that increased infiltration of CD8+ T cells into the tumor is associated with improved sur-
vival. Thus, IDO1 inhibition represents an attractive target for cancer immunotherapy.
To establish the mechanism by which IDO1 inhibition augments immune responses in a murine
model of metastatic ovarian cancer, we utilized a murine ovarian surface epithelial cancer cell line, IE9-
mp1. We generated a stable IDO1-overexpressing cell line (IE9mp1-mIDO1) by transfecting murine IDO
cDNA into parental IE9-mp1 cells and confirmed functional IDO1 enzyme activity. C57BL/6 mice were
challenged intraperitoneally with either parental IE9mp1-Empty Vector (IE9mp1-EV) or IE9mp1-
mIDO1 tumor cells. Syngeneic immunocompetent mice inoculated with IE9mp1-mIDO1 cells displayed
earlier onset of tumor burden and decreased overall survival compared with IE9mp1-EV challenged
mice.
To delineate the role of host- and tumor-derived IDO1 on immune cell infiltration to the tumor
site, we utilized the IDO1 genetic knockout (IDO1KO) mouse model. IDO1KO and C57BL/6 mice were
challenged intraperitoneally with either the IE9mp1-EV or IE9mp1-mIDO1 tumor cells. C57BL/6 and
IDO1KO mice challenged with IE9mp1-mIDO1 demonstrate reduced CD8+ T cell infiltration within the
tumor. Interestingly, IDO1KO mice challenged with IE9mp1-EV tumor cells demonstrate increased tu-
mor infiltration by CD8+ T cells compared to C57BL/6 mice. Moreover, tumor-derived IDO1 mediates
increased frequency in the CD11b+Gr1+ myeloid derived suppressor cell (MDSC) population in ascites
fluid early-on along tumor burden in C57BL/6 and IDO1KO tumor-bearing animals.
From these data, we conclude that regulation of IDO1 will promote anti-tumor immune responses,
by permitting increased frequency of effector T cells in tumor tissues. Moreover, tumor-derived IDO1
inhibition may decrease the frequency of CD11b+Gr1+ MDSCs in ascites fluid. Future studies are ongo-
ing to further delineate the specific contribution of IDO1 by tumor cells, host cells, or both mutually to
the regulation of immunosuppressive MDSCs in ovarian cancer. Experiments are ongoing to characterize
the mechanism by which IDO1 inhibition may augment vaccine-induced immune responses in a murine
model of ovarian cancer.
Supported by: NCI SPORE P50 CA159981
Poster 26
52.
Negative Impact of Myeloid-derived Suppressor Cells on CD8 Effector T cell
Trafficking Within the Tumor Microenvironment
Amy Ku1, Michelle Appenheimer1, Jason Muhitch2, Scott I. Abrams1, and Sharon S. Evans1
Departments of Immunology1 and Urology2, Roswell Park Cancer Institute, Buffalo, NY
The success of T cell-based immunotherapy and, unexpectedly, thermal therapy, standard chemo-
therapy and radiation hinges on cytotoxic T cells gaining access to tumor targets. These observations have
prompted interest in strategies to improve T cell trafficking to tumors although the mechanisms that posi-
tively or negatively regulate extravasation at tumor vascular checkpoints are poorly understood. Here, we
report that the ability of tumor vessels to respond to IL-6-dependent preconditioning regimens that boost
CD8 effector T cell homing is temporally and inversely related to the accumulation of myeloid-derived
suppressor cells (MDSC) within the tumor microenvironment. Using real-time intravital imaging and im-
munofluorescence histology, IL-6 therapies were shown to convert vessels from T cell-low to -high recruit-
ment sites in murine tumors with minimal MDSC infiltration (i.e., CT26 colorectal, B16 melanoma, EMT6
mammary tumors). This conversion requires induction of the ICAM-1 trafficking molecule on tumor ves-
sels. Conversely, mammary (4T1, AT-3 and PyMT-MMTV) and pancreatic (Pan02) tumors with high
MDSC burdens were refractory to IL-6 therapies, but became responsive after acute MDSC depletion. To
further investigate contributions of MDSC to poor trafficking, IL-6-responsive tumors were admixed with
syngeneic CD11b+Gr-1+ MDSC isolated from spleens of tumor-bearing mice at a ratio of 2:1, thus mim-
icking the high MDSC burden detected in IL-6-refractive tumors. Sustained intratumoral elevation of
MDSC in admixed tumors resulted in failure to support increased T cell trafficking in response to IL-6–
dependent therapies. Complementary in vitro studies revealed that MDSC directly influence and downreg-
ulate trafficking molecule expression on endothelial cells. Taken together, these findings identify a novel
role of MDSC in subverting antitumor immunity by limiting T cell trafficking at tumor vascular loci. Sup-
ported by NIH (R01CA79765, R01AI082039, 2T32CA085183), the Breast Cancer Coalition of Rochester,
the Mark Diamond Research Fund and the Jennifer Linscott Tietgen Family Foundation.
Poster 27
53.
Pretreatment Peripheral Blood Monocyte Subset Signature is Predictive of Patient Response to
Dendritic Cell Vaccination
Anand Sharda1, Alexander Wald1, Mohammad Habiby Kermany1, Katja Koeppen2, Thomas Hampton2,
Jan Fisher3, Camilo Fadul3, Marc Ernstoff4, Thomas Schwaab1, Jason Muhitch1,5
Department of Urology1, Medicine4, and Immunology5 Roswell Park Cancer Institute, Buffalo, NY De-
partments of Microbiology and Immunology2, and Medicine3 Geisel School of Medicine at Dartmouth,
Hanover, NH
Clinical trials have demonstrated that dendritic cell (DC) vaccination can initiate durable anti-
tumor immunity in a subset of cancer patients resulting in complete responses, even in stage IV Renal Cell
Carcinoma (RCC). The influence of monocytes, the starting material for conventional DC vaccines, on
patient responses remains under-investigated. Recently, three subsets of monocytes have been described
(classical, intermediate, and non-classical), each with distinct functional properties. However, their roles
in anti-tumor immunity, particularly in the context of DC vaccination, are unclear. The goal of this study
was to determine whether the circulating pretreatment monocyte subset gene expression and composition
from Stage IV RCC patients prior to DC vaccination predicted responses to treatment in a completed
phase II clinical trial (NCT00085436). Pretreatment circulating classical (CD14++, CD16-), intermediate
(CD14++, CD16+), and non-classical (CD14+, CD16++) monocyte subsets were isolated from patients. Pre-
treatment peripheral blood from complete responders (2 of 3 have no observable disease > 5 years follow-
ing therapy) contained fewer classical monocytes (57.5% ± 6.4) compared to all other groups (P < 0.05).
Interestingly, a higher percentage of DC derived from non-classical monocytes expressed costimulatory
molecules (CD80; 96.4%, CD86; 91.1%, HLA-DR; 99.1%) compared to classical monocyte-derived DC
(64.6%, 54.4%, 60.2%, respectively). DC derived from non-classical monocytes were also superior in
their ability to induce allogeneic T cell proliferation compared to DC originating from classical mono-
cytes. Additional gene expression analysis by unsupervised hierarchical clustering clearly distinguished
the transcriptional profile of classical, intermediate, and non-classical monocytes from RCC patients to
healthy controls. Further investigation revealed that monocytes from long term survivors (> 10 years)
could be distinctly segregated from other RCC patients. These findings demonstrate that DC-derived from
the minor CD16+ monocyte subset may represent a superior product for use in vaccination protocols. Gene
expression profiling of circulating monocytes may provide an accessible biomarker for patient responsive-
ness to immunotherapy. Future studies will address whether pretreatment levels of intermediate and non-
classical monocytes are prognostic indicators for response to additional immunotherapies, including
checkpoint blockade inhibitors.
Poster 48
54.
Patrick Murphy, Ph.D.
Technical Application Scientist
BioLegend
Flex-T™: Produce MHC Tetramers with Your Peptide of Interest
Identification of antigen-specific cytotoxic T-lymphocytes (CTLs) is important to understanding T
cell responses to infection and in both vaccine and immune therapy development. Production of MHC
class I tetramers however is a time-consuming, difficult process and remains a barrier to rapid identifica-
tion of antigen-specific CTLs. To simplify the generation of MHC class I tetramers, BioLegend has devel-
oped the Flex-T™ flexible MHC tetramer system. Flex-T™ is composed of MHC monomers loaded with a
UV-labile peptide that is degraded via use of a UV light source. When UV irradiation is performed in the
presence of the peptide of interest, peptide exchange occurs and the peptide of interest binds to the MHC I
peptide-binding groove. MHC monomers containing your peptide of interest are then made into tetramers
using fluor-conjugated streptavidin. In addition to being simple to use, Flex-T™ technology allows for
identification of up to 15 peptide targets per sample via combinatorial color coding. This advance reduces
the need for large volumes of blood to identify multiple rare CTL populations. Flex-T™ technology expe-
dites large-scale screening studies and allows for simple, cost-effective MHC class I tetramer production in
the lab.
55.
Oral Poster Presentations
Session C
Lymphocyte Immunobiology II
Chairs: Drs. Nicholas Mantis and
Gary Winslow
56.
Autoantigen Availability Determines the Innate Sensing Requirement During
Self-Antigen Driven Germinal Center Responses in Autoimmunity
Stephanie L. Schell, Chetna Soni, and Ziaur S.M. Rahman
Penn State College of Medicine, Hershey, PA
Systemic lupus erythematosus (SLE)-associated germinal center (GC) responses are driven by syn-
ergistic BCR, TLR, and IFN signaling. Aberrant selection processes within these GCs lead to the escape
of high-affinity, class-switched autoreactive B cells. The environmental factors that stimulate TLRs and
the regulation of TLR and IFN signaling events during autoreactive GC responses in vivo are incompletely
defined. Previous studies in our lab established that MerTK-deficient (Mer-/-) mice, which lack a critical
receptor expressed by macrophages and DCs that is involved in apoptotic cell clearance and TLR/IFN im-
munoregulatory signaling cascades, exhibit increased GC responses and T cell activation. Here, we used a
kinetic immunization-based approach in Mer-/- mice to drive apoptotic cell accumulation in GCs, allowing
us to evaluate the contribution of TLR-dependent self-ligand sensing and dampened immunoregulation on
GC formation and selection processes, under conditions of high and low autoantigen availability. As antic-
ipated from previous study of spontaneous GC responses in autoimmune-prone mice, TLR7-MyD88 sig-
naling significantly contributes to enhanced GC responses observed in Mer-/- mice, independent of autoan-
tigen load. Unexpectedly, there was also a kinetic dependence on TLR9 in autoantigen-driven GC re-
sponses, suggesting that self-DNA sensing by TLR9 may have a more complex and context-dependent role
in GC responses than initially postulated. Further, the deficiency of MyD88 under conditions of high auto-
antigen availability did not diminish GC response, potentially indicating a novel regulatory function for
MyD88 or its dependent pathways whereby the sensing of ligand present in cytosolic spillover caused by
high autoantigen load is inhibited. Mechanistically, Mer-deficiency promotes enhanced GC responses in
GC B cell-intrinsic and APC-dependent manners, with process/cell-type dependent requirements for TLR
signaling. As a result of altered immune activation, the loss of Mer compounded aberrant GC selection
and downstream kidney pathology in autoimmune-prone B6.Sle1b mice deficient for Mer (B6.Sle1b.Mer-/-
). Altogether, these results suggest that TLR signaling plays a complex role in self-antigen driven GC re-
sponses, whereby both kinetics and autoantigen load determine the requirement for different self-ligands
and the activation of their relevant receptors during response. Mer-deficiency also promotes autoimmunity
by synergizing with genetic susceptibility loci to dysregulate selection processes within GCs.
Poster 13
57.
IgM Memory B cells Reconstitute Multiple B Cell Lineages and Provide Protection
Kevin Kenderes, Amber Papillion, and Gary Winslow
SUNY Upstate Medical University
IgM memory B cells are now recognized as an important component of immunological memory.
They have been proposed to be a reservoir of broadly-reactive B cells that differentiate, in germinal cen-
ters, into high affinity class-switched B cells following antigen encounter. We provide evidence that a
highly-purified IgM memory B cell population can follow multiple pathways of differentiation after chal-
lenge infection, and demonstrate that the antibodies produced by these cells can provide protective immun-
ity. Our experimental model uses Ehrlichia muris, an intracellular tick-borne bacterium that generates a
robust CD11c+ T-bet+ IgM memory B cell population in C57BL/6 mice. Due to the presence of pre-
existing antibodies, investigation of the secondary IgM memory B cell response to ehrlichial infection had
not been possible. We are able to monitor EYFP-labeled IgM memory B cells after transfer of splenocytes
into naïve mice and observed differentiation EYFP-labeled cells into all effector and memory B cell line-
ages following secondary infection. This was accompanied by a 4-fold increase in IgM production, relative
to infected mice that did not receive memory cells. However, a small population of EYFP-labeled switched
memory cells was also found in the donor spleen cells. Therefore, to determined if IgM memory cells were
solely responsible for the reconstitution of the memory and effector B cell lineages we monitored highly
purified EYFP-labeled spleen IgM memory B cells following their transfer into naïve recipient mice. After
challenge infection, some donor memory B cells differentiated into IgM-producing plasmablasts and CD19
-negative plasma cells. Other donor B cells entered germinal centers, down-regulated CD11c, underwent
class switching, and generated switched memory B cells. Yet other donor cells were maintained as IgM
memory cells. Donor IgM memory B cells also protected the recipient mice from the fatal erhlichial infec-
tion, Ixodes ovatus ehrlichia (IOE), demonstrating the importance of IgM memory cells for protective sec-
ondary responses. Thus, during secondary responses, IgM memory cells can differentiate into IgM-
producing plasmablasts, switched germinal center cells, or switched memory cells, or undergo self-
renewal. These data reveal that IgM memory B cells are capable of generating protective secondary re-
sponses that can replenish many, if not all, effector and memory B cell lineages, thereby contributing to
long-term immunity to pathogens.
Poster 22
58.
CD28 Induces Metabolic Fitness in LLPCs through NFkB-Mediated Irf4 Expression and ROS-
Dependent Survival
Adam Utley, James Cooper, Louise Carlson, Peng Peng, Amin Mahpour, and Kelvin Lee.
Roswell Park Cancer Institute, Buffalo NY
Sustained humoral immunity is dependent upon the continual production of antigen-specific anti-
bodies by plasma cells. Upon activation, B cells differentiate into short-lived plasma cells (SLPCs) that
traffic to secondary lymphoid organs such as the spleen where they live for days to weeks then die by
apoptosis. In a second non-mutually exclusive model, B cells can differentiate into long-lived plasma cells
(LLPCs) that home to specialized survival niches in the bone marrow and live indefinitely. Much work
has gone into describing the competitive BM survival niche; however, the cellular and molecular interac-
tions which govern this survival program are incompletely understood.
We have published that CD28, the canonical T-cell costimulatory molecule, is required for LLPC
survival. In T cells CD28 is known to induce glycolysis at the expense of mitochondrial respiration. To
our great surprise, CD28 increased mitochondrial respiration in LLPCs whilst not affecting glycolysis di-
rectly. CD28 increased Glut1 expression and subsequent LLPC glucose uptake, as well as the glycolytic
capacity. This suggests that CD28 regulates the ability of LLPCs to successfully compete for nutrients in
the BM niche for long term survival and antibody production.
Mechanistically, CD28 induces NFkB-dependent IRf4 upregulation, known to regulate LLPC sur-
vival. Furthermore, inhibition of NFkB abrogates the CD28-induced increases in glucose uptake and mito-
chondrial mass. We recently published that the Grb2/Vav binding domain on the CD28 cytoplasmic tail is
required for LLPC survival. In mice wherein this domain is mutated (AYAA mutants), LLPCs have de-
creased Irf4 expression, glucose uptake and mitochondrial mass. This facilitates a model wherein CD28
induces NFkB dependent Irf4 through Grb2/Vav for metabolic fitness. We also describe an NFkB super-
enhancer element upstream of the Irf4, suggesting that CD28 may govern direct Irf4 promoter activity as
well as DNA folding. Irf4 knock down decreased PC mitochondrial mass, demonstrating that Irf4 may
directly regulate LLPC metabolic fitness.
One byproduct of mitochondrial respiration is the production of reactive oxygen species (ROS).
CD28 increased ROS specifically in LLPCs. Paradoxically, ROS inhibition prevented CD28-mediated
survival. Taken together these results suggest a model wherein CD28 through its Grb2/Vav binding do-
main induces NFkB dependent upregulation of Irf4 directly through the promoter region, and augments
further Irf4 production through a previously undescribed NFkB superenhancer element. Irf4 then goes on
to increase mitochondrial respiration-dependent ROS for CD28-mediated LLPC survival and metabolic
fitness. Targeting CD28 with FDA-approved drugs may augment vaccine design as well as alleviate anti-
body mediated auto-immunity.
Poster 23
59.
Commensal Microbes Drive the Generation of Systemic IgA Responses
Joel R. Wilmore, Brian Gaudette, Wenzhao Meng, Eline T. Luning Prak, and David Allman
Department of Pathology and Laboratory Medicine, Perelman School of Medicine at the University of
Pennsylvania, Philadelphia, PA
It is well known that IgA functions as a critical component of the mucosal barrier in the gut by vir-
tue of its ability to be secreted into the intestinal lumen. However, little is known about the origin, function
and regulation of IgA in the serum. Mucosal IgA responses have been generally thought of as short-lived
and restricted locally to mucosal tissues. However, we find that IgA-secreting PCs make up the majority of
the BM PC pool in standard C57BL/6 mice bred in our colony. The IgA+ BM PCs are predominantly in the
long-lived pool and express gut homing factors such as CCR9 and the integrin α4β7 (alpha4 beta7), sug-
gesting a mucosal origin. The extent to which the commensal microflora influences the BM plasma cell
pool is evident by mice bred in germ free isolators that lack IgA+ PCs in their BM. Additionally, mice
from standard vendors such as Jackson labs (Jax), have extremely low levels of IgA+ PCs in the BM and
significantly lower serum IgA. Exposing Jax mice to a disparate microflora or Helicobacter sp. led to the
generation of IgA-secreting BM cells, while also inducing increases in serum IgA antibodies enriched for
binding to several commensal bacterial taxa. Moreover, BM IgA-secreting plasma cells exhibited a com-
mon clonal ancestry with intestinal IgA+ plasma cells, and both populations possessed unique gene expres-
sion signatures compared to other long-lived BM plasma cells. We conclude that commensal microbes
overtly influence the BM plasma cell pool, and suggest that select commensal microbes can facilitate the
induction of systemic humoral immunity.
Poster 38
60.
Regulation of IgM Memory B cell Pool Size by the Inhibitory receptor FcRIIb
Amber Papillion and Gary Winslow
Upstate Medical University, Syracuse, NY
Ehrlichia muris infection generates a long-term CD11c/T-bet-positive IgM memory population in
the spleen (J. Immunol. 191:1240). Among the many surface markers that distinguish the IgM memory
cells from canonical B cells is the inhibitory Fc receptor, FcRIIb, which exhibited a 2-fold higher expres-
sion. We hypothesized FcRIIb negatively regulates IgM memory cells by binding immune complexes pre-
sent during low-level chronic infection. To investigate this question, we monitored the IgM memory cell
population in infected FcRIIb-deficient mice. Thirty days post-infection, the IgM memory B cells were
generated earlier, and were found at three-fold higher frequencies in FcRIIb-deficient mice, compared to
wild-type mice. This increase in the frequency of spleen IgM memory cells was due to an increase in cell
number, and was in turn associated with an increase in antigen-specific IgG. These data indicate that Fc
RIIb plays an important role in regulating the expansion and/or persistence of IgM memory cells in wild-
type mice under conditions where antigen-specific IgG is sufficient to control infection. Other studies re-
vealed that the IgM memory population in FcRIIb-deficient mice exhibited much lower expression of
FAS, CD40, BAFF-R, and TACI. We therefore proposed that FcRIIb signaling, likely via immune com-
plexes, acts in wild-type cells to regulate the size of the IgM memory cell pool, by maintaining the expres-
sion of FAS and other receptors that regulate cell survival. These data suggest a novel regulatory role for
FcRIIb in B cell memory.
Poster 44
61.
Oral Poster Presentations
Session D
Immunoregulation
Chairs: Drs. Eyal Amiel and
William O’Connor
62.
Cytokine-mediated Regulation of Oligodendrocyte Metabolism
Scott B. Minchenberg, Anthony F. Paredes, and Paul T. Massa
SUNY Upstate Medical University, Department of Microbiology & Immunology, Syracuse NY
Multiple Sclerosis (MS) is a debilitating autoimmune disease characterized by inflammatory-
mediated demyelination in the central nervous system. Our primary goal is to understand how inflammato-
ry processes can lead to oligodendrocyte dysfunction, axonal degeneration, and demyelination in the CNS.
Our lab discovered that the protein tyrosine phosphatase SHP-1, a major negative regulator of the immune
system, is expressed in oligodendrocytes and plays an essential role in regulating STAT1 and STAT3 acti-
vation in oligodendrocytes. Importantly, it has recently been discovered that both STAT1 and STAT3 are
important transcriptional regulators of metabolic genes. Metabolic regulation is particularly important for
oligodendrocytes because lipid synthesis is required for the production and maintenance of the myelin
sheath and energetic support for axons. Using freshly isolated O4+ oligodendrocytes, we determined that
SHP-1 deficient oligodendrocytes have significantly reduced glycolysis and oxidative phosphorylation
(OXPHOS) relative to oligodendrocytes of wild type mice. We were able to show that in vivo treatment
with IFN- (gamma), a potent activator of STAT1, is able to down regulate wild type oligodendrocyte me-
tabolism without diminishing oligodendrocyte viability. Further, when oligodendrocytes of SHP-1-
deficient mice were treated with IL-10, a potent STAT3 activator, we were able to rescue metabolic defi-
cits in SHP-1-deficient oligodendrocytes suggesting that SHP-1 is a major regulator of oligodendrocyte
metabolism via regulation of STAT1 and STAT3 activation. The understanding of how cytokines such as
IFN- (gamma) and IL-10 modulate metabolism in oligodendrocytes is extremely relevant in the context of
inflammatory mediated demyelinating diseases.
Poster 8
63.
Inflammation, Androgens and Macrophages in the Prostate: Are we missing the link?
Camila Rosat Consiglio1 and Sandra Gollnick1,2
Departments of Immunology1 and Cell Stress Biology2, Roswell Park Cancer Institute,
Buffalo, NY
Prostate cancer (PCa) has the highest malignancy incidence rates in men and is the second leading
cause of male cancer mortality. While the underlying causes of PCa still remain largely unknown, it is
known that androgens, inflammatory mediators and inflammatory cells, including macrophages, are im-
portant players in prostate tumorigenesis. Interestingly, it has been shown that androgen receptor (AR) ex-
pression by macrophages enhances prostatic intraepithelial neoplasia formation in PTEN+/- mice, suggest-
ing that macrophage promotion of tumorigenesis is linked to macrophage AR signaling. Although it is
known that androgens influence immunity, the consequences of AR signaling in macrophages are largely
unknown. Furthermore, the activity and regulation of macrophage AR in the context of prostate tumorigen-
esis is still unclear. To address this gap in the field, the current study aims on analyzing the role of AR in
macrophage homeostasis and in prostate tumorigenesis. Initial results have shown that macrophages ex-
press higher levels of AR protein when compared to monocytes. Polarization of macrophages with either
IFN-γ (gamma) or LPS led to an increase in AR levels. Interestingly, macrophages in G2/M phase of the
cell cycle express higher levels of AR than macrophages in G1 phase, indicating that not only is AR in-
volved in macrophage differentiation and M1 polarization, but also in macrophage cell cycle. Imagestream
analysis of macrophages indicated that AR has strong nuclear localization. In addition, we have confirmed
AR transcriptional activity in bone marrow-derived macrophages using a luciferase assay. Since our results
show that AR is involved in macrophage homeostasis, we next investigated its role in tumor-bearing ani-
mals. We observed an increase in AR levels in tumor-associated macrophages (TAMs) of TRAMP C2
prostate tumors when compared to macrophages from other tissues. This finding was also observed in a
head and neck cancer model (MTERL). This evidence points to a role of the tumor microenvironment in
modulating macrophage AR levels and potentially its activity. Since it is known that macrophage AR facil-
itates prostate tumorigenesis and that macrophage AR levels are higher in the tumor microenvironment, it
is possible that early stages of tumorigenesis could induce macrophage AR signaling, leading to a pro-
tumorigenic phenotype in these cells. Future aims for this project will focus on elucidating macrophage AR
transcriptional targets in homeostasis and in cancer, as well as characterizing macrophage AR function dur-
ing prostate tumorigenesis.
Poster 18
64.
Cell-Intrinsic Glycogen Metabolism Supports Early Activation and Maintains
Metabolite Homeostasis in Dendritic Cells
Phyu Thwe1, Angelo D’ Alessandro2, Princess Rodriguez1, and Eyal Amiel1,3
1University of Vermont, Cellular Molecular and Biomedical Science Program 2University of Colorado Denver
3University of Vermont, College of Nursing and Health Sciences.
As professional antigen presenting cells of the immune system, dendritic cells (DCs) serve as a
bridge between innate and adaptive immune responses. Activation of DCs by a stimulus through toll-like
receptors (TLRs) is coupled with an increase in metabolic demand that is fulfilled by a TLR-driven burst in
glycolytic reprogramming. Up-regulation of glycolysis in activated DCs provides metabolites required for
DC effector function, and inhibition of glycolysis impairs the post-activation survival and effector function
of these cells.
TLR-driven glycolysis is thought to be sustained primarily by increased glucose uptake via the in-
ducible glucose transporter 1 (GLUT1). However, whether glucose is sourced from extracellular or intra-
cellular stores during early glycolytic reprogramming in DCs is still not well-defined. We propose that cell
-intrinsic glycogen metabolism in DCs supports early glycolytic burst that is essential for TLR-driven acti-
vation. The functional importance of glycogen metabolism in the context of DC effector responses has not
been previously described. Our data indicate that glycogen metabolism supports the activation of DCs, par-
ticularly during early activation before the up-regulation of GLUT1 expression. We show that DCs express
the enzymes essential for glycogen metabolism and that glycogen metabolism is regulated upon TLR stim-
ulation. Inhibition of glycogen utilization in DCs impairs the expression of costimulatory molecules CD40
and CD86 in these cells. The ability of DCs to uptake antigens and stimulate T cells is also compromised
upon disruption of glycogen metabolism. In addition, our metabolomics data indicate that glycogen metab-
olism in DCs generates both glycolytic and TCA cycle intermediates and that glycogen-derived carbons
may support metabolic pathways distinct from free glucose catabolism. These data demonstrate that the
glycogen metabolism plays a significant role in metabolic homeostasis in DCs and define a novel metabol-
ic regulatory pathway that supports DC immune function.
Poster 21
65.
Macrophages Negatively Regulate Hematopoietic Stem Cells in Murine
Aplastic Anemia
Angelica Costello, Amanda McCabe, PhD, Julianne N.P. Smith, PhD, & Katherine C. MacNamara, PhD
Albany Medical College, Albany, NY
Aplastic anemia (AA) is a rare bone marrow (BM) failure syndrome characterized by T cell-
mediated bone marrow destruction and pancytopenia. AA can be genetic or acquired, with the latter caused
by such insults as radiation, chemicals, drugs, and infection. T cell-derived interferon-γ (IFN-γ) has been
implicated in driving disease, but the mechanisms of IFN-γ-mediated pathogenesis in hematopoietic failure
during AA are not well understood. We recently identified a role for IFN-γ signaling specifically in macro-
phages (Mϕs) in driving the transient loss of hematopoietic stem cells (HSCs) in murine ehrlichiosis. Here,
in a mouse model of AA, we find that BM-resident Mϕs are maintained in the BM despite the loss of other
hematopoietic cell types, and that this requires IFN-γ. Moreover, Mϕ depletion with clodronate liposomes
or abrogation of IFN-γ signaling in Mϕs during AA rescues the HSC pool and significantly improves sur-
vival. We observed similar numbers of T-bet+ T cells and comparable IFN-γ production in AA mice that
were depleted of Mϕs, suggesting that Mϕs are critical sensors of IFN-γ and drivers of disease during AA.
Despite this, we did not detect a significant reduction in inflammatory proteins when IFN-γ signaling is
abrogated or when Mϕs were depleted. The one notable exception, however, was the chemokine CCL5
(RANTES), which was highly expressed in the BM during AA and significantly reduced in mice depleted
of Mϕs or when Mϕs were unable to respond to IFN-γ. CCL5 production in BM fibroblastic cells is in-
duced via podoplanin (PDPN) signaling. In support of a role of PDPN signaling in driving increased CCL5
during AA, we detected a significant increase in PDPN+ BM-resident Mϕs during AA. Neutralization of
either CCL5 or PDPN during AA rescued HSC numbers. Moreover, anti-PDPN treatment rescued platelet
numbers and significantly improved survival. Altogether, we demonstrate a novel role for IFN-g in AA
pathogenesis whereby IFN-γ promotes increased PDPN+ Mϕs and enhanced production of CCL5, thus
driving HSC loss and thrombocytopenia.
Poster 40
66.
Antagonistic Control of Intestinal Wnt Expression by IBD-Associated Cytokines
Travis Walrath, Stephen Sharp M.D., Shanti D'Souza, Prabhu Tewari, and William O'Connor Jr, Ph.D.
Albany Medical College, Albany NY
Homeostasis of the intestinal epithelium depends on the complex interplay of a variety of mediators
including cytokines that control, among other processes, a gradient of Wnt proteins. Homeostasis can be
disrupted by acute and chronic inflammation such as Inflammatory Bowel Disease (IBD). During IBD a
high concentration of the TH1 and TH17 associated cytokines, interferon gamma (IFN-γ) and interleukin-
17A (IL-17A) respectively, is observed within the colonic lamina propria. IFN-γ has previously been
shown to be cytotoxic to the intestinal epithelium, in part through interference with canonical Wnt signal-
ing; this is associated with increased disease severity in IBD. In murine models of disease, IL-17A has
been shown to lessen the severity of induced IBD.
One important question in the field is how these cytokines govern the intestinal epithelial and adja-
cent lymphoid compartments to either amplify inflammation or support mucosal healing. Toward under-
standing how these cytokines regulate the colonic microenvironment, we undertook a study using murine
knockout strains, primary tissue treated ex vivo, and primary spheroid/organoid cultures treated with IFN-γ,
IL-17A, or a combination of both. We found that IFN-γ reduced canonical Wnt expression while inducing
expression of non-canonical Wnts in primary murine colon tissue. Additionally we found that this altered
balance of wnt expression is dependent on the presence of lymphocytes and further, that wnt5a is specifi-
cally induced in CD19+ cells in the colon. Interestingly, IL-17A was able to oppose the increased expres-
sion of wnt5a. Moreover, mice deficient in IL-17A exhibit enhanced colonic wnt5a and TH1 cell marker
expression in response to Citrobacter rodentium infection, suggesting that IL-17A is opposing infection-
induced wnt5a and the TH1 bias during active colonic inflammation.
Canonical wnts, such as Wnt3a, are necessary for maintaining the potency of stem cells in the in-
testinal stem cell niche and the proliferative capacity of the intestinal epithelium. Non-canonical wnts such
as Wnt5a, have been shown to inhibit canonical wnt signaling and support tissue damage during colitis by
supporting increased IFN-γ production. Therefore, our data suggest that IFN-γ promotes intestinal epitheli-
al dysfunction in part by promoting a shift from canonical to non-canonical Wnt expression and function in
the colon. Furthermore, we hypothesize that IL-17A interferes with the IFN- mediated shift in Wnt ex-
pression, and therefore may reduce IFN- mediated epithelial toxicity and facilitate mucosal healing.
Poster 49
67.
Workshop I
Wayne M. Yokoyama, M.D.
“Opportunities to Study the Immunology of Human Diseases”
Much is known about basic immunology from the study of mouse immune responses. But how do
we translate these basic concepts to the human immune response. I will present examples where the trans-
lation has been achieved in less than obvious ways, highlighting the challenges and opportunities to under-
stand human diseases.
68.
Arielle Ginsberg
Technical Application Specialist
BD Biosciences
“No Title or Abstract Provided”
69.
Symposium III
Cancer Immunity
Chair: Dr. Edith Lord
70.
Exploring the Mechanisms of Neurotrophin-mediated Immune Tolerance and Their Implications
for Autoimmunity and Cancer
Joseph Barbi1, Paolo D. A. Vignali2, Hong Yu2, Fan Pan2, Drew Pardoll2 1Roswell Park Cancer Institute, Department of Immunology, Buffalo, NY 2Johns Hopkins
University School of Medicine, Department of Oncology, Baltimore, MD
Regulatory T cells (Tregs) enforce immune homeostasis and self-tolerance and inhib-
it both natural and induced anti-tumor immunity. Consequently, there is considerable inter-
est in therapeutic Treg enhancement or antagonism to treat inflammatory/autoimmune dis-
eases and cancer, respectively. Targeting Tregs requires a comprehensive understanding of
the factors important for their maintenance, differentiation and function. Here we report that
neuritin, a conserved, gpi-anchored molecule important for the development, survival and
function of neurons, is highly expressed by induced and natural Tregs. Neuritin expression
was also found to promote the maintenance and function of the Treg pool. Additionally,
neuritin ablation undercut Treg suppression in vivo, and tumors grew poorly in in the ab-
sence of neuritin, which permitted the mobilization of more robust anti-tumor immunity.
This was marked by enhanced pro-inflammatory cytokine production by tumor-associated
leukocytes and reduced expression of the checkpoint inhibitor PD-1. Importantly, we also
found that in Tregs, neuritin expression was closely linked to the functional differentiation
of Tregs into an activated, peripheral tissue-homing phenotype. As such, neuritin deficiency
resulted in an imbalance between “central”- and “effector”-like Treg populations and func-
tions. A role for neuritin in the modulation of co-stimulation pathways in favor of tolero-
genic outcomes was also identified. These findings characterize this neurotrophin as a hith-
erto unappreciated immunoregulatory molecule and a potential target for therapies aimed at
the fine-tuning of Treg function in cancer and inflammatory/autoimmune diseases.
71.
In situ analysis of multiple immune cells in tumor and microenvironment
Fiona Ginty, Ph.D.
GE Global Research, Niskayuna, NY 12309
Tumor immune response is highly complex, involving several distinct cell lineages,
cell-cell interactions and regulation. Variable cell density, infiltration and functional status
can determine tumor progression and immunotherapy response. Routine IHC methodolo-
gies are limited in their ability to quantify cellular diversity, and genomic methods may not
account for spatial location or cell activation status. Multiplexed analysis methods provide a
way to combine multiple markers and distinguish/quantify different cell types, and poten-
tially decipher immunotherapy response mechanisms. This presentation will discuss novel
approaches for multiplexed analysis of immune cell markers, quantifying cell type and
number in the tumor and microenvironment.
72.
Symposium IV
Immune Mediated Disease
Chair: Dr. Wayne Yokoyama
73.
Reactive Oxygen Species Induce Virus-independent MAVS-oligomerization in
Systemic Lupus Erythematosus
Andreas Koenig1, Theresa Montgomery1, Michael Murphy2, Richard Hartley3, Ryan Kelly4,
Andras Perl4, Ralph Budd5, and Iwona A. Buskiewicz1*
Department of 1Pathology and 5Medicine, Vermont Center for Immunology and Infectious
Diseases, University of Vermont, Burlington, USA; 2MRC Mitochondrial Biology
Unit, Wellcome Trust/MRC, Cambridge, UK; 3WestCHEM School of Chemistry,
University of Glasgow, Glasgow, UK; 4Upstate University Hospital, State University of
New York, Syracuse, USA
Systemic lupus erythematosus (SLE) is a complex autoimmune disease associated
with multiple immunologic abnormalities, among which upregulation of type I interferon
(IFN-I) genes correlates strongly with disease activity. Here we show that mitochondrial an-
tiviral signaling protein (MAVS), which normally forms a complex with retinoic acid gene
I (RIG-I)-like helicases during viral infection, can be activated by oxidative stress
alone. We observe that MAVS oligomerization on the surface of the mitochondrial outer
membrane by oxidative stress leads to mitochondrial hyperpolarization, decreased ATP pro-
duction, and increased spare respiratory capacity. This virus-independent oligomerization of
MAVS also leads to the secretion of IFN-I and proinflammatory cytokines. Consistent with
this, inhibition of mitochondrial reactive oxygen species (ROS) by the mitochondria-targeted
antioxidant MitoQ prevents oligomerization of MAVS. Patients with SLE are known to mani-
fest hyperpolarized mitochondria and increased ROS in peripheral blood lymphocytes. Our
findings reveal that MAVS is spontaneously oligomerized in peripheral blood mononuclear
cells of SLE patients, but not in matched healthy control individuals. Furthermore, ROS-
mediated MAVS oligomerization and IFN-I production was greatly reduced in cells expressing
a MAVS-C79F variant that occurs in 30% of sub-Saharan Africans and has been linked with
reduced expression of IFN-I and milder SLE. Our findings suggest that spontaneous redox-
induced MAVS oligomerization in SLE patients contributes to the IFN-I signature characteris-
tic of this syndrome.
74.
Increasing Regulatory Tone to Suppress Allergic Inflammatory Responses
in the Lung
Timothy J Chapman PhD, Sara E Hillman, Sara A Knowlden PhD, Jason A Emo MS,
Steve N Georas MD
Division of Pulmonary and Critical Care Medicine, Department of Medicine, University of
Rochester Medical Center, Rochester NY
Allergic inflammation and asthma are the result of inflammatory immune responses
raised against otherwise innocuous inhaled allergens. To prevent these unwanted respons-
es, mucosal immune cells establish an anti-inflammatory state in the lung that must be over-
come in order to induce inflammation. We term this phenomenon ‘regulatory tone’. In a
mouse model of allergic inflammation, low-dose bacterial lipopolysaccharide (LPS) admin-
istered with ovalbumin (OVA) via inhalation to naïve mice resulted in Th2-type allergic in-
flammation. However, prior tolerizing exposure to OVA alone prevented subsequent LPS/
OVA-induced allergic inflammation. Raising the dose of adjuvant in tolerized mice result-
ed in breakdown of tolerance and allergic inflammation with a mixed Th2/Th17 phenotype.
Therefore, increasing regulatory tone in the lung can prevent sensitizing exposures to aller-
gens. However, allergen exposure in the context of more severe inflammation can result in
breakdown of tolerance and a qualitative difference in the resulting immune response.
These data suggest that the balance between regulatory and inflammatory cells in the lung
is a critical feature in determining the outcome of inhaled allergen exposure.
75.
Industry Panel
“Careers Outside of Academia”
Participants
Patrick Murphy, Ph.D. (BioLegend, Inc.)
Fiona Ginty, Ph.D. (GE Global)
Erik Puffer, Ph.D. (BD Biosciences)
Olivia Schneider, Ph.D. (Shenandoah)
76.
Workshop II
Thomas J. Braciale , M.D., Ph.D.
Carter Immunology Center, Department of Pathology and Molecular Medicine
University of Virginia Health Systems
“The Business End of Academic Research”
This workshop is aimed at junior investigators in the process of starting up (or contemplating the
startup) an independent biomedical research laboratory. We will track the process of starting up a laborato-
ry from reviewing an offer of employment letter, negotiating a startup package, to hiring a technician, and
recruiting graduate students and fellows. We will use as a starting point for discussion, a video by a junior
investigator recently appointed to an independent academic position, who will describe her thoughts and
impressions about the business of starting up a research laboratory.
77.
Workshop III
“Granstmanship, Funding, and Mock Peer Review”
NIH Members
Timothy Gondre-Lewis, Ph.D. (Program Officer/COR/ DAIT Training Officer)
Thomas J. Palker, Ph.D. (Program Officer)
B. Duane Price, Ph.D. (Senior Scientific Review Officer)
James Snyder, Ph.D. (Scientific Review Officer)
Mock Peer Review Panelists
Margaret Bynoe, Ph.D. (Cornell University)
Richard I. Enelow, M.D. (Dartmouth Medical College)
Jonathan A. Harton, Ph.D. (Albany Medical College)
Nicholas J. Mantis, Ph.D. (Wadsworth Center/NYSDOH)
78.
Symposium V
Immunity in Infectious Disease
Chair: Dr. Thomas Braciale
79.
Control of the CD8+ Effector Population in Influenza Infection
Richard I. Enelow, M.D.
Dartmouth Medical College
The controlled contraction of immune responses during the clearance of respiratory virus infection
is critical to resolution of inflammation and the limitation of lung injury. The kinetics of antiviral effector
CD8+ T cell contraction, and dampening of effector function, appears to be programmed during the initial
activation of naive T cells, and possibly prior. Among other negative regulators of immune responses, it
has long been appreciated that TNF-alpha signaling plays an important role in contraction of CD8+ effec-
tor T cells, though the precise mechanisms involved remain unclear. We've shown that the critical timing
of TNF-programmed contraction occurs within the first 24-48 hours after initial antigen recognition, and
plays little immunoregulatory role thereafter. Furthermore, the critical source of this early burst of TNF
production is the naive CD8+ T cell upon initial antigen recognition. We hypothesize that the ability of the
naive CD8+ T cell to produce an early TNF burst is critically dependent upon homeostatic type I interferon
signaling in the host milieu prior to infection. The IFN-mediated regulation of the early burst of TNF-a by
naive CD8+ T cells is quite complex, and appears dependent upon post-thymic peripheral T cell
"licensing", during which the impact of constitutive low-level IFN production confers the tendency to
mount this important early TNF response. Together theses activities have a direct impact on the kinetics of
the expansion and contraction of the T cell responses and effector activities during and after viral infection,
serving to limit lung injury and immunopathology.
80.
Mast Cells in Kaposi’s Sarcoma – A Model for the Pathogenesis of
KSHV-driven Oncogenesis
Arturo Barbachano-Guerrero and Christine A. King
Department of Microbiology and Immunology, SUNY Upstate Medical University,
Syracuse, NY, USA
Kaposi sarcoma (KS) is a multicentric, inflammatory-driven, angioproliferative tumor of endotheli-
al cell (EC) origin where KS-associated herpesvirus (KSHV) is the etiological agent. This “hemorrhagic
sarcoma” is characterized by, and dependent upon, KSHV viral infection and inflammatory mediators for
proliferation and survival. Sources for infectious KSHV and inflammatory factors required for initiation,
expansion, and maintenance of KSHV-induced KS lesions are not fully understood. Here we demonstrate
for the first time that mast cells (MCs), key innate immune cells with potent inflammatory and angiogenic
responses, are key components of KS lesions. In this study, we analyzed cutaneous, gut, lymph node and
lung lesions from patients with classic KS for the presence of MCs by IHC staining using the MC specific
anti-tryptase (a major mast cell granule product) and KSHV LANA-1 antibodies. We found that MCs
were present in all lesions, independent of location or stage and localized in large numbers to highly vascu-
lar areas. IHC clearly demonstrated a sub-population of MCs in lesions that were positive for KSHV LA-
NA. MCs exhibited extensive degranulation that resulted in the release of potent pre-formed pro-
inflammatory tryptase and histamine. MCs were fully permissive to KSHV infection in vitro with de novo
viral gene expression observed as early as 6h post-infection (p.i) and culminating in release of infectious
progeny capable of establishing latent infection in primary human endothelial cells by 24 h p.i.. Taken to-
gether, we demonstrate for the first time that human MCs latently infected with KSHV are associated with
KS in all locations and stages of lesions development, in vivo. This suggests a model where MCs accumu-
late at the vascular interface between healthy and malignant KS tissue, are a potential source of infectious
virus and also act as inflammatory drivers needed to maintain and advance lesions.
81.
Toxoplasma Infections of the Nervous System
Ira J. Blader
Department of Microbiology and Immunology
University at Buffalo
Toxoplasma gondii infections can be severe and even fatal after they reactivate in immunocompro-
mised individuals. In most cases, these patients develop toxoplasmic encephalitis who clinically present
with dizziness, headaches, and seizures. Why patients develop these symptoms is unknown and has been
the focus of work in our laboratory for the past several years. To address this question, we used a murine
model of toxoplasmic encephalitis and found that the mice develop spontaneous seizures that are long last-
ing and severe. Since seizures can develop due to decreased inhibitory neuronal synaptic activity, we in-
vestigated the structure and function of GABAergic synapses, which are the major inhibitory synapses in
the brain. We found that the localization of GAD67, which is the primary GABA biosynthetic enzyme in
the brain, changes from being clustered at synaptic termini to be diffusely localized throughout the axon
and synapse. We further find that Toxoplasma decreases host neuronal GABAergic signaling, which is
consistent with the hypothesis that GAD67 mislocalization reduces host GABA receptor signaling. Toxo-
plasma triggering of seizures and GAD67 mislocalization occurs in mice infected with type II but not type
III strain parasites indicating the involvement of a polymorphic parasite factor . Given that these strains
generate significant differences in inflammatory responses, we compared the activation state of microglia/
monocytes in the brains of these mice and observed that strains that triggered seizures also had more severe
microglial activation. Finally, we find that a microglia inhibitor reduces both seizures and GAD67 mislo-
calization. Taken together, these data indicate that differences in inflammatory responses within the brain
in Toxoplasma-infected mice lead to specific changes in host GABAergic signaling that likely underlie sei-
zures in toxoplasmosis patients.
82.
Keynote Speaker
Thomas J. Braciale, M.D., Ph.D. Director, Emeritus
Carter Immunology Center
Professor of Pathology and Molecular Medicine
University of Virginia
“Dendritic Cells in the Host Response to Stress or Dendritic Cells Can Do More Than Present Antigens”
Dendritic cells (DC) have long been appreciated as the predominant antigen presenting
cells for the induction of primary adaptive immune responses to many invading microorganisms.
In this capacity, DC represent important first responders to the "stress" of microbial infection.
This presentation will describe recent studies analyzing the role of distinct subsets of respiratory
DC in initiating and regulating the adaptive immune CD8+ T cell response to influenza A virus
infection. However microbial infection is only one of many types of stress encountered in the en-
vironment. Related studies will describe a novel role for DC in regulating stress erythropoiesis,
that is the production of erythrocytes following hypoxia and tissue inflammation. (Supported by
USPHS grants AI-15608 and HL-33391).
Dr. Braciale’s lab is interested in the host immune response to virus infection—specifically, studying the role of the
adaptive immune response in the clearance of both virus and virus-infected cells from the body with much of the
work focused on infection of the respiratory tract by Influenza Virus and Respiratory Syncytial Virus.
83.
2015 NYIC Poster Numbers
AAI Young Investigator Award and Oral Poster Presentation eBioscience Travel Award and Oral Poster Presentation
Poster No:
1 Safiehkhatoon Moshkani
2 Megan Peppenelli
3 Lauren M. Webb
4 Kelly L. Singel
5 Oyebola Oyesola
6 Weishan Huang
7 James Emo
8 Scott B. Minchenberg
9 Abhinit Nagar
10 Taylor J. Moon
11 Christina M. Post
12 Alicia Soucy
13 Stephanie L. Schell
14 Danielle Y.F. Twum
15 Praneet Kaur Sandhu
16 Tariq A. Bhat
17 Jennifer Yates
18 Camila Rosat Consiglio
19 Bhuvana Katkere
20 Victoria L. DeVault
21 Phyu Thwe
22 Kevin Kenderes
23 Adam Utley
24 Colin A. Powers
25 Olesea Cojohari
Poster No:
26 Adaobi Amobi
27 Amy Ku
28 Catherine G. Burke
29 Aditi Murthy
30 Kathy A. Green
31 Arturo Barbachano-Guerrero
32 Tiffany Coupet/Clair Palmer
33 Janell Veazey
34 Danielle E. Baranova
35 Donald Steiner
36 Chenyang Jiang
37 Sadikshya Bhandari
38 Joel R. Wilmore
39 Angelene F. Richards
40 Angelica Costello
41 Amanda Poon
42 Kirsten E. Dostie
43 Amy Thees
44 Amber Papillion
45 Jennifer Vella
46 Margaret L. Barlow
47 Shivana M. Maharaj
48 Anand Sharda
49 Travis Walrath
84.
Poster 1.
Highly Attenuated Vesicular Stomatitis Virus-Based Vectors as Therapeutic Vaccines for Chronic
Hepatitis B
Safiehkhatoon Moshkani1, Sabine Lang2, John K. Rose2, Michael D. Robek1 1Department of Immunology & Microbial Disease, Albany Medical College, Albany, NY
2Department of Pathology, Yale University, New Haven, CT
Despite the availability of an effective preventative vaccine, chronic hepatitis B virus (HBV) infec-
tion affects over 240 million people worldwide and causes serious diseases such as liver cirrhosis and
hepatocellular carcinoma. Current treatments for chronic hepatitis B typically control but do not eliminate
the infection. The immune response to HBV in chronically infected people is typically weak and ineffec-
tive; therefore, therapeutic vaccines that elicit a functional antiviral immune response may be an important
component of a treatment regimen that is capable of curing chronic HBV. Virus-based vectors are highly
immunogenic compared to non-replicating vaccine platforms, but safety concerns can limit their use. We
previously demonstrated that recombinant wild-type vesicular stomatitis virus (VSV) expressing the HBV
middle surface envelope protein (MHBs) elicits functional virus-specific T cell and antibody responses in
mouse models of acute and chronic HBV replication. However, VSV has some undesirable pathogenic
properties, and the use of this platform in humans will require further attenuation of the vector. We there-
fore generated a VSV-based vector that expresses MHBs and contains two attenuating mutations that syn-
ergistically reduce virus replication: translocation of the nucleocapsid gene to the fourth genome position
and truncation of the glycoprotein cytoplasmic tail. This vector was evaluated for immunogenicity, patho-
genesis, and anti-HBV function in mice. Compared to wild-type VSV, the highly attenuated VSV vector
displayed markedly reduced pathogenesis but induced similar MHBs-specific CD8+ T cell and antibody
responses. The memory CD8+ T cell responses elicited by the highly attenuated VSV vector prevented
HBV replication in mice that were subsequently challenged by transduction with adeno-associated virus
(AAV) encoding the HBV genome (AAV-HBV). In mice in which persistent HBV replication was first
established by AAV-HBV transduction, treatment with the highly attenuated VSV induced MHBs-specific
CD8+ T cell responses that corresponded with a reduction in serum and liver HBV antigens and nucleic
acids, respectively. The ability of VSV to induce a robust multi-specific T cell response capable of control-
ling HBV replication combined with the improved safety profile of the highly attenuated vector suggests
this platform offers a potential strategy for HBV therapeutic vaccination.
85.
Poster 2.
HCMV Mediated Signaling Induces The Synthesis Of Select Akt-Dependent Antiapoptotic Proteins
During Entry To Promote Survival Of Short-Lived Monocytes.
Megan Peppenelli (1), Kyle Arend (2), Olesea Cojohari (1), Nathaniel Moorman (2) and Gary Chan (1).
(1): SUNY Upstate Medical University, Syracuse, New York, United States
(2): University of North Carolina at Chapel Hill, Chapel Hill, North Carolina, United States
HCMV infection of immunocompromised individuals often leads to multi-system organ failure.
The development of multi-system organ failure is dependent on the ability of HCMV to spread to peripher-
al organs, which is mediated by blood monocytes. In order for monocytes to mediate spread, we have pre-
viously shown HCMV to extend the short 48-hour lifespan of monocytes. Mechanistically, HCMV upregu-
lated two specific cellular antiapoptotic proteins, myeloid leukemia sequence 1 (Mcl-1) and heat shock
protein 27 (HSP27), to block two proteolytic cleavages necessary for the formation of fully active caspase
3 and the subsequent initiation of apoptosis. We now show that HCMV more robustly upregulated Mcl-1
when compared to normal myeloid growth factors and that HCMV was the only myeloid survival factor to
rapidly induce HSP27 prior to the 48-h cell fate checkpoint. We determined that HCMV glycoproteins gB
and gH signal through the cellular epidermal growth factor receptor (EGFR) and αvβ3 integrin, respective-
ly, during viral entry in order to drive the increase of Mcl-1 and HSP27 in an Akt-dependent manner. Alt-
hough Akt is known to regulate protein stability and transcription, we found that gB- and gH-initiated sig-
naling preferentially and cooperatively stimulated the synthesis of Mcl-1 and HSP27 through mTOR-
mediated translation. Overall, these data suggest that the unique signaling network generated during the
viral entry process stimulates the upregulation of select antiapoptotic proteins allowing for the differentia-
tion of short-lived monocytes into long-lived macrophages, a key step in the viral dissemination strategy.
Contact: Gary Chan, [email protected]
86.
Poster 3.
Role of Notch signaling in basophils in Type 2 inflammation
Lauren M. Webb1, Rebecca L. Cubitt1, Everett Henry2, Mark C. Siracusa2 and Elia D. Tait Wojno1
1Baker Institute for Animal Health and Department of Microbiology and Immunology, College of Veteri-
nary Medicine, Cornell University, Ithaca, NY, USA. 2Center for Immunity and Inflammation, New Jersey
Medical School, Rutgers Biomedical and Health Sciences, Rutgers, The State University of New Jersey,
Newark, New Jersey, USA.
Basophils are a rare granulocyte population, making up just 1% of circulating immune cells. De-
spite their rarity, basophils expand and contribute to the development of host-protective type 2 inflamma-
tion and worm expulsion following infection with Trichuris muris, a murine model of human whipworm.
However, the pathways that regulate basophil expansion and function in this setting remain unclear. Previ-
ous work has shown that the Notch signaling pathway is important in lymphopoiesis and particularly in the
function of T helper 2 cells, but little is known about its role in innate cell function. Here, we have investi-
gated the importance of the Notch signaling pathway in controlling the function of basophils during Type 2
inflammation. We show for the first time that basophils selectively upregulate Notch under Type 2 inflam-
matory conditions. Further, while Notch is not required for the steady-state development of basophils, it is
required for their optimal cytokine production in response to such damage signals as IL-33. We are now
actively investigating how other facets of basophil function are affected by inhibition of Notch signaling
and are characterizing how the selective deletion of Notch signaling in basophils affects anti-helminth re-
sponses and worm expulsion during T. muris infection. These findings have wide reaching implications
both for our continued understanding of innate cell function in Type 2 immunity but also in the potential
development of therapeutics aimed at targeting Type 2 inflammation.
87.
Poster 4.
A Novel Barrier to Endogenous Anti-Tumor Immunity: Ovarian cancer ascites-activated
neutrophils suppress T cell proliferation in a contact-dependent mechanism
Kelly L. Singel1, ANM Nazmul H. Khan2, Kirsten B. Moysich3, Kunle Odunsi4,5, Brahm H. Segal1,2,6
Departments of 1Immunology, 2Medicine, 3Cancer Prevention and Control, 4Gynecologic Oncology, and
the 5Center for Immunotherapy, Roswell Park Cancer Institute, Buffalo, NY 6Department of Medicine, University at Buffalo Jacobs School of Medicine and Biomedical Sciences,
Buffalo, NY
Neutrophils are the first responders to infection and injury, and critical for antimicrobial host de-
fense. Through the generation of reactive oxidants, activation of granular constituents, and neutrophil ex-
tracellular traps (NETs), neutrophils target microbes and prevent their dissemination. While these path-
ways are beneficial in the context of trauma and infection, their roles in the context of tumor are less un-
derstood. Ovarian cancer (OC) is often diagnosed at advanced stages and presents with ascites. Necrosis is
a hallmark of advanced cancer and releases DAMPs that activate innate immune responses. Cytotoxic T
lymphocyte (CTL) immunity is critical in OC, and barriers to durable anti-tumor immunity include TAMs,
MDSCs, and Tregs. While activated neutrophils can kill tumor cells, knowledge is limited on the role of
activated neutrophils in the tumor microenvironment.
Our prior studies showed that granulocytic cells from ascites of patients with newly diagnosed OC
variably suppressed stimulated normal donor T cell proliferation ex vivo. In ascites, ~90% of cells were
inflammatory (CD45+) with varying proportions of neutrophils, monocytes, and lymphocytes. Neutrophils
comprised ~15% of CD45+ cells and the neutrophil:CD8+ T cell ratio was 1.5:1. In ex vivo studies, cell-free
ascites (CFA) attracted normal donor neutrophils (NDN) and induced NETs. We next evaluated the effects
of CFA-treated NDN on T cell proliferation. In co-culture studies, a subset of CFA (5/10 tested) activated
NDN to completely suppress CD3/CD28-stimulated T cell proliferation. Neither CFA nor NDN alone im-
paired T cell proliferation as measured by [3H] thymidine at a 1:1 target:effector cell ratio. Suppression did
not affect T cell viability or induce apoptosis, but required T cell contact with NDN in the presence of
CFA. In addition, we found that PD-1 expression on the suppressed T cells phenocopied the unstimulated
controls. When T cells were CD3/CD28-stimulated overnight before exposure to CFA and NDN, T cell
proliferation was not inhibited by the addition of CFA and NDN, suggesting that CFA-treated NDN inter-
rupt T cell proliferation at an early stage of activation.
These results support a model in which neutrophils in the ascites of a subset of OC patients sup-
press T cell responses, which may be an important barrier to endogenous anti-tumor immunity and immu-
notherapy. Further studies will identify ascites constituents that activate neutrophils and mechanisms for
neutrophil-mediated T cell suppression. This work may lead to novel prognostic biomarkers regarding in-
flammatory responses in the OC tumor microenvironment and therapeutic approaches that target neutro-
phils.
Supported by the NIH (R01, CA188900; T32, CA085183), the RPCI-UPCI Ovarian Cancer SPORE (P50
CA159981), and the Cancer Center Support Grant to RPCI (CA016056).
88.
Poster 5.
The prostaglandin D2 receptor CRTH2 mediates interleukin-33-elicited group 2 innate lymphoid cell
accumulation in tissues
Oyebola Oyesola1, Lauren M. Webb1, Rebecca Cubitt1, Elia Tait Wojno1 1 Baker Institute for Animal Health, College of Veterinary Medicine, Cornell University, Ithaca NY 14853,
United States
Group 2 innate lymphoid cells (ILC2s) are rare innate immune cells that contribute to the develop-
ment of type 2 inflammation. These cells and associated type 2 inflammation promote allergic lung in-
flammation, which affects nearly 8% of the general population in the United States. Previous studies have
shown that the accumulation of ILC2s at inflammatory sites is mediated by epithelial cell-derived cyto-
kines and alarmins, particularly interleukin-33 (IL-33). In addition, prostaglandin D2 (PGD2), a bioactive
lipid produced by mast cells, induces accumulation of ILC2s at inflammatory sites by binding to its recep-
tor chemoattractant receptor-homologous molecule expressed on Th2 cells (CRTH2), which is expressed
by ILC2s. However, whether the IL-33-IL-33 receptor (IL-33R) and PGD2-CRTH2 pathways coordinate or
intersect to regulate ILC2 responses during type 2 lung inflammation has not fully been explored in vivo.
In this study, we investigated the role of the PGD2-CRTH2 pathway during IL-33-elicited ILC2 accumula-
tion and type 2 inflammation in the lung. When wild-type mice were treated either systemically by intra-
peritoneal injection or locally by intranasal injection with recombinant murine IL-33 (rmIL-33) to induce
type 2 inflammation, ILC2 frequency and number increased in the lung parenchyma. In contrast, ILC2s
accumulated in the lung of mice deficient in CRTH2 (CRTH2KO) following intranasal but not intraperito-
neal treatment with rmIL-33. The defect in ILC2 accumulation in the CRTH2KO lung following systemic
rmIL-33 treatment was not due to differences in expression of the IL-33R, cell death, apoptosis, or cell
proliferation. Therefore, our data suggest that the PGD2-CRTH2 pathway acts downstream of IL-33 to par-
tially mediate ILC2 accumulation in the lung during type 2 inflammation that originates in the periphery,
acting via pathways other than those controlling cell responsiveness to IL-33, death, or proliferation. Fu-
ture studies will focus on determining if ILC2 accumulation in the lung in response to IL-33 is mediated
via CRTH2-dependent migration to or retention in the lung. A better understanding of how cytokines and
bioactive lipid mediators interact to regulate ILC2 responses will be important in informing the use and
development of drugs that target these pathways to treat ILC2-associated type 2 airway inflammation.
89.
Poster 6.
ITK signaling via IRF4 regulates the development and function of type 1 regulatory T cells
Weishan Huang*, Sabrina Solouki, Nicholas Koylass, and Avery August*
Department of Microbiology and Immunology, Cornell University, Ithaca, NY 14853.
Type 1 regulatory T (Tr1) cells lack the expression of Foxp3 but have significant regulatory func-
tion in suppressing inflammation and promoting tolerance, in part via their expression of the immunosup-
pressive cytokine IL-10. Tr1 cells differentiate in response to signals engaging T cell receptor (TCR) and/
or the regulatory cytokine milieu. The non-receptor tyrosine kinase ITK is a key modulator downstream of
TCR, playing critical roles in T cell development and function. Using mouse models carrying Foxp3RFP
and IL-10GFP dual reporters, we found that, in the absence of ITK, TCR activation-driven development of
Foxp3- IL-10+ Tr1 cells is severely impaired in various organs (spleen, blood, lung, gut, and fat). Itk-/-
mice were also deficient in mounting a mucosal Tr1 cell response during parasitic (Nippostrongylus Brasil-
iensis) and viral (Influenza A) infections.
Naïve Itk-/- thymic and splenic Foxp3- CD4+ T cells also exhibited severe deficiency in Tr1 differ-
entiation under Tr1 polarizing condition. Although Itk-/- CD4+ T cells proliferated under Tr1 differentiating
conditions, they failed to up-regulate IL-10, and Tr1 cell markers LAG3, CD49b, ICOS, PD-1, c-Maf,
AHR, and IRF4 to levels observed in WT cells, suggesting that ITK is critical for Tr1 cell fate program-
ming. Utilizing a transgenic mouse model carrying an allele sensitive mutant of ITK (ITKas) that allows
ITK kinase specific blockade by a small molecule 3MB-PP1, we determined that the expression of the
aforementioned markers, as well as the balance between IL-10 and IFN-g (gamma) production during Tr1
differentiation, are dependent on ITK kinase activity. Furthermore, using cells from an ITKas-Foxp3RFP/IL
-17AGFP dual reporter mouse model, we find that ITK kinase activity is required for optimal Th17 trans-
differentiation to Tr1 cells. We also find that inhibiting ITK kinase activity diminished Tr1 differentiation
by human CD4+ T cells. The requirement for ITK function during Tr1 cell development can be restored by
the expression of the transcription factor IRF4. Finally, specifically targeting ITK kinase activity in already
differentiated Tr1 cells diminished their suppressive function.
We conclude that the TCR/ITK/IRF4 pathway is required for the development and function of Tr1
cells, which may be targeted to modulate regulatory immunity for clinical benefit.
*Correspondence: Weishan Huang ([email protected]) and Avery August
90.
Poster 7.
Enrichment for TNF-α Producing Naïve T Cells in Premature Infants Is Not Secondary to Bystander
Activation from Clinical Chorioamnionitis
Jason Emo, Natalie Martinez, Heidie Hyuck, Gloria Pryhuber, Dave Topham, Kristin Scheible
University of Rochester School of Medicine and Dentistry, Rochester, NY, United States.
Background: Newborn T cells are predominantly naïve, but interestingly naïve T cells from preterm infants
have a higher capacity to make TNF-α than full term babies. In-utero infection (chorioamnionitis), a com-
mon condition causing premature delivery, exposes the fetus to inflammatory cytokines such as IL-6, IL-1-
β, IL-8, and TNF-α. These cytokines are known to promote bystander activation of CD4+T cells.
Hypothesis: We hypothesized that enhanced TNF-α-producing T cells observed in premature infants at
birth, is due to inflammatory cytokine conditions present in chorioamnionitis.
Objective: To identify plasma cytokine profiles at birth that associate with prematurity or chorioamnionitis,
and determine the relationship between these profiles and frequency of TNF-α naïve CD4+ T cells.
Design/Methods: Umbilical cord blood plasma and mononuclear cells (UCMC) from 26 chorio-exposed
and 26 non-exposed infants was collected from 32 preterm (<32 0/7 weeks gestational age) and 20 full
term (>37 0/7 weeks gestational age) infants. Multiplex assays, customized to detect T-cell activating and
secreted cytokines, were performed on plasma samples. Functional profiles of CD4+T cells in matched
isolated UCMC was performed using flow cytometry.
Results: CD4+CD45RA+TNF-α+ T cells trended higher in infants exposed to chorioamnionitis. Naïve
CD4+ T cells from preterm babies showed higher frequencies of TNF-α+ producers at lower gestational
ages (p<0.0001), which was still significant when controlling for chorioamnionitis. Principle Component
Analysis (PCA) of plasma cytokines separated subjects into several clusters. Prematurity associated with
PCA1, which represented the greatest (22%) variability. PCA5, associated with chorioamnionitis, but con-
tributed to only 7% of the variability. CD4+CD45RA+TNF-α+ T cells were significantly lower (p=0.05) at
birth in subjects that clustered with PCA1.
Conclusion: Our results do not support the hypothesis that enrichment for naïve TNF-α+ producing T cells
in preterm infant cord blood is a result of bystander activation from in utero infection. Naïve TNF-α-
producing T cells are highly correlated with younger gestational age, which suggests this may be a devel-
opmentally-related finding. Plasma cytokine profiles in preterm infants are associated with reduced TNF-
α+ T cells, and may be protective against propagation of inflammation by T cells.
91.
Poster 8.
Cytokine-mediated regulation of oligodendrocyte metabolism
Scott B. Minchenberg, Anthony F. Paredes, and Paul T. Massa
SUNY Upstate Medical University, Department of Microbiology & Immunology, Syracuse NY
Multiple Sclerosis (MS) is a debilitating autoimmune disease characterized by inflammatory-
mediated demyelination in the central nervous system. Our primary goal is to understand how inflammato-
ry processes can lead to oligodendrocyte dysfunction, axonal degeneration, and demyelination in the CNS.
Our lab discovered that the protein tyrosine phosphatase SHP-1, a major negative regulator of the immune
system, is expressed in oligodendrocytes and plays an essential role in regulating STAT1 and STAT3 acti-
vation in oligodendrocytes. Importantly, it has recently been discovered that both STAT1 and STAT3 are
important transcriptional regulators of metabolic genes. Metabolic regulation is particularly important for
oligodendrocytes because lipid synthesis is required for the production and maintenance of the myelin
sheath and energetic support for axons. Using freshly isolated O4+ oligodendrocytes, we determined that
SHP-1 deficient oligodendrocytes have significantly reduced glycolysis and oxidative phosphorylation
(OXPHOS) relative to oligodendrocytes of wild type mice. We were able to show that in vivo treatment
with IFN-g (gamma), a potent activator of STAT1, is able to down regulate wild type oligodendrocyte me-
tabolism without diminishing oligodendrocyte viability. Further, when oligodendrocytes of SHP-1-
deficient mice were treated with IL-10, a potent STAT3 activator, we were able to rescue metabolic defi-
cits in SHP-1-deficient oligodendrocytes suggesting that SHP-1 is a major regulator of oligodendrocyte
metabolism via regulation of STAT1 and STAT3 activation. The understanding of how cytokines such as
IFN-g (gamma) and IL-10 modulate metabolism in oligodendrocytes is extremely relevant in the context of
inflammatory mediated demyelinating diseases.
92.
Poster 9.
NLRP3 inflammasome activation is modulated by a non-Pyrin domain cysteine
Abhinit Nagar, Tabassum Rahman, Jonathan A. Harton, PhD
Department for Immunology and Microbial diseases
Albany Medical College, Albany, NY
NLRP3 inflammasome activation results in activation of caspase-1 that aids in eliciting inflamma-
tory response by cytokine maturation. While structurally divergent agonists activate the NLRP3 inflam-
masome, the specific mechanism is still unresolved. The crystal structure of NLRP3 highlights the pres-
ence of a disulfide bond between conserved cysteines 8 and 108 in and adjacent to the N-terminal pyrin
domain (PYD) which may be important for NLRP3:ASC interaction and NLRP3 processing of IL-1β
(Beta). Many NLRP3 agonists trigger production of reactive oxygen species (ROS) which has been specu-
lated to drive the C8-108 disulfide bond. C8 is highly conserved across species. We explored the contribu-
tion of these cysteines to the NLRP3 inflammasome by evaluating multiple cysteine substitution mutants.
We evaluated formation of a perinuclear, oligomeric NLRP3:ASC complex (the inflammasome ‘speck’) by
Time Of Flight Inflammasome Evaluation (TOFIE) and NLRP3 processing of IL-1β by inflammasome re-
constitution. Here, we provide evidence that C8 and C108 are not important for NLRP3:ASC interaction.
However, NLRP3 activation of IL-1β processing is regulated by C108. These results highlight potential
inflammasome regulatory role for C108 adjacent to NLRP3 PYD and suggest that the spacer sequence con-
taining C108 likely comprises an uncharacterized regulatory domain of NLRP3. Further, as C108 appear to
differentially regulate the inflammasome response to distinct agonists, these results discount the hypothesis
that NLRP3 inflammasome agonists share a unified activation mechanism.
93.
Poster 10.
P2Y2 Regulation of CXCR4 Function
Taylor J. Moon and Michael R. Elliott
David H. Smith Center for Vaccine Biology & Immunology
University of Rochester, Rochester, NY
CXCR4 is a G-protein-coupled chemokine receptor with critical roles in immune cell development,
survival, and tissue homing. CXCR4 is also one of two major co-receptors for HIV-1 infection of human
cells. Therefore, a better understanding of the molecular regulation of CXCR4 expression and function is
important for developing therapeutic strategies to target this receptor as a means for altering immune cell
motility and HIV-1 infection. Here we demonstrate a novel role for the purinergic receptor P2Y2 in regu-
lating CXCR4 signaling in lymphocytes and myeloid cells. P2Y2 is a G-protein-coupled receptor respon-
sive to extracellular nucleotides including ATP and UTP, and P2Y2 signaling has recently been reported to
enhance HIV-1 infection of CD4+ T cells. Our work shows that human Jurkat T cells stably expressing
P2Y2 have decreased CXCR4-dependent migration, and concurrently human THP-1 monocyte cells with
siRNA knockdown of P2Y2 exhibit increased CXCR4 function. P2Y2 regulation of migration is specific
to CXCR4, because migration to other chemoattractants is not affected by over expression or knock down
of P2Y2. Additionally, after testing HIV-1 infection of human Jurkat T cells, we found that P2Y2 expres-
sion also significantly decreases infection over time compared to P2Y2 null controls. Interestingly, mouse
P2Y2 does not inhibit HIV-1 infection, nor does a truncated form of human P2Y2 lacking the cytoplasmic
tail (aa 1-310). From these data, we hypothesize that human P2Y2 signaling negatively regulates the ex-
pression and function of CXCR4 at the surface of lymphocytes. Elucidating the specific mechanisms re-
sponsible for P2Y2 regulation of CXCR4 will help determine whether P2Y2 is an attractive target for in-
hibiting HIV-1 entry as well as for therapeutic strategies aimed at modulating immune cell mobilization.
94.
Poster 11.
The ancestral environment shapes antiviral CD8+ T cell responses across generations
Christina M. Post, Jason Myers, and B. Paige Lawrence
Department of Environmental Medicine, University of Rochester School of Medicine & Dentistry,
Rochester NY
Recent studies have linked health fates of great-grandchildren to environmental exposures of their
great grandparents. However, few studies have considered whether ancestral exposures influence the im-
mune system across generations. Here we report novel findings regarding transgenerational transmission of
altered T cell responses resulting from maternal (F0) exposure to an environmentally relevant aryl hydro-
carbon receptor (AHR) ligand. The AHR is a transcriptional regulator that plays diverse roles in cellular
function, including modulating immune responses. AHR ligands comprise several classes of pollutants,
such as dioxins and PCBs, as well as molecules from foods and other sources. AHR-binding pollutants
cross the placenta and are excreted in breast milk. In animal models and human populations, early life ex-
posure to dioxins and PCBs is associated with persistent defects in the offspring’s immune function. Using
a mouse model, maternal exposure to the AHR ligand and pollutant 2,3,7,8-tetrachlorodibenzo-p-dioxin
(TCDD) results in a significantly reduced CD8+ T cell response to influenza A virus (IAV) in the adult off-
spring (F1), compared to the response of infected offspring of control-treated dams. Specifically, there are
significantly fewer cytotoxic T lymphocytes (CTL; CD44hiCD62Llo), virus-specific CD8+ T cells, and
CD8+ T cells that produce interferon gamma (IFNγ). Transcriptomic analyses using sorted CD8+ T cells
from F1 offspring of TCDD and control dams support new evidence that triggering AHR during develop-
ment changes programming of senescence or exhaustion regulatory pathways. Follow up studies show in-
creased expression of proteins associated with hindered T cell responses, such as CTLA-4 and KLRG1 on
CD8+ T cells. We next asked whether the diminished CD8+ T cell response in the F1 generation was ob-
served in the F3 generation. We detected fewer CTL and virus-specific CD8+ T cells in the TCDD F3 line-
age, as well as increased expression of CTLA-4 and KLRG1 compared to control F3 lineage following
IAV infection. These data indicate that F0 maternal exposure to AHR ligands is capable of disrupting im-
mune function not only via direct activation of the AHR in the F1 generation, but also by reprogramming
immune responses in subsequent generations. This has broad implications for understanding how the envi-
ronment of prior generations shapes susceptibility to pathogens and antiviral immunity in later generations.
95.
Poster 12
Immune tolerance against pulmonary F. tularensis infection
Alicia Soucy and Dennis W. Metzger, Ph.D.
Department of Immunology and Microbial Disease, Albany Medical College, Albany, NY
F. tularensis, a CDC Tier 1 agent, is the causative agent of tularemia, and inhalation of only
10 CFU can result in fatal disease. Currently, there is no licensed vaccine against this pathogen; however,
it is known that vaccinating BALB/c mice with a less virulent F. tularensis holartica strain (LVS) can pro-
tect against pulmonary challenge with the highly virulent SchuS4 strain. Vaccination does not protect
C57Bl/6 mice. We hypothesize that, as a result of vaccination, BALB/c mice establish a strong TH17-
lik erecall immune response, which results in tolerance to the SchuS4 strain. To test this, we primed
and boosted mice intranasally with LVS. After three weeks, we challenged LVS-vaccinated mice with
SchuS4, and analyzed bacterial burden. BALB/c mice had a lower bacterial burden in their lungs compared
to C57Bl/6 mice at days 8 and 10. Interestingly, while BALB/c mice survived challenge, they still had a
high bacterial burden in their lungs three weeks after infection. We also analyzed bronchoalveolar lavage
fluid for cytokine secretion; BALB/c mice produced greater levels of IL-17 and IL-22, compared to unpro-
tected C57Bl/6 mice. As expected, we saw decreased survival of vaccinated IL-17R knockout mice follow-
ing SchuS4 infection. In contrast, C57Bl/6 mice expressed greater levels various pro‑inflammatory cyto-
kines, including IFNγ, IL-6, IL-1α, and IL-1β. Histological analysis showed that BALB/c mice had less
lung necrosis at day 8 following SchuS4 challenge compared to C57Bl/6 mice. BALB/c mice also had less
total protein in the bronchoalveolar lavage fluid by day 10 post-infection. These findings suggest that
BALB/c mice mount a strong IL-17 and IL-22 recall response compared to C57Bl/6 mice, which aids in
enhancing survival by limiting tissue damage, but not bacterial clearance. Future studies are being conduct-
ed to determine the precise mechanisms responsible for tolerance in BALB/c mice from pulmonary
SchuS4 infection. (Supported by NIH P01AI056320)
96.
Poster 13.
Autoantigen Availability Determines the Innate Sensing Requirement During Self-Antigen Driven
Germinal Center Responses in Autoimmunity
Stephanie L. Schell, Chetna Soni and Ziaur S.M. Rahman
Penn State College of Medicine, Hershey, PA
Systemic lupus erythematosus (SLE)-associated germinal center (GC) responses are driven by syn-
ergistic BCR, TLR, and IFN signaling. Aberrant selection processes within these GCs lead to the escape
of high-affinity, class-switched autoreactive B cells. The environmental factors that stimulate TLRs and
the regulation of TLR and IFN signaling events during autoreactive GC responses in vivo are incompletely
defined. Previous studies in our lab established that MerTK-deficient (Mer-/-) mice, which lack a critical
receptor expressed by macrophages and DCs that is involved in apoptotic cell clearance and TLR/IFN im-
munoregulatory signaling cascades, exhibit increased GC responses and T cell activation. Here, we used a
kinetic immunization-based approach in Mer-/- mice to drive apoptotic cell accumulation in GCs, allowing
us to evaluate the contribution of TLR-dependent self-ligand sensing and dampened immunoregulation on
GC formation and selection processes, under conditions of high and low autoantigen availability. As antic-
ipated from previous study of spontaneous GC responses in autoimmune-prone mice, TLR7-MyD88 sig-
naling significantly contributes to enhanced GC responses observed in Mer-/- mice, independent of autoan-
tigen load. Unexpectedly, there was also a kinetic dependence on TLR9 in autoantigen-driven GC re-
sponses, suggesting that self-DNA sensing by TLR9 may have a more complex and context-dependent role
in GC responses than initially postulated. Further, the deficiency of MyD88 under conditions of high auto-
antigen availability did not diminish GC response, potentially indicating a novel regulatory function for
MyD88 or its dependent pathways whereby the sensing of ligand present in cytosolic spillover caused by
high autoantigen load is inhibited. Mechanistically, Mer-deficiency promotes enhanced GC responses in
GC B cell-intrinsic and APC-dependent manners, with process/cell-type dependent requirements for TLR
signaling. As a result of altered immune activation, the loss of Mer compounded aberrant GC selection
and downstream kidney pathology in autoimmune-prone B6.Sle1b mice deficient for Mer (B6.Sle1b.Mer-/-
). Altogether, these results suggest that TLR signaling plays a complex role in self-antigen driven GC re-
sponses, whereby both kinetics and autoantigen load determine the requirement for different self-ligands
and the activation of their relevant receptors during response. Mer-deficiency also promotes autoimmunity
by synergizing with genetic susceptibility loci to dysregulate selection processes within GCs.
97.
Poster 14.
The myeloid transcription factor IRF8 is integral for macrophage-mediated control of tumor
metastasis
Danielle Y.F. Twum1, Michael Nemeth1, Austin Miller2 & Scott I. Abrams1 1Department of Immunology; 2Department of Biostatistics & Bioinformatics,
Roswell Park Cancer Institute, Elm and Carlton Streets, Buffalo, NY 14263
The ‘M1’ (immune stimulatory) or ‘M2’ (immune suppressive) macrophage response in neoplasia
is governed by transcriptional pathways likely regulated by stromal- or tumor-derived factors bathing the
tumor microenvironment. In neoplasia, notably breast cancer, a high infiltration of macrophages is associ-
ated with reduced overall survival as well as progression to metastasis, which is thought to involve an M1
to M2 shift that favors tumor growth. Although several transcriptional regulators that drive the M2 pheno-
type in the tumor microenvironment have been identified, transcription factors that drive or enforce the M1
phenotype are not as well defined. IRF8 (Interferon Regulatory Factor-8) is a transcriptional regulator of
multiple aspects of myeloid biology, including differentiation and functional status. In the case of func-
tion, it is well-known that IRF8 is critical for the expression of genes such as iNOS and IL-12p40, hall-
mark features of an M1 phenotype. However, the role of IRF8 in macrophage-tumor biology is unknown.
Based on this rationale, we hypothesized that IRF8 is important in governing the ‘M1 vs M2’ macrophage
response in malignancy, namely mammary cancer whereby tumor-associated macrophages have been
linked to disease progression. To test this hypothesis, we utilized an IRF8 knockout model (IRF8fl/fl)
whereby IRF8 is specifically deleted in the macrophage compartment (LysM-Cre) and compared the rate of
tumor growth, as well as propensity for lung metastasis. Importantly, using an implantable mouse mam-
mary tumor model, we observed an increased propensity of lung metastasis in IRF8-/- mice compared to the
appropriate counterpart controls. Altogether, these data indicate that IRF8 expression in macrophages can
impact tumor progression to metastasis, likely through altering the tumor-suppressing (M1) vs. tumor-
promoting (M2) phenotypes. Thus, IRF8 may represent a potentially novel therapeutic target to modulate
the macrophage phenotype in neoplastic settings, such as mammary cancer, whereby this myeloid response
is a critical determinant of outcome.
98.
Poster 15.
Differentiation of iPSC-derived T cells specific to HBV
Praneet Kaur Sandhu, Mohammad Haque, Kristin Fino, Jianxun (Jim) Song
Pennsylvania State University College of Medicine, Hershey PA
Hepatitis B virus (HBV) is a double stranded DNA virus that infects hepatocytes to cause acute or
chronic infection in humans. Approximately 240 million people suffer from chronic HBV infection, which
involves persistence of virus for more than 6 months in infected individuals and can result in development
of hepatocellular carcinoma (HCC) and liver cirrhosis. Current treatment of HBV involves administration
of nucleoside/nucleotide inhibitors and cytokines but these have various side effects and prone to escape
mutations by HBV. Cellular immunotherapy using T cells primed to target disease-specific antigen could
prove to be an effective strategy to target hepatocytes that are chronically infected with HBV. Induced plu-
ripotent stem cells (iPSCs) can serve as a source of T cells as they have the ability to differentiate into any
cell type within the body. The objective of this project is to develop HBV-specific T cells from iPSCs in
vitro, characterize the underlying in vitro T cell differentiation mechanisms and utilize these iPSC-derived,
HBV-specific T cells for therapy in a HBV mouse model.
HBV-specific TCR was cloned into dsRed containing retroviral vector and successfully used for
retroviral transduction into murine iPSCs to establish a stable HBV-TCR expressing iPSC cell line. HBV-
TCR iPSCs were co-cultured with OP9-DL1-DL4 cells, a stromal cell line expressing delta ligand 1 (DL1)
and delta ligand 4 (DL4) for Notch signaling, to induce T cell differentiation.
In vitro differentiation of iPSCs into T cells can be utilized as effective method for generating T
cells for immunotherapy. Moreover, the development of iPSC-derived, HBV-specific T cells would pave
way for a therapeutic intervention for HBV.
99.
Poster 16.
Smoke Exposure Impairs Immunity To Lung Infection
Tariq A. Bhat1, Amit A. Lugade1, Suresh G. Kalathil1, Paul N. Bogner2,Thomas H. Thatcher3, Richard P.
Phipps3, Patricia J. Sime3,4, and Yasmin M.Thanavala1 1Department of Immunology and 2Department of Pathology, Roswell Park Cancer Institute, Buffalo, NY,
USA, 3Department of Medicine and 4Department of Environmental Medicine, University of Rochester,
Rochester, NY 14620
Tobacco smoke is recognized to have a detrimental impact on public health. Despite aggressive ef-
forts to prevent smoking, approximately a billion individuals continue to smoke worldwide. Cigarette
smoke (CS) exposure is a key initiator of chronic pulmonary inflammation and a major risk factor in the
development of respiratory disorders including chronic obstructive pulmonary disease (COPD). Strikingly,
in developed nations secondhand smoke (SHS) exposure is the main risk factor for nonsmokers to develop
emphysema and COPD. While exposure to SHS is common, the magnitude of this problem is not fully ap-
preciated.
To understand the connections among tobacco smoke exposure, chronic pulmonary infection,
chronic inflammation, and changes in immunity, we developed a mouse model. A major goal of our studies
is to understand the increased susceptibility to lung infection following exposure to either mainstream CS
or SHS, and to evaluate therapies that can restore normal immune function.
Using this model, we have shown that CS exposure has a profound immunosuppressive effect on
the generation of innate and adaptive immune responses to chronic infection with nontypeable Haemophi-
lus influenzae (NTHI). Our studies provide an understanding of the mechanism by which prior and ongo-
ing CS exposure predisposes COPD patients to recurrent infections that lead to exacerbations and contrib-
ute to mortality.
Recognizing the detrimental effects of SHS on human health we also utilized our mouse model to
interrogate its impact on pulmonary inflammation, immunity to pulmonary infection and to vaccination.
We observed that prior chronic SHS exposure also induces pulmonary inflammation and significantly
dampens the development of innate and adaptive immunity against chronic infection. We further investi-
gated whether judicious therapeutic intervention with an anti-inflammatory and pro-resolving lipid media-
tor may help reverse the inflammatory damage caused by prior smoke exposure and showed that it is possi-
ble to mitigate the effects caused by SHS.
It is well accepted that lung inflammation caused by cigarette smoke exposure persists long-term.
We therefore examined if cessation of exposure to SHS alone or in combination with treatment can reduce
inflammatory changes in the lung and augment immune function.
By taking a multi-pronged approach we have clearly shown that prior exposure to either CS or SHS
worsens respiratory infection-mediated inflammatory milieu and leads to the generation of defective innate
and adaptive immune responses. Furthermore, our results demonstrate the dramatic beneficial effects of
cessation and therapy in limiting pulmonary injury and augmenting immune responses to infection and
vaccination. Our results have considerable clinical relevance.
These studies were supported by a FAMRI Clinical Innovators Award.
100.
Poster 17.
Early IL-10 signals favor regulatory B cell over memory B cell development during cognate iNKT cell help
Jennifer Yates1,4, Emilie Vomhof-Dekrey1, Paula Lanthier1, Katja Mohrs1, Thomas Hägglöf 2, Natacha Veerapen3, Gurdyal Besra3, Mikael Karlsson2, and Elizabeth Leadbetter1,5
1Trudeau Institute, Saranac Lake, NY 12983; 2Karolinska Institutet, Stockholm, Sweden; 3School of Bio-sciences University of Birmingham, Birmingham, UK; 4Wadsworth Center, NYSDOH, Albany, NY; 5Uni-versity of Texas Health Science Center at San Antonio, San Antonio, TX
Effective generation of humoral B cell memory is dependent upon help from CD4+ T cells. We and
others have found that invariant natural killer T (iNKT) cells can provide both cognate and non-cognate
helper signals to enhance B cell responses. While both cognate and non-cognate iNKT cell help induce
class-switched, antigen-specific humoral immune responses - only non-cognate iNKT cell help drives the
formation of humoral memory. Rather, cognate iNKT cell help drives an early, un-sustained expansion of
germinal center B cells and antigen-specific antibody production. Therefore, we posit that cognate help
provided to B cells by iNKT cells is fundamentally different from the help provided by conventional CD4+
T cells. We now find that glycolipid immunization drives considerable IL-10 transcription by many differ-
ent spleen cell populations including dendritic cells, plasmablasts, iNKT cells, and B regulatory cells. Cog-
nate iNKT cell help expands antigen-specific IL-10 producing B regulatory cells upon primary immuniza-
tion, and IL-10 producing iNKT10 cells following secondary antigen challenge. Early, but not late, block-
ade of the IL-10 receptor resulted in a significant, and sustained increase in antigen-specific antibody titers
during cognate iNKT cell help, but had no effect when traditional CD4+ T cell help was present. We con-
clude that the early composite cytokine environment is critical for dictating the long-term course of the B
cell response. Based on these data, we suggest that B cell antigens which recruit only cognate help from
iNKT cells experience a regulatory rather than inflammatory environment.
101.
Poster 18.
Inflammation, androgens and macrophages in the prostate: Are we missing the link?
Camila Rosat Consiglio1 and Sandra Gollnick1,2
Departments of Immunology1 and Cell Stress Biology2, Roswell Park Cancer Institute,
Buffalo, NY
Prostate cancer (PCa) has the highest malignancy incidence rates in men and is the second leading
cause of male cancer mortality. While the underlying causes of PCa still remain largely unknown, it is
known that androgens, inflammatory mediators and inflammatory cells, including macrophages, are im-
portant players in prostate tumorigenesis. Interestingly, it has been shown that androgen receptor (AR) ex-
pression by macrophages enhances prostatic intraepithelial neoplasia formation in PTEN+/- mice, suggest-
ing that macrophage promotion of tumorigenesis is linked to macrophage AR signaling. Although it is
known that androgens influence immunity, the consequences of AR signaling in macrophages are largely
unknown. Furthermore, the activity and regulation of macrophage AR in the context of prostate tumorigen-
esis is still unclear. To address this gap in the field, the current study aims on analyzing the role of AR in
macrophage homeostasis and in prostate tumorigenesis. Initial results have shown that macrophages ex-
press higher levels of AR protein when compared to monocytes. Polarization of macrophages with either
IFN-γ (gamma) or LPS led to an increase in AR levels. Interestingly, macrophages in G2/M phase of the
cell cycle express higher levels of AR than macrophages in G1 phase, indicating that not only is AR in-
volved in macrophage differentiation and M1 polarization, but also in macrophage cell cycle. Imagestream
analysis of macrophages indicated that AR has strong nuclear localization. In addition, we have confirmed
AR transcriptional activity in bone marrow-derived macrophages using a luciferase assay. Since our results
show that AR is involved in macrophage homeostasis, we next investigated its role in tumor-bearing ani-
mals. We observed an increase in AR levels in tumor-associated macrophages (TAMs) of TRAMP C2
prostate tumors when compared to macrophages from other tissues. This finding was also observed in a
head and neck cancer model (MTERL). This evidence points to a role of the tumor microenvironment in
modulating macrophage AR levels and potentially its activity. Since it is known that macrophage AR facil-
itates prostate tumorigenesis and that macrophage AR levels are higher in the tumor microenvironment, it
is possible that early stages of tumorigenesis could induce macrophage AR signaling, leading to a pro-
tumorigenic phenotype in these cells. Future aims for this project will focus on elucidating macrophage AR
transcriptional targets in homeostasis and in cancer, as well as characterizing macrophage AR function dur-
ing prostate tumorigenesis.
102.
Poster 19.
Selenoproteins regulate B cell functions by targeting B cell receptor-mediated antigen
presentation pathway
Bhuvana Katkere1, Rachel L. Markley1, David R. Williamson1, Ashley E. Shay1, Bradley A. Carlson2.
Kumble S. Prabhu1 and Girish S. Kirimanjeswara1
1The Department of Veterinary and Biomedical Sciences, Pennsylvania State University, University Park,
PA,16802
2Molecular Biology of Selenium Section, Mouse Cancer Genetics Program, National Cancer Institute, Na-
tional Institutes of Health, Bethesda, MD, 20892
The essential micronutrient selenium (Se) has been shown to influence the immune functions in
both animals and humans. A vast majority of biological effects of Se is mediated by selenoproteins, a class
of proteins that contain 21st amino acid selenocysteine. A few recent studies have revealed that selenopro-
teins modulate the functions of T cells and macrophages. However, few studies have investigated the di-
rect effect of Se or selenoproteins on B cell functions. We therefore sought to establish the role of Se and
selenoproteins in B cell activation and functions, which are dependent on B cell receptor (BCR) signaling.
Supplementation of B cell line A20uWT with Se as sodium selenite at 200 nmol concentration resulted in
increased rate of BCR endocytosis, calcium signaling, antigen trafficking to MIIC compartment, antigen
degradation and antigen presentation compared to the cells maintained on deficient (0 nmol) or adequate
levels (50 nmol) of sodium selenite. Similar results were observed when B cells were supplemented with
other selenium containing compounds that can be metabolized and incorporated as selenocysteine. Howev-
er, supplementation with selenium compounds that cannot be metabolized and incorporated as selenocyste-
ine did not alter BCR-mediated signaling suggesting that the BCR-mediated signaling is regulated by sele-
noproteins. These data were reproducible in primary B cells isolated from the spleens of mice fed with 0.4
ppm sodium selenite (supplemented) compared to the mice fed with 0.01 ppm (deficient) or 0.08 ppm
(adequate) sodium selenite. This phenomenon was specific to BCR as other cell surface receptors such as
transferrin receptor internalization was not affected by the Se status of cells. Further studies revealed that
Se supplemented B cells had significantly lower levels of reactive oxygen species (ROS) compared to defi-
cient and adequate cells. Pharmacological reduction of ROS in Se deficient and adequate B cells resulted in
increased rate of BCR endocytosis. In contrast, pharmacological induction of ROS in Se supplemented
cells significantly decreased the BCR endocytosis. Together, these data suggest that one or more seleno-
proteins regulate the redox status of B cells that in turn has effect on B cell antigen processing and presen-
tation. Our preliminary studies suggest that Glutathione peroxidase (Gpx) family of selenoproteins such as
Gpx1 may be directly involved in the regulation of BCR-mediated signaling. These data together suggest
that Se, via selenoproteins, regulate the B cell receptor-antigen trafficking and presentation to impact the
secondary immune responses providing novel insights into molecular mechanisms of immune regulation.
103.
Poster 20.
SLAMf6 modulates the NKT cell death threshold
Victoria L. DeVault1, Oliver Dienz1, Graham W.J. Lilley1, Patrick Benoit1,
Pamela L. Schwartzberg2, Jonathan E. Boyson1 1Department of Surgery, University of Vermont College of Medicine, Burlington, VT 2National Human Genome Research Institute (NHGRI), National Institutes of Health,
Bethesda, MD
Signaling lymphocyte activation marker family member 6 (SLAMf6) is a cell surface signaling
receptor that plays a critical role in NKT cell development. Surprisingly, the exact mechanisms through
which SLAMf6 regulates NKT cell development and function remain unclear. To investigate the function
of SLAMf6 on peripheral NKT cells, we challenged C57BL/6 (B6) or B6.Slamf6-/- mice with the NKT
cell agonist, aGalCer. While we detected no difference between B6 and B6.Slamf6-/- mice in NKT cell
IFN-g, IL-4, and TNFa production, we did find a significant difference in NKT cell numbers in the pe-
riphery. Three days after challenge, we observed a 50-fold increase in NKT cells in B6 mice over vehi-
cle-treated controls. In contrast, we found a 220-fold increase in NKT cells in B6.Slamf6-/- mice versus
controls. A comparison between B6 and B6.Slamf6-/- mice of in vivo BrdU uptake by NKT cells re-
vealed no significant differences in proliferation. We then compared NKT cell apoptosis using Annexin
V staining and live/dead discriminator 3 h after aGalCer administration. This analysis revealed a signifi-
cantly lower percentage of apoptotic NKT cells in B6.Slamf6-/- versus B6 mice, suggesting that SLAM-
f6 expression on NKT cells was associated with activation-induced cell death (AICD). Consistent with
this observation, we found significantly diminished expansion of sorted B6 NKT cells when they were
stimulated by SLAMf6+ antigen-presenting cells in an in vitro cell culture system. We conclude that in
the presence of a strong, high-affinity agonist, SLAMf6 contributes to significantly increased NKT cell
AICD and concomitant diminished NKT cell expansion. Interestingly, when we conducted similar com-
parisons in naïve mice under homeostatic conditions, we observed an increased percentage of apoptotic
NKT cells in B6.Slamf6-/- mice versus their B6 counterparts, which was associated with lower NKT cell
numbers in B6.Slamf6-/- mice. Taken together, these data support a model in which SLAMf6 regulates
NKT cell activation and death thresholds depending on the strength of activation. These data also suggest
that SLAMf6 blockade could be a useful tool to manipulate NKT cell populations in vivo.
104.
Poster 21.
Cell-Intrinsic Glycogen Metabolism Supports Early Activation and Maintains Metabolite
Homeostasis in Dendritic Cells
Phyu Thwe1, Angelo D’ Alessandro2, Princess Rodriguez1, and Eyal Amiel1,3
1. University of Vermont, Cellular Molecular and Biomedical Science Program
2. University of Colorado Denver
3. University of Vermont, College of Nursing and Health Sciences.
As professional antigen presenting cells of the immune system, dendritic cells (DCs) serve as a
bridge between innate and adaptive immune responses. Activation of DCs by a stimulus through toll-like
receptors (TLRs) is coupled with an increase in metabolic demand that is fulfilled by a TLR-driven burst in
glycolytic reprogramming. Up-regulation of glycolysis in activated DCs provides metabolites required for
DC effector function, and inhibition of glycolysis impairs the post-activation survival and effector function
of these cells.
TLR-driven glycolysis is thought to be sustained primarily by increased glucose uptake via the in-
ducible glucose transporter 1 (GLUT1). However, whether glucose is sourced from extracellular or intra-
cellular stores during early glycolytic reprogramming in DCs is still not well-defined. We propose that cell
-intrinsic glycogen metabolism in DCs supports early glycolytic burst that is essential for TLR-driven acti-
vation. The functional importance of glycogen metabolism in the context of DC effector responses has not
been previously described. Our data indicate that glycogen metabolism supports the activation of DCs, par-
ticularly during early activation before the up-regulation of GLUT1 expression. We show that DCs express
the enzymes essential for glycogen metabolism and that glycogen metabolism is regulated upon TLR stim-
ulation. Inhibition of glycogen utilization in DCs impairs the expression of costimulatory molecules CD40
and CD86 in these cells. The ability of DCs to uptake antigens and stimulate T cells is also compromised
upon disruption of glycogen metabolism. In addition, our metabolomics data indicate that glycogen metab-
olism in DCs generates both glycolytic and TCA cycle intermediates and that glycogen-derived carbons
may support metabolic pathways distinct from free glucose catabolism. These data demonstrate that the
glycogen metabolism plays a significant role in metabolic homeostasis in DCs and define a novel metabol-
ic regulatory pathway that supports DC immune function.
105.
Poster 22.
IgM memory B cells reconstitute multiple B cell lineages and provide protection
Kevin Kenderes, Amber Papillion, Gary Winslow
SUNY Upstate Medical University
IgM memory B cells are now recognized as an important component of immunological memory.
They have been proposed to be a reservoir of broadly-reactive B cells that differentiate, in germinal cen-
ters, into high affinity class-switched B cells following antigen encounter. We provide evidence that a
highly-purified IgM memory B cell population can follow multiple pathways of differentiation after chal-
lenge infection, and demonstrate that the antibodies produced by these cells can provide protective immun-
ity. Our experimental model uses Ehrlichia muris, an intracellular tick-borne bacterium that generates a
robust CD11c+ T-bet+ IgM memory B cell population in C57BL/6 mice. Due to the presence of pre-
existing antibodies, investigation of the secondary IgM memory B cell response to ehrlichial infection had
not been possible. We are able to monitor EYFP-labeled IgM memory B cells after transfer of splenocytes
into naïve mice and observed differentiation EYFP-labeled cells into all effector and memory B cell line-
ages following secondary infection. This was accompanied by a 4-fold increase in IgM production, relative
to infected mice that did not receive memory cells. However, a small population of EYFP-labeled switched
memory cells was also found in the donor spleen cells. Therefore, to determined if IgM memory cells were
solely responsible for the reconstitution of the memory and effector B cell lineages we monitored highly
purified EYFP-labeled spleen IgM memory B cells following their transfer into naïve recipient mice. After
challenge infection, some donor memory B cells differentiated into IgM-producing plasmablasts and CD19
-negative plasma cells. Other donor B cells entered germinal centers, down-regulated CD11c, underwent
class switching, and generated switched memory B cells. Yet other donor cells were maintained as IgM
memory cells. Donor IgM memory B cells also protected the recipient mice from the fatal erhlichial infec-
tion, Ixodes ovatus ehrlichia (IOE), demonstrating the importance of IgM memory cells for protective sec-
ondary responses. Thus, during secondary responses, IgM memory cells can differentiate into IgM-
producing plasmablasts, switched germinal center cells, or switched memory cells, or undergo self-
renewal. These data reveal that IgM memory B cells are capable of generating protective secondary re-
sponses that can replenish many, if not all, effector and memory B cell lineages, thereby contributing to
long-term immunity to pathogens.
106.
Poster 23.
CD28 Induces Metabolic Fitness in LLPCs through NFkB-Mediated Irf4 Expression and
ROS-Dependent Survival.
Adam Utley, James Cooper, Louise Carlson, Peng Peng, Amin Mahpour, and Kelvin Lee.
Roswell Park Cancer Institute, Buffalo NY
Sustained humoral immunity is dependent upon the continual production of antigen-specific anti-
bodies by plasma cells. Upon activation, B cells differentiate into short-lived plasma cells (SLPCs) that
traffic to secondary lymphoid organs such as the spleen where they live for days to weeks then die by
apoptosis. In a second non-mutually exclusive model, B cells can differentiate into long-lived plasma cells
(LLPCs) that home to specialized survival niches in the bone marrow and live indefinitely. Much work has
gone into describing the competitive BM survival niche; however, the cellular and molecular interactions
which govern this survival program are incompletely understood.
We have published that CD28, the canonical T-cell costimulatory molecule, is required for LLPC
survival. In T cells CD28 is known to induce glycolysis at the expense of mitochondrial respiration. To our
great surprise, CD28 increased mitochondrial respiration in LLPCs whilst not affecting glycolysis directly.
CD28 increased Glut1 expression and subsequent LLPC glucose uptake, as well as the glycolytic capacity.
This suggests that CD28 regulates the ability of LLPCs to successfully compete for nutrients in the BM
niche for long term survival and antibody production.
Mechanistically, CD28 induces NFkB-dependent IRf4 upregulation, known to regulate LLPC sur-
vival. Furthermore, inhibition of NFkB abrogates the CD28-induced increases in glucose uptake and mito-
chondrial mass. We recently published that the Grb2/Vav binding domain on the CD28 cytoplasmic tail is
required for LLPC survival. In mice wherein this domain is mutated (AYAA mutants), LLPCs have de-
creased Irf4 expression, glucose uptake and mitochondrial mass. This facilitates a model wherein CD28
induces NFkB dependent Irf4 through Grb2/Vav for metabolic fitness. We also describe an NFkB superen-
hancer element upstream of the Irf4, suggesting that CD28 may govern direct Irf4 promoter activity as well
as DNA folding. Irf4 knock down decreased PC mitochondrial mass, demonstrating that Irf4 may directly
regulate LLPC metabolic fitness.
One byproduct of mitochondrial respiration is the production of reactive oxygen species (ROS).
CD28 increased ROS specifically in LLPCs. Paradoxically, ROS inhibition prevented CD28-mediated sur-
vival. Taken together these results suggest a model wherein CD28 through its Grb2/Vav binding domain
induces NFkB dependent upregulation of Irf4 directly through the promoter region, and augments further
Irf4 production through a previously undescribed NFkB superenhancer element. Irf4 then goes on to in-
crease mitochondrial respiration-dependent ROS for CD28-mediated LLPC survival and metabolic fitness.
Targeting CD28 with FDA-approved drugs may augment vaccine design as well as alleviate antibody me-
diated auto-immunity.
107.
Poster 24.
Tumor-induced myeloid-derived suppressor cells act via remote control to inhibit L-selectin-
dependent adaptive immunity in lymph nodes
Amy W. Ku1, Jason B. Muhitch1, Colin A. Powers1, Michael Diehl1, Anand P. Sharda1,
Kieran O'Loughlin1, Hans Minderman1, Joseph J. Skitzki1, Suzanne Ostrand-Rosenberg2, Scott I. Abrams1,
Sharon S. Evans1
1Roswell Park Cancer Institute, Buffalo, NY 14263, USA; 2University of Maryland Baltimore County,
Baltimore, MD 21250, USA
Myeloid derived suppressor cells (MDSC) are potent immunomodulatory cells that play an exten-
sive role in cancer progression and immune evasion. These immature myeloid cells are known to accumu-
late within the spleen and tumor, and their ability to suppress effector T cell functions within these tissues
is well described. In contrast, the impact of MDSC on naive T cells within lymph nodes (LN) has been
largely overlooked as MDSC are rare within these critical sites of immune priming. Previous reports have
shown that peripheral MDSC from tumor-bearing mice downregulate the LN homing receptor L-selectin
on naive CD4 and CD8 T lymphocytes, but the molecular mechanisms and target cell-specificity has been
unclear. Furthermore, the biological relevance of moderate fluctuations of L-selectin is questionable as the
high density of L-selectin molecules normally present on T cells could theoretically buffer against the ef-
fects of such loss during trafficking. Using stringent murine mammary tumor models of high and low
MDSC burden (4T1 and AT-3, respectively), we demonstrate that MDSC downregulate L-selectin on na-
ive T and B cells post-transcriptionally via a contact-dependent mechanism. MDSC-driven loss of L-
selectin occurs within 24 hours both in vitro and in vivo, and does not appear to be species-restricted as L-
selectin on human lymphocytes can also be targeted by MDSC. By employing real-time intravital micros-
copy and immunofluorescence histology to visualize and assess naive CD8 T cell trafficking within vascu-
lar gateways for lymphocyte trafficking known as high endothelial venules (HEV), we found that even
moderate losses of L-selectin mediated by MDSC causes a profound reduction in the quality of lymphocyte
-HEV interactions. Ultimately, this results in significantly fewer T cells trafficking and infiltrating into the
LN parenchyma. In an in vivo vaccination model, MDSC-mediated loss of L-selectin on naive CD8 T cell
and subsequent reduction in lymphocyte trafficking severely diminishes antigen-driven T cell expansion
within draining LN. These data reveal a novel mechanism by which tumor-induced MDSC localized out-
side of the LN shape the magnitude of T cell responses within the intranodal compartment, which has un-
anticipated implications for systemic immunity in cancer. Supported by the NIH (CA79765, AI082039,
T32CA085183, 5T32CA108456, 5P30 CA016056), the Breast Cancer Coalition of Rochester, the Mark
Diamond Research Fund and the Jennifer Linscott Tietgen Family Foundation.
108.
Poster 25.
Small Molecule Inhibitors of the PI3K/SHIP-1/Akt Pathway Sensitize Human
Cytomegalovirus-infected Monocytes to Apoptosis
Olesea Cojohari, Megan A. Peppenelli, and Gary C. Chan
SUNY Upstate Medical University, Syracuse, NY
Blood monocytes are responsible for systemic human cytomegalovirus (HCMV) dissemination,
which can cause multiorgan failure in immunodeficient hosts, such as transplant recipients and patients un-
dergoing chemotherapy. Current anti-HCMV drugs target viral replication, rendering them ineffective in
monocytes where the virus does not replicate; thus, novel therapeutic targets are needed to eliminate
HCMV-infected monocytes and prevent viral spread. Monocytes are naturally short-lived cells. We previ-
ously showed HCMV manipulates monocytes to survive past their 48-hour life-span. Activation of Akt
(antiapoptotic protein) is critical for M-CSF (myeloid growth factor) induced monocyte survival. Howev-
er, Akt’s role in HCMV-infected monocytes is unknown. We now show that treatment with the Akt inhibi-
tor, MK-2206, resulted in infected-monocytes apoptosis, suggesting Akt is central to HCMV induction of
monocyte viability. Interestingly, Akt activation was enhanced and kinetically different with HCMV than
M-CSF, implying the virus uses a unique mechanism to regulate Akt. Using PI3K isoform inhibitors, we
show HCMV induces a switch from p110δ, the main PI3K isoform positively regulating Akt in leukocytes,
to p110β, to mediate Akt-dependent survival. We next examined SHIP-1, a negative Akt regulator, shown
to also mediate positive (non-canonical) Akt regulation in cancer cells. Treatment with a novel SHIP-1
inhibitor (3AC) annulled HCMV-induced Akt signaling and triggered apoptosis, suggesting HCMV ex-
ploits a two-pronged approach, through PI3K-p110β and SHIP-1, to stimulate infected-monocyte survival.
Finally, HCMV entry elicited phosphorylation-mediated inactivation of PTEN (another negative Akt regu-
lator), ensuring maximum Akt activity. Overall, our data indicate that targeting Akt, p110β, or SHIP-1
with small molecule inhibitors could be effective in eliminating HCMV-infected monocytes and prevent-
ing viral spread and pathogenesis.
109.
Poster 26.
Tumor-Derived Indoleamine 2,3- Dioxygenase Regulates Density of Tumor Infiltrating CD8+ T cells
and Myeloid Derived Suppressor Cells in a Murine Model of Ovarian Cancer
Adaobi Amobi1,3, and Kunle Odunsi1,2,3
Departments of Immunology1, Gynecologic Oncology2 and Center for Immunotherapy3
Roswell Park Cancer Institute, Buffalo, NY
Amino-acid withdrawal is an important molecular mechanism regulating anti-tumor immune re-
sponses. The catabolism of the essential amino-acid tryptophan (TRP) by indoleamine 2,3-dioxygenase
(IDO1) is a central pathway that contributes to the immunosuppressive microenvironment in many types of
cancer. IDO1 enzymatic activity results in depletion of TRP and the generation of immunosuppressive me-
tabolites, such as kynurenine. Our lab has previously shown that IDO1 expression in human ovarian tumor
correlates with poor prognosis and poor tumor infiltration by CD8+ T cells. Moreover, our lab demonstrat-
ed that increased infiltration of CD8+ T cells into the tumor is associated with improved survival. Thus,
IDO1 inhibition represents an attractive target for cancer immunotherapy.
To establish the mechanism by which IDO1 inhibition augments immune responses in a murine
model of metastatic ovarian cancer, we utilized a murine ovarian surface epithelial cancer cell line, IE9-
mp1. We generated a stable IDO1-overexpressing cell line (IE9mp1-mIDO1) by transfecting murine IDO
cDNA into parental IE9-mp1 cells and confirmed functional IDO1 enzyme activity. C57BL/6 mice were
challenged intraperitoneally with either parental IE9mp1-Empty Vector (IE9mp1-EV) or IE9mp1-mIDO1
tumor cells. Syngeneic immunocompetent mice inoculated with IE9mp1-mIDO1 cells displayed earlier
onset of tumor burden and decreased overall survival compared with IE9mp1-EV challenged mice.
To delineate the role of host- and tumor-derived IDO1 on immune cell infiltration to the tumor site,
we utilized the IDO1 genetic knockout (IDO1KO) mouse model. IDO1KO and C57BL/6 mice were chal-
lenged intraperitoneally with either the IE9mp1-EV or IE9mp1-mIDO1 tumor cells. C57BL/6 and
IDO1KO mice challenged with IE9mp1-mIDO1 demonstrate reduced CD8+ T cell infiltration within the
tumor. Interestingly, IDO1KO mice challenged with IE9mp1-EV tumor cells demonstrate increased tumor
infiltration by CD8+ T cells compared to C57BL/6 mice. Moreover, tumor-derived IDO1 mediates in-
creased frequency in the CD11b+Gr1+ myeloid derived suppressor cell (MDSC) population in ascites fluid
early-on along tumor burden in C57BL/6 and IDO1KO tumor-bearing animals.
From these data, we conclude that regulation of IDO1 will promote anti-tumor immune responses,
by permitting increased frequency of effector T cells in tumor tissues. Moreover, tumor-derived IDO1 inhi-
bition may decrease the frequency of CD11b+Gr1+ MDSCs in ascites fluid. Future studies are ongoing to
further delineate the specific contribution of IDO1 by tumor cells, host cells, or both mutually to the regu-
lation of immunosuppressive MDSCs in ovarian cancer. Experiments are ongoing to characterize the
mechanism by which IDO1 inhibition may augment vaccine-induced immune responses in a murine model
of ovarian cancer.
Supported by: NCI SPORE P50 CA159981
110.
Poster 27.
Negative impact of myeloid-derived suppressor cells on CD8 effector T cell trafficking within the tu-
mor microenvironment
Amy Ku1, Michelle Appenheimer1, Jason Muhitch2, Scott I. Abrams1, and Sharon S. Evans1
Departments of Immunology1 and Urology2, Roswell Park Cancer Institute, Buffalo, NY
The success of T cell-based immunotherapy and, unexpectedly, thermal therapy, standard chemo-
therapy and radiation hinges on cytotoxic T cells gaining access to tumor targets. These observations have
prompted interest in strategies to improve T cell trafficking to tumors although the mechanisms that posi-
tively or negatively regulate extravasation at tumor vascular checkpoints are poorly understood. Here, we
report that the ability of tumor vessels to respond to IL-6-dependent preconditioning regimens that boost
CD8 effector T cell homing is temporally and inversely related to the accumulation of myeloid-derived
suppressor cells (MDSC) within the tumor microenvironment. Using real-time intravital imaging and im-
munofluorescence histology, IL-6 therapies were shown to convert vessels from T cell-low to -high recruit-
ment sites in murine tumors with minimal MDSC infiltration (i.e., CT26 colorectal, B16 melanoma, EMT6
mammary tumors). This conversion requires induction of the ICAM-1 trafficking molecule on tumor ves-
sels. Conversely, mammary (4T1, AT-3 and PyMT-MMTV) and pancreatic (Pan02) tumors with high
MDSC burdens were refractory to IL-6 therapies, but became responsive after acute MDSC depletion. To
further investigate contributions of MDSC to poor trafficking, IL-6-responsive tumors were admixed with
syngeneic CD11b+Gr-1+ MDSC isolated from spleens of tumor-bearing mice at a ratio of 2:1, thus mimick-
ing the high MDSC burden detected in IL-6-refractive tumors. Sustained intratumoral elevation of MDSC
in admixed tumors resulted in failure to support increased T cell trafficking in response to IL-6–dependent
therapies. Complementary in vitro studies revealed that MDSC directly influence and downregulate traf-
ficking molecule expression on endothelial cells. Taken together, these findings identify a novel role of
MDSC in subverting antitumor immunity by limiting T cell trafficking at tumor vascular loci. Supported
by NIH (R01CA79765, R01AI082039, 2T32CA085183), the Breast Cancer Coalition of Rochester, the
Mark Diamond Research Fund and the Jennifer Linscott Tietgen Family Foundation.
111.
Poster 28.
Triggering the aryl hydrocarbon receptor during development durably programs
CD4+ T cell responses
Catherine G. Burke1, Lisbeth A. Boule2, Jason R. Myers3, and B. Paige Lawrence1,2 1Department of 1Microbiology and Immunology, 2Department of Environmental Medicine, and
3Genomics Research Center, University of Rochester Medical Center, Rochester, NY
Emerging evidence shows that early life environmental exposures lead to lasting changes in im-
mune function, including increased incidence and severity of respiratory infections. Among the environ-
mental factors associated with altered immune function are ligands of the aryl hydrocarbon receptor
(AHR). Yet, how inappropriate triggering of AHR during development changes immune responses later in
life remains poorly understood. Recently, we showed that developmental activation of the AHR using the
prototypical ligand 2,3,7,8-tetrachlorodibenzo-p- dioxin (TCDD) results in an impaired response to influ-
enza A virus infection (IAV) in adult offspring. Changes include significantly fewer conventional CD4+
T cell effector subsets in infected offspring of TCDD treated dams, compared to infected offspring of con-
trol-treated dams. Adoptive transfer experiments show that this poorer response reflects developmentally
induced events intrinsic to the CD4+ T cell lineage. The activation and differentiation of naïve CD4+ T
cells into functionally distinct conventional subsets involves the integration of multiple signaling path-
ways, providing many different genes and cellular processes that could be affected by AHR activation dur-
ing development. To address this, we used an unbiased transcriptomic approach to identify differentially
expressed genes (DEG) in naïve and activated CD4+ T cells from infected adult offspring of dams treated
with TCDD or control. Specifically, we sorted CD44lo (naïve) and CD44hi (activated) CD4+ T cells, and
used next generation RNA sequencing (RNA-seq). After alignment and mapping to the genome, we used
DESeq2 to identify DEGs, and Ingenuity Pathways Analysis (IPA) to identify cellular pathways changed
by developmental AHR activation. Developmental exposure altered several signaling pathways critical for
CD4+ T cell differentiation, cell cycle regulation checkpoints, and metabolism. Activated CD4+ T cells
had more DEGs compared to unactivated cells. In addition, many DEGs and pathways were unique to
CD44hi cells, indicating that some developmentally programmed changes in gene expression are cryptic
(i.e. only revealed after CD4+ T cells are activated). These results indicate that early life AHR activation
influences several pathways critical for CD4+ T cells to mount appropriate responses to immune challeng-
es later in life. As humans are regularly exposed to AHR ligands, this has broad spanning implications for
human health, and gives insight into putative mechanisms by which early life environmental exposures
result in long lasting deficits in immune function.
112.
Poster 29.
Tumor Hypoxia alters Anti-tumor Immune Responses: Implications for Radiotherapy
Aditi Murthy1, Scott A. Gerber1, Cameron J. Koch2, and Edith M. Lord1
1University of Rochester Medical Center, Rochester, NY 2 University of Pennsylvania, Perelman School of Medicine, Philadelphia, PA
Tumor hypoxia occurs due to the increase in demand for oxygen by the rapidly growing tumor cells
together with reduction in the supply of oxygen due to malformed and non-functional tumor vasculature.
Tumor hypoxia offers resistance to radiotherapy (RT) and chemotherapy. Interestingly, a new paradigm has
emerged suggesting that hypoxia may also suppress immunotherapy, however the mechanisms behind this
observation remain undetermined. Our laboratory and others have shown that IFN conditions the tumor
milieu and is important for the efficacy of RT. As a result, we hypothesized that hypoxia could inhibit IFN
mediated anti-tumor responses resulting in decreased RT efficacy. Hypoxia could be modulating the pro-
duction of intratumoral IFN, and/or reducing the ability of cells to respond to IFN. To test the first possi-
bility we utilized Colon-38, a murine colon adenocarcinoma tumor model, and measured intratumoral hy-
poxia by both flow cytometry and fluorescence microscopy using a monoclonal antibody that detects hy-
poxia-induced 2-nitroimidazole adducts from the drug EF5. We used this drug to demonstrate a time de-
pendent increase of hypoxia within untreated Colon-38 tumors. Unexpectedly, irradiation resulted in a de-
crease in total tumor hypoxia. We also detected various sub-populations of hypoxic immune cells in both
the untreated and irradiated groups, with macrophages being the majority population. Importantly, CD8+ T
cells, which are an important source of IFN and mediate effector anti-tumor responses, were present in
these tumors but were not co-localized with hypoxic regions. This deficiency of effector T cells in the hy-
poxic regions indicates that tumor cells in these areas may also escape from immunotherapy. To address the
second possibility of hypoxia induced inhibition we tested whether hypoxia could modulate the responsive-
ness to IFN in vitro on various cells that make up the tumor microenvironment, in particular tumor cells,
and immune cells. Indeed, we were able to demonstrate that hypoxia inhibited the induction of IFN-
stimulated genes in multiple mouse and human tumor cell lines as well as peripheral blood mononuclear
cells suggesting hypoxia could inhibit the responsiveness to IFN. We propose that tumors that are less hy-
poxic (e.g. after RT and immunotherapy) are more conducive to IFN and T cell responses, resulting in en-
hanced tumor control. Funded by NIH R01 CA28332 and AAI fellowship 058232-002.
113.
Poster 30
LP-BM5 retrovirus-augmented Monocytic Myeloid Derived Suppressor Cells, Checkpoint Regulator
VISTA, and miR-155, are all players in a profound murine Immunodeficiency-causing syndrome
Kathy A. Green 1, Li Wang1,2, Randolph J. Noelle1, and William R. Green1
1 The Geisel School of Medicine at Dartmouth, 2 Medical College of Wisconsin
Inhibition by myeloid derived suppressor cells (MDSC) against T-cell responses is well established
in tumor microenvironments. We demonstrated (Green et al., J. Virology, 2013) augmentation of mono-
cytic MDSCs (M-MDSCs) during infection of susceptible B6 mice by LP-BM5 retrovirus, which causes a
profound immunodeficiency. These M-MDSCs inhibited not only T-, but also the much less reported B-,
cell responsiveness in ex vivo suppression assays. The M-MDSC inhibition of stimulated T-cell prolifera-
tion and IFN-gamma production was almost completely iNOS/NO dependent, whereas M-MDSC suppres-
sion of B-cell responses was only ~50% iNOS/NO dependent.
In exploration of additional suppressive mechanism(s) in the unique M-MDSC inhibition of B-cell
responsiveness, we have recently reported that VISTA (Green et al., J. Virology, 2015), a newly described
negative checkpoint regulator, is involved in LP-BM5-augmented M-MDSC-mediated suppression of
stimulated ex vivo B-cell responses. We found that combining the use of reagents to block both iNOS/
NO and VISTA lead to an additive, if not synergistic, abrogation of M-MDSC suppression of B-cell re-
sponsiveness. In additional recent studies, in light of the reported significant involvement of miR-155 in B
-cell and myeloid cell function, we have focused on the use of M-MDSCs from 5-7 week LP-BM5 infected
miR-155 k.o. mice. Our results have consistently shown these M-MDSCs to be substantially less (30-
50%) suppressive compared to M-MDSCs from LP-BM5 infected Wt. mice. Also, at 10-12 weeks post
infection, miR-155 k.o. mice exhibit a selectively different kind of immunodeficiency compared to the dis-
ease of Wt. mice. Thus, spleen cells from miR-155 k.o. infected mice consistently present as significantly
more responsive to stimulation with B-cell polyclonal activators, such as anti-CD40 and IL-4, and have
significant less splenomegaly than LP-M5 infected Wt. mice. The infected spleens of 10-12 week miR-
155 k.o.s also contain substantially fewer M-MDSCs as compared to Wt. infected spleens. In an initial
experiment, involving adoptive transfer of M-MDSCs from donor LP-BM5 infected Wt. B6 to infected
miR-155 k.o. recipients, at 10 weeks post infection, we were able to significantly augment in the recipient
miR-155 k.o.: 1) splenomegaly, 2) splenic B-and T-cell immunodeficiency, and 3) the number of residing
splenic M-MDSCs, to levels almost comparable to those of Wt. infected mice.
These results highlight involvement of multiple and unique suppressive pathways in the under-
studied area of MDSC suppression of B-cell responses, and were compatible with a role for M-MDSC in
LP-BM5-induced immunodeficiency.
114.
Poster 31.
Dengue virus non-structural protein 1 activates mast cells and endothelial cells
Arturo Barbachano-Guerrero1, Timothy Endy2 and Christine A. King1 1Department of Microbiology and Immunology; 2Department of Medicine, Upstate Medical University,
Syracuse NY
Arboviruses are an emerging worldwide heath concern. Transmitted by the mosquito vector Aedes
ageypti, dengue virus, family Flaviviridae, is estimated to infect 390 million individuals annually and is
associated with enormous morbidity worldwide.
Dengue infection results in severe disease in a subset of patients characterized by pathological vascular
permeability and high levels of systemic inflammatory mediators. A small RNA virus, dengue encodes
ten proteins, of which non-structural protein one (NS1) is secreted as a hexamer during infection and re-
mains in circulation after the virus is cleared. The role of this highly conserved hexameric viral protein in
the pathogenesis of severe dengue disease is unknown.
Previous work in our laboratory and others has shown that pathogenesis of severe disease involves
activation of both innate immune mast cells (MCs) and endothelial cells (ECs). The location of MCs adja-
cent to and lining the endothelium allow for direct cross talk facilitating MC modulation of vascular he-
mostasis. Mast cells directly contribute to the systemic pro-inflammatory environment by release of large
amounts of inflammatory cytokines and chemokines in response to virus and more recent work demon-
strated modulation of EC permeability by NS1 directly.
We hypothesized that secreted NS1 promotes viral pathogenesis by directly activating MCs and
ECs leading to altered homeostasis and inflammation. To address our hypothesis we used a series of
complementary in vitro assays to asses MC and EC responses. Our data demonstrated NS1 induced acti-
vation of both MCs and ECs at multiple levels. At the transcriptional level, NS1 stimulation of ECs in-
duced the select upregulation of CCL5, E-selectin and IL-11, all markers of EC activation. Further-
more, western blot analysis of MC and ECs stimulated with increasing concentrations of NS1 demon-
strated a dose dependent increase in Akt activation at S473, a cellular hub of several regulatory and in-
flammatory pathways, including angiogenesis, vascular permeability and pro- inflammatory cytokine
production. To determine the downstream effect of NS1 activation on EC function we evaluated the
ability of NS1 to directly modulate EC intercellular organization via the ability to organize into complex
structures and to migrate. Using the tube formation and scratch assays we demonstrate NS1 potently
modifies by inducing tube formation and migration. Overall, our results demonstrate that dengue NS1
can induce MC and EC activation and potently modify basic EC functions. Together, our data suggest
that NS1 may contribute to the pathogenesis of dengue disease by directly key cellular components
linked to severe disease pathogenesis.
115.
Poster 32.
NKG2D ligand targeted Bispecific T Cell Engagers (BiTE) lead to robust antitumor activity against
diverse human tumors
Tiffany Coupet, Claire Palmer, and Charles L. Sentman
Center for Synthetic Immunity, Department of Microbiology & Immunology, Geisel School of Medicine
at Dartmouth, Lebanon, NH
Two new bispecific T cell engager (BiTE) molecules with specificity for NKG2D ligands were de-
veloped. One, designated huNKG2D-OKT3, was derived from the extracellular portion of the human
NKG2D receptor fused to a CD3ε (epsilon) binding single chain variable fragment (scFv). NKG2D is a
receptor expressed by both NK and T cells that has the ability to bind multiple ligands, including MICA,
expressed by a variety of malignant cells. A second molecule, B2-OKT3, was created in the tandem scFv
BiTE format that targets MICA on tumor cells and CD3 on T cells. The data show that these BiTEs specif-
ically activate T cells to kill several human tumor cell lines, representing both liquid and solid tumors. The
huNKG2D-OKT3 BITE had a lower affinity for recombinant MICA (rMICA) but induced greater T cell
cytotoxicity and cytokine production compared with B2-OKT3. Both BiTEs were able to trigger activated
human T cells and resting PBMCs. Culture of plate-bound rMICA with T cells and these BiTEs showed a
ligand density dependent production of IFNγ (gamma) by both CD4+ and CD8+ T cells. There was two-
fold more IFNγ (gamma) produced per CD8+ T cell and five-fold greater percentage of CD8+ T cells that
produced IFNγ (gamma) compared to CD4+ T cells. The data show robust anti-tumor activity using scFv
or receptor-based bispecific constructs and support their further development for clinical use against can-
cer.
116.
Poster 33.
Novel role for protein kinase D in regulating airway epithelial cytokine secretion and barrier
integrity
Janelle Veazey1, Tim Chapman2, Tim Smyth3, Sara Hillman2, Steve Georas1,2
Department of Microbiology and Immunology1, Department of Pulmonary and Critical Care
Medicine2, Department of Toxicology3, University of Rochester, Rochester NY
Background: Previous work from our lab identified the serine/threonine kinase protein kinase D (PKD)
as a key regulator of barrier integrity in virally-infected bronchial epithelial cells. Although PKD has been
implicated in various cellular functions including cell growth, motility, and cytokine secretion, little is
known about the expression or function of PKD in the airway.
Methods: Human bronchial epithelial cells (16HBE) cells were pre-treated with vehicle or 5 µM of the
competitive PKD inhibitor CRT0066101 (CRT) or 50 µM of the noncompetitive PKD inhibitor
CID755673 (CID) (Tocris). 2 hours later, cells were stimulated in vitro with or without the double strand-
ed RNA polyI:C (5 µg/ml). Cell supernatants were analyzed for IL-8 and IFN-lambda by ELISA (R&D
BioScience).
Wild-type 6-10 week old C57BL/6 mice were treated with either vehicle or CRT (100
µg) prior to challenge with 10 µg polyI:C. 24 hours later, bronchoalveolar lavage (BAL) fluids were col-
lected, and cell counts determined by hemacytometry and cytospin. BAL supernatants were analyzed by
ELISA for KC and IFN-lambda. PKD isoenzyme protein level in whole cell extracts of different organs
was analyzed by Western blot.
Results: Inhibiting PKD in 16HBE cells with 5 mM CRT prior to 5 µg/ml polyI:C treatment markedly
attenuated the production of both IL-8 and IFN-lambda. Using wild-type mice, we found that PKD inhi-
bition reduced polyI:C-induced KC levels and neutrophilia in BAL fluid across multiple doses of polyI:C
administered and in a dose-dependent manner with the PKD-inhibitor. The PKD inhibitor CRT also at-
tenuated polyI:C-induced IFN-lambda levels in BAL fluid. Finally, we determined that PKD3 protein
levels are more highly expressed in the lung, lymph node and spleen than PKD1, suggesting PKD3 is the
isoform driving cytokine release.
Conclusions: PKD3 may play a previously unsuspected role in regulating epithelial chemokine release
and neutrophil influx into the airway during respiratory viral infections. PKD3-mediated cytokine release
might also contribute to induction of an
anti-viral state, specifically via IFN-lambda regulation. Further study of the mechanism of PKD3-
mediated cytokine release in the airway epithelium may lead to new therapeutics in combating respirato-
ry pathogens.
117.
Poster 34.
Intestinal Immunity to Vibrio cholerae Involves Antibody-Mediated Effects on Multiple Bacterial
Cell Processes
Danielle E. Baranova, Kara J. Levinson, Nicholas J. Mantis
Wadsworth Center, New York State Department of Health, Albany NY; University at Albany, Albany NY
Secretory IgA, and to a lesser extent serum IgG, serves as a primary line of defense against enteric patho-
gens in the intestinal lumen, although exactly how antibodies protect against non-invasive bacteria like
Vibrio cholerae is not well understood. It is generally assumed that secretory antibodies function solely by
promoting cell-cell cross-linking, agglutination and subsequent clearance via peristalsis of antibody-
pathogen complexes from the intestinal lumen, a process referred to as immune exclusion. To better define
the immunological processes associated with intestinal immunity, we are investigating the interactions be-
tween V. cholerae and ZAC-3, a monoclonal antibody that recognizes a conserved surface exposed epitope
within lipopolysaccharide (LPS) coat. We have found that mAbs have direct effects on bacteria in vitro,
independent of antibody-mediated crosslinking. These effects include rapid motility arrest and the coinci-
dent appearance of surface blebs and deformations that are suggestive of outer membrane (OM) stress. We
have also shown that ZAC-3 treatment stalls V. cholerae motility in liquid and viscous environments. To-
gether this indicates that the bacterial response to secretory antibodies is multifaceted and goes beyond ag-
glutination. We now demonstrate by scanning electron microscopy that exposure of V. cholerae strain
O395 to ZAC-3 promotes the appearance of exopolysaccharide (EPS)-like extensions on the bacterial sur-
face. This EPS production is both time and dose dependent, as detected by a crystal violet assay. ZAC-3
Fab fragments did not induce EPS production, indicating that agglutination is necessary, but (from other
studies) not sufficient to trigger exopolysaccharide expression. While the exact composition of the EPS
remains unknown, we provide preliminary results that indicate that EPS production shields the bacterium
from complement-mediated killing, suggesting that V. cholerae “goes on the defensive” in response to
ZAC-3 treatment. Collectively, these results suggest a complex interplay between pathogen and secretory
antibodies in the intestinal lumen, which ultimately determines efficiency of infection.
118.
Poster 35.
Innate control of pneumonic Francisella tularensis infection through macrophage depletion
and polarization
Donald Steiner and Dennis W. Metzger
The bacterium Francisella tularensis is a Tier 1 select agent and the cause of tularemia. F. tularen-
sis is a facultative intracellular pathogen capable of replicating within the cytoplasm of macrophages, and
in the highly lethal respiratory form of tularemia, alveolar macrophages are the primary hosts in the early
stages of infection. We have found that depletion of alveolar macrophages protects mice against respirato-
ry tularemia induced using a low infectious dose of the virulent type A strain Schu S4. Mice protected in
this fashion do not develop a systemic inflammatory response, suggesting that macrophage depletion pre-
vents establishment of infection. Clodronate treatment followed by infection with antibody-opsonized F.
tularensis protects mice even against greater infectious doses. F. tularensis has been recovered from the
blood of protected mice 5 days after infection, but not from the spleens of the same mice 21 days after in-
fection, indicating that these mice develop a productive, systemic infection but ultimately clear the infec-
tion and survive. Neutrophil depletion abrogates this protection. This work represents evidence that im-
munologically naïve mice can be protected against type A F. tularensis infection through innate immunity.
Supported by NIH grant PO1 A1056320
119.
Poster 36.
House Dust Mites-induced Allergic Asthma in Sickle Cell Disease Mice
Chenyang Jiang and Steven M. Szczepanek
Department of Pathobiology and Veterinary Science, University of Connecticut, Storrs CT
House dust mites (HDM) are human allergens and the leading cause of allergic asthma in the US and
worldwide. Sickle cell disease (SCD), as the most common monogenic blood disorder, is accompanied by
massive co-morbid diseases, including allergic asthma. Asthma confers a higher rate of mortality in indi-
viduals with SCD, especially in children. Some studies demonstrated it that co-morbid allergic airway dis-
ease (AAD) in transgenic SCD mice is more severe than in wild-type (WT) mice when they are exposed to
ovalbumin (OVA) and aluminum hydroxide (alum), which is characterized by increasingly intense vaso-
occlusion and pulmonary inflammation. Given this information, we questioned if SCD mice exhibit as
fierce of co-morbid AAD when they are exposed to HDM extract, as do OVA/alum sensitized SCD mice.
To study this, SCD mice were intranasally inoculated with HDM for acute (5 weeks) and chronic (11
weeks) periods. Unexpectedly, when SCD mice were instilled with a low dose (1 ug (microgram)) of
HDM for an acute time period, the amount of total white blood cells per ml of bronchoalveolar lavage flu-
id (BALF) was similar to the amount found in WT mice. BALF leukocyte differential counting was not
statistically different between SCD mice and WT mice, with eosinophils approximating 23% and macro-
phages 77% of cells. Consistent with this, inflammation scoring for histologic slides of lungs from SCD
mice was not significantly different with scores from WT mice. After an acute exposure of high dose
HDM (25 ug(microgram)), the amount of total white blood cells per ml BALF, BALF leukocyte differen-
tial counting and inflammation scoring for histologic slides of lungs from SCD mice were same as those
from WT mice. In this case, eosinophils were the majority at approximately 81% instead of macrophages
at 17%. As with acute disease, chronic exposure of SCD and WT mice to a high dose of HDM resulted in
statistically similar results for all parameters. Acute inflammation turned into immune tolerance, as eosino-
phils decreased to approximately 40% and macrophages increased to 52% of BALF leukocytes in both
groups. These data indicate that HDM exposure causes similar AAD in SCD mice and WT mice, which is
in stark contrast to the reports of severe AAD in SCD mice when they are run through the OVA model.
HDM are much more complicated allergens than OVA/alum, and our results highlight the need to better
understand the dynamics of SCD lung disease and co-morbid allergic asthma.
120.
Poster 37.
Stress Mediated Immunomodulation: Role of Metallothionein in the extracellular environment
Sadikshya Bhandari, Michael A. Lynes
University of Connecticut, Storrs, CT
Metallothioneins (MTs) are highly conserved stress response proteins, which are up-regulated after
exposure to heavy metals or reactive oxygen and nitrogen species. Of the 61 amino acids present in a con-
sensus mammalian MT sequence, 20 are cysteine residues that are integral to this protein’s biological prop-
erties. Traditionally, research on MT focused on its intracellular roles as a free radical scavenger, metal
reservoir and regulator of NF-kB and Sp-1 transcription factor activities. Though MT lacks a signal peptide
sequence, its presence in extracellular spaces such as milk, bronchoalveolar and prostatic fluids, bile, liver
sinusoids, serum etc. has been well documented. MT has also been detected at sites of inflammation and
tissue wounding. We have previously shown that MT can bind to the surface of lymphocytes, is a potent
chemoattractant for T cells and this effect can be blocked with antagonists of G-protein coupled receptors.
When T cells are pre-incubated with MT, there is a significant and substantial decrease in their chemotac-
tic response towards SDF-1a. This pre-incubation does not interfere with internalization of the cognate
SDF-1a receptor (CXCR4) that accompanies SDF-1a induced chemotaxis. Pre-incubation of MT with T
cells also does not change intracellular reactive oxygen species (ROS) levels. MT can also block SDF-1a
induced chemotactic responses of a both primary splenocytes and a breast cancer cell line, MDA-MB 231
(ATCC HTB-26), suggesting that this is not a cell-specific phenomenon. Our most recent studies have
used specific MT peptides to identify those MT sequences that can interfere with SDF-1a mediated chem-
otaxis. The presence of MT in the extracellular environment and its novel chemotactic properties suggest a
possible immunomodulatory role in inflammatory diseases and in the progression of metastatic cell move-
ment. Hence, MT manipulation under these conditions may provide a possible avenue of therapeutic inter-
vention.
121.
Poster 38.
Commensal microbes drive the generation of systemic IgA responses
Joel R. Wilmore, Brian Gaudette, Wenzhao Meng, Eline T. Luning Prak, and David Allman
Department of Pathology and Laboratory Medicine, Perelman School of Medicine at the University of
Pennsylvania, Philadelphia, PA
It is well known that IgA functions as a critical component of the mucosal barrier in the gut by vir-
tue of its ability to be secreted into the intestinal lumen. However, little is known about the origin, function
and regulation of IgA in the serum. Mucosal IgA responses have been generally thought of as short-lived
and restricted locally to mucosal tissues. However, we find that IgA-secreting PCs make up the majority of
the BM PC pool in standard C57BL/6 mice bred in our colony. The IgA+ BM PCs are predominantly in the
long-lived pool and express gut homing factors such as CCR9 and the integrin α4β7 (alpha4 beta7), sug-
gesting a mucosal origin. The extent to which the commensal microflora influences the BM plasma cell
pool is evident by mice bred in germ free isolators that lack IgA+ PCs in their BM. Additionally, mice from
standard vendors such as Jackson labs (Jax), have extremely low levels of IgA+ PCs in the BM and signifi-
cantly lower serum IgA. Exposing Jax mice to a disparate microflora or Helicobacter sp. led to the genera-
tion of IgA-secreting BM cells, while also inducing increases in serum IgA antibodies enriched for binding
to several commensal bacterial taxa. Moreover, BM IgA-secreting plasma cells exhibited a common clonal
ancestry with intestinal IgA+ plasma cells, and both populations possessed unique gene expression signa-
tures compared to other long-lived BM plasma cells. We conclude that commensal microbes overtly influ-
ence the BM plasma cell pool, and suggest that select commensal microbes can facilitate the induction of
systemic humoral immunity.
122.
Poster 39.
Elucidating the Mechanism of Protective Antibodies Against the Enteric Pathogen Salmonella
Typhimurium at Epithelial Surfaces
Angelene F. Richards1, 2, John J. Varrone1, Jennifer E. Westfall1, and Nicholas J. Mantis1, 2
1Division of Infectious Diseases, Wadsworth Center, New York State Department of Health, Albany, NY
12208; 2Department of Biomedical Sciences, University at Albany School of Public Health,
Albany, NY 12208
Salmonella enterica serovar Typhimurium (ST) is a leading cause of gastroenteritis in the United
States and an emerging cause of invasive non-typhoidal salmonella (NTS) in sub-Saharan Africa. Inva-
sion of the intestinal mucosa by ST involves a complex series of events in which the bacteria utilize flagel-
la-based motility and type 3 secretion systems (T3SS) to adhere to and invade Peyer’s patch tissues before
spreading systemically to the liver and spleen. Here we confirm in an adult mouse model that Sal4, an IgA
monoclonal antibody (mAb) against ST lipopolysaccharide (LPS), blocks bacterial entry into Peyer’s
patch tissues. We show for the first time, however, that an IgG1-derivative of Sal4 when co-gavaged with
ST does not prevent bacterial invasion of Peyer’s patch tissues, even though the same mAb did limit ST
spread to the liver and spleen when administered systemically prior to ST injection into the peritoneal cav-
ity. These results illustrate the importance of IgA in mucosal protection against ST infection and provide a
tractable model to begin to define mechanisms by which secretory antibodies limit bacterial uptake into
Peyer’s patch tissues. With this long goal in mind we generated a second IgA mAb against ST LPS,
named PeA3, which binds an epitope similar to but distinct from Sal4. PeA3 was as effective as Sal4 IgA
at limiting ST infection of mouse Peyer’s patch tissues, indicating that Sal4’s biological activities are not
unique. We have now embarked on a comparative in vitro analysis of Sal4 IgA, Sal4 IgG and PeA3 IgA
mAbs in an effort to define specific properties associated with protective immunity. Sal4 IgA and IgG, as
well as PeA3 IgA, are each potent inhibitors of ST-flagellum based motility in semi-solid agar and liquid
media indicating that they are effective at limiting early steps in the infectious process. However, in vitro
analysis did demonstrate that Sal4 IgA and PeA3 IgA mAbs were more effective than Sal4 IgG at promot-
ing ST agglutination. It has been hypothesized that IgA-mediated agglutination of bacterial pathogens in
the intestinal lumen results in the formation of immune complexes that are readily trapped in mucus and
cleared via peristalsis. Our future goals are to test this hypothesis using in vitro and ex vivo assays, in-
cluding organoids to mimic the intestinal lumen. A better understanding of the mechanisms by which se-
cretory IgA functions in mucosal immunity has important implications for oral vaccine development
against an array of enteric pathogens.
123.
Poster 40.
Macrophages Negatively Regulate Hematopoietic Stem Cells in Murine Aplastic Anemia
Angelica Costello, Amanda McCabe, PhD, Julianne N.P. Smith, PhD, & Katherine C. MacNamara, PhD
Albany Medical College, Albany, NY
Aplastic anemia (AA) is a rare bone marrow (BM) failure syndrome characterized by T cell-
mediated bone marrow destruction and pancytopenia. AA can be genetic or acquired, with the latter caused
by such insults as radiation, chemicals, drugs, and infection. T cell-derived interferon-γ (IFN-γ) has been
implicated in driving disease, but the mechanisms of IFN-γ-mediated pathogenesis in hematopoietic failure
during AA are not well understood. We recently identified a role for IFN-γ signaling specifically in macro-
phages (Mϕs) in driving the transient loss of hematopoietic stem cells (HSCs) in murine ehrlichiosis. Here,
in a mouse model of AA, we find that BM-resident Mϕs are maintained in the BM despite the loss of other
hematopoietic cell types, and that this requires IFN-γ. Moreover, Mϕ depletion with clodronate liposomes
or abrogation of IFN-γ signaling in Mϕs during AA rescues the HSC pool and significantly improves sur-
vival. We observed similar numbers of T-bet+ T cells and comparable IFN-γ production in AA mice that
were depleted of Mϕs, suggesting that Mϕs are critical sensors of IFN-γ and drivers of disease during AA.
Despite this, we did not detect a significant reduction in inflammatory proteins when IFN-γ signaling is ab-
rogated or when Mϕs were depleted. The one notable exception, however, was the chemokine CCL5
(RANTES), which was highly expressed in the BM during AA and significantly reduced in mice depleted
of Mϕs or when Mϕs were unable to respond to IFN-γ. CCL5 production in BM fibroblastic cells is in-
duced via podoplanin (PDPN) signaling. In support of a role of PDPN signaling in driving increased CCL5
during AA, we detected a significant increase in PDPN+ BM-resident Mϕs during AA. Neutralization of
either CCL5 or PDPN during AA rescued HSC numbers. Moreover, anti-PDPN treatment rescued platelet
numbers and significantly improved survival. Altogether, we demonstrate a novel role for IFN-g in AA
pathogenesis whereby IFN-γ promotes increased PDPN+ Mϕs and enhanced production of CCL5, thus
driving HSC loss and thrombocytopenia.
124.
Poster 41.
Neutralizing Camelid Antibodies Target the Binding Subunit of Ricin by Interfering with
Receptor Attachment and Intracellular Trafficking
Amanda Y. Poon, David J. Vance, Yinghui Rong, Nicholas J. Mantis Wadsworth
Center, New York State Department of Health, Albany NY 12208 Department of Bio-
medical Sciences, University at Albany, Albany NY 12208
Ricin toxin is a ribosome-inactivating protein (RIP) and classified by the Center for Disease Control
and Prevention as a potential bioterrorism agent. Ricin’s B-subunit (RTB) is a lectin that mediates toxin
attachment, internalization, and intracellular trafficking of ricin in mammalian cells, and is therefore
indispensable for ricin’s cytotoxic activity. RTB (32 kDa) consists of two homologous domains (1 and
2), each divided into three subdomains (a, b, g), with lectin activity restricted to subdomains 1a
and 2g.
Fig.1: Linear depiction of RTB
Theoretically, blocking RTB binding and/or trafficking activity is an ideal means by which to neu-
tralize ricin. However, analysis of dozens of RTB-specific monoclonal antibodies (mAbs) indicate
that toxin-neutralizing antibodies are extremely rare and likely target a very limited number of
epitopes, possibly in subdomains 1a and 2g. To better understand RTB-antibody interactions we
have embarked on a concerted effort to generate a comprehensive B cell epitope map of RTB using a
highly diverse ricin-specific camelid single domain antibody (VHH) library. Currently, we have isolated
a panel of RTB-specific VHHs with a range of affinities and in vitro toxin-neutralizing activities. Using
a novel biotinylated-ricin competition ELISA, we observed that VHHs with the best toxin-neutralizing
activity tend to recognize epitopes within the vicinity of subdomain 2g, whereas VHHs with weak neu-
tralizing activity cluster near 1a. We next tested the hypothesis, using a solid phase receptor bind-
ing assay, as well as THP-1 cells in a flow cytometric based readout, that ricin neutralizing activity
correlates with the ability of VHHs to interfere with toxin attachment to mammalian cell surfaces. The
hypothesis proved incorrect, as neutralizing and non-neutralizing VHHs interfered with ricin attach-
ment to similar degrees. Based on these and other results, we are now investigating the possibility that
toxin-neutralizing activity by RTB-specific VHHs is not due to interference of cell attachment, but rather
effects on toxin endocytosis and/or intracellular trafficking. At the same time we have initiated efforts
to perform more detailed epitope mapping studies by X-ray crystallography (X-TAL) and hydrogen-
deuterium exchange coupled with mass spectrometry (HX-MS). As RTB is the prototypic member
of a large family of toxin and bacterial R-type lectins we expect our results will afford insights into
basic mechanisms of pathogen-host interactions.
125.
Poster 42.
The lymphoproliferative effects of the small stress response protein, metallothionein
Kristen E. Dostie, Michael A. Lynes.
University of Connecticut, Storrs CT
Metallothioneins are small metal-binding proteins that can alter a variety of immune responses,
and, for example, can influence the progression of autoimmune diseases and the response to bacterial in-
fection. MTs are biochemically unique proteins due to their low molecular weight and unusually high cys-
teine content, which accounts for ~33 mol% of the total amino acid composition. These cysteines within
MT provide sulfhydryl groups which bind essential metal cations such as zinc and copper and can also
serve to regulate the local redox environment. Although MT lacks an N-terminal signal peptide and has
conventionally been considered an intracellular protein, it has been found in extracellular compartments
where it can act as a potential “danger signal” that can mediate both innate and adaptive immune respons-
es. Previous work of our laboratory has shown that MT binds to the surface of lymphocytes and can lead to
proliferation of lymphocytes. Interestingly, MT can synergistically enhance the proliferative response of
lymphocytes when co-administered with either T- or B-cell mitogens. The aim of this research is to further
characterize the mechanism responsible for the enhanced proliferative response to MT of various lympho-
cyte subsets. Our work profiles the relative binding affinity of MT to surface molecules of specific lym-
phocyte populations. These relative binding affinities of MT for different lymphocyte populations may
provide insight to potential MT receptors on the cell surface.
126.
Poster 43.
The Role of Pseudomonas aeruginosa Metallothionein, PmtA, in Immune Modulation
Amy Thees, Kathryn Pietrosimone, Brian Greco, Michael A. Lynes
Pseudomonas aeruginosa is a gram-negative, opportunistic pathogen, frequently associated with hospital-
acquired infections. Chronic infections of Pseudomonas aeruginosa are common in cystic fibrosis patients
due to exaggerated and ineffective airway inflammatory processes that fail to eradicate this pathogen. A
novel bacterial stress protein, PmtA, has been shown to contribute to bacterial survival by influencing the
host immune response. Disruption of the PmtA gene production causes a decrease in the production of a
redox-active, secondary metabolite, pyocyanin. It has also been demonstrated that PmtA blocks the migra-
tion of cells in response to SDF-1 alpha and decreases SDF-1 alpha-induced internalization of CXCR4 on
Jurkat T cells. Here we show that PmtA can be found on the surface of leukocytes in culture after expo-
sure to exogenous MT, which suggests that leukocytes may express one or more receptors for PmtA. Ex-
posure to PmtA also enhances proliferation of Jurkat T cells. Pseudomonas aeruginosa is known for the
multitude of virulence factors that it can produce. PmtA may represent an additional tool that Pseudomo-
nas aeruginosa can employ to sustain an infection.
127.
Poster 44.
Regulation of IgM Memory B cell Pool Size by the Inhibitory receptor FcgRIIb
Amber Papillion and Gary Winslow
Upstate Medical University, Syracuse, NY
Ehrlichia muris infection generates a long-term CD11c/T-bet-positive IgM memory population in
the spleen (J. Immunol. 191:1240). Among the many surface markers that distinguish the IgM memory
cells from canonical B cells is the inhibitory Fc receptor, FcgRIIb, which exhibited a 2-fold higher expres-
sion. We hypothesized FcgRIIb negatively regulates IgM memory cells by binding immune complexes
present during low-level chronic infection. To investigate this question, we monitored the IgM memory
cell population in infected FcgRIIb-deficient mice. Thirty days post-infection, the IgM memory B cells
were generated earlier, and were found at three-fold higher frequencies in FcgRIIb-deficient mice, com-
pared to wild-type mice. This increase in the frequency of spleen IgM memory cells was due to an increase
in cell number, and was in turn associated with an increase in antigen-specific IgG. These data indicate
that FcgRIIb plays an important role in regulating the expansion and/or persistence of IgM memory cells in
wild-type mice under conditions where antigen-specific IgG is sufficient to control infection. Other studies
revealed that the IgM memory population in FcgRIIb-deficient mice exhibited much lower expression of
FAS, CD40, BAFF-R, and TACI. We therefore proposed that FcgRIIb signaling, likely via immune com-
plexes, acts in wild-type cells to regulate the size of the IgM memory cell pool, by maintaining the expres-
sion of FAS and other receptors that regulate cell survival. These data suggest a novel regulatory role for
FcgRIIb in B cell memory
128.
Poster 45.
Skin Microbiome and the Development of Autoimmune Vitiligo
Jennifer Vella, Walburga Croteau, Yina Huang
Dartmouth College, Hanover NH
Skin cancer is the most common form of cancer and can be classified into two groups: melanoma
and non-melanoma, with melanoma being the more dangerous of the two. Melanoma arises when our pig-
ment producing cells, called melanocytes, turn cancerous, gaining the ability to rapidly divide and metasta-
size to other organs throughout the body. Melanoma patients who simultaneously develop autoimmune
vitiligo have a more favorable prognosis, suggesting therapies increasing vitiligo may be beneficial. While
studies have shown that autoimmune vitiligo is a CD8 T cell mediated disease, the mechanisms responsi-
ble for its development remain unclear. By utilizing a mouse model we are able to mimic melanoma-
induced vitiligo allowing us to study potential triggers of this autoimmune disease. With this method we
compared the development of vitiligo in response to various treatments. We found that mice treated with
broad-spectrum antibiotics were less likely to develop vitiligo compared to their untreated littermates. In
addition to the decrease vitiligo incidence, the mice that did develop vitiligo only developed it locally,
without dissemination of depigmentation. Our studies indicate that the skin microbiome may play an im-
portant role in the development and progression of autoimmune vitiligo.
129.
Poster 46
Hypofractionated vs. hyperfractionated radiotherapy treatment alters immune cell viability and
function
Margaret L. Barlow1, Nicholas Battaglia1, Scott A. Gerber1, and Edith M. Lord1
1Department of Microbiology and Immunology, University of Rochester, Rochester, NY 14642
Radiotherapy is one of the most effective means of treating solid tumors in cancer patients. This is
due to the radiation acting in two ways to control tumor growth. First, by inducing cell death due to direct
or indirect damage to the DNA, then by the killed tumor cells releasing tumor antigens and inflammatory
signals that boost the antitumor immune response. Unfortunately, radiotherapy may also damage the
healthy tissue surrounding the tumor, causing adverse side effects in patients. To limit this damage, radio-
therapy in the clinic is generally delivered as multiple small doses (hyperfractionation) over the course of
several weeks. However, in preclinical radiotherapy studies, radiation is often given as a single or a few
high ablative doses (hypofractionation). We previously observed improved tumor growth control and an
increased number of tumor infiltrating lymphocytes (TIL) in tumors given a single ablative dose when
compared to a hyperfractionated scheme. As TILs are radiosensitive, we hypothesize that the repeated dos-
es of a hyperfractionated scheme may result in increased killing of the TILs as they infiltrate the tumor,
damaging them as they attempt to carry out their effector response and thereby reducing the effectiveness
of the anti-tumor immune response. To further investigate this, we created two radiation dosing schedules
based on current clinical protocols. Our hypofractionated scheme used either one or two doses of 10 Gy,
while our hyperfractionated scheme used five doses of 2 Gy. We compared the responses of the mouse
Colon38 tumor model to these schemes. In vitro, the hypofractionation treatment was more effective at
killing tumor cells and inducing the tumor cells to produce the inflammatory cytokines IL-β(beta), TNFa
(alpha), IL-6, as well as the chemoattractant IP-10. In vivo, the hypofractionated treatment resulted in a sig-
nificant delay in tumor outgrowth and improved survival rates when compared to the hyperfractionated
treatment. We also observed fewer apoptotic or dead effector CD8+ T cells before the second dose of 10 Gy
in the hypofractionated scheme, compared to 24 hours after the second 10 Gy dose, suggesting that some
of these cells were damaged by the repeated radiation. Therefore, hypofractionated radiotherapy may in-
crease immunity and preserve immune cell viability and function. Funded by NIH R01CA28332 and
T32AI007285.
130.
Poster 47.
The role of Indoleamine 2,3-dioxygenase (IDO) in the survival of bone marrow resident long lived
Plasma cells
Shivana M. Maharaj, Louise M. Carlson, and Kelvin P. Lee
Department of Immunology, Roswell Park Cancer Institute; Buffalo, NY
Long lived plasma cells (LLPCs) are essential for sustained antibody responses and protective hu-
moral immunity. How these cells maintain longevity and a durable antibody response is largely dependent
on the complex nature of the bone marrow microenvironment in which these cells reside, and the pro sur-
vival factors produced in this niche. Work in our lab has shown that CD28 is required for LLPC survival,
and that Indoleamine 2,3-dioxygenase (IDO), an enzyme that catabolizes tryptophan, is produced as a re-
sult of back signaling from CD28:CD80/86 interactions between plasma cells and bone marrow myeloid
derived DCs. IDO is classically known to contribute to an immunosuppressive environment through its
depletion of tryptophan (specifically with respect to T cells). However, to our great surprise we found that
there are fewer LLPCs present in the bone marrow of IDO knockout mice in comparison to wild type
mice. Additionally, live cell numbers of purified plasma cells in vitro in tryptophan free media and IDO
conditioned media are greater than in media alone. These conditions also caused autophagy to occur in
these plasma cells, a process shown to be required for LLPC survival. This leads us to propose a model
where CD28, through back signaling to CD80/86, induces IDO production which leads to depletion of
tryptophan and causes induction of autophagy, a mechanism LLPCs use to compete and survive within
the bone marrow microenvironment.
Funding: T32 CA085183
131.
Poster 48.
Pretreatment Peripheral Blood Monocyte Subset Signature is Predictive of Patient Response to
Dendritic Cell Vaccination
Anand Sharda1, Alexander Wald1
, Mohammad Habiby Kermany1, Katja Koeppen2
, Thomas Hampton2, Jan
Fisher3, Camilo Fadul3
, Marc Ernstoff4, Thomas Schwaab1
, Jason Muhitch1,5
Department of Urology1, Medicine4
, and Immunology5 Roswell Park Cancer Institute, Buffalo, NY Depart-
ments of Microbiology and Immunology2, and Medicine3 Geisel School of Medicine at Dartmouth,
Hanover, NH
Clinical trials have demonstrated that dendritic cell (DC) vaccination can initiate durable anti-tumor
immunity in a subset of cancer patients resulting in complete responses, even in stage IV Renal Cell Carci-
noma (RCC). The influence of monocytes, the starting material for conventional DC vaccines, on patient
responses remains under-investigated. Recently, three subsets of monocytes have been described (classical,
intermediate, and non-classical), each with distinct functional properties. However, their roles in anti-
tumor immunity, particularly in the context of DC vaccination, are unclear. The goal of this study was to
determine whether the circulating pretreatment monocyte subset gene expression and composition from
Stage IV RCC patients prior to DC vaccination predicted responses to treatment in a completed phase II
clinical trial (NCT00085436). Pretreatment circulating classical (CD14++, CD16-), intermediate (CD14++,
CD16+), and non-classical (CD14+, CD16++) monocyte subsets were isolated from patients. Pretreatment
peripheral blood from complete responders (2 of 3 have no observable disease > 5 years following therapy)
contained fewer classical monocytes (57.5% ± 6.4) compared to all other groups (P < 0.05). Interestingly, a
higher percentage of DC derived from non-classical monocytes expressed costimulatory molecules (CD80;
96.4%, CD86; 91.1%, HLA-DR; 99.1%) compared to classical monocyte-derived DC (64.6%, 54.4%,
60.2%, respectively). DC derived from non-classical monocytes were also superior in their ability to in-
duce allogeneic T cell proliferation compared to DC originating from classical monocytes. Additional gene
expression analysis by unsupervised hierarchical clustering clearly distinguished the transcriptional profile
of classical, intermediate, and non-classical monocytes from RCC patients to healthy controls. Further in-
vestigation revealed that monocytes from long term survivors (> 10 years) could be distinctly segregated
from other RCC patients. These findings demonstrate that DC-derived from the minor CD16+ monocyte
subset may represent a superior product for use in vaccination protocols. Gene expression profiling of cir-
culating monocytes may provide an accessible biomarker for patient responsiveness to immunotherapy.
Future studies will address whether pretreatment levels of intermediate and non-classical monocytes are
prognostic indicators for response to additional immunotherapies, including checkpoint blockade inhibi-
tors.
132.
Poster 49.
Antagonistic Control of Intestinal Wnt Expression by IBD-Associated Cytokines
Travis Walrath, Stephen Sharp M.D., Shanti D'Souza, Prabhu Tewari, and William O'Connor Jr, Ph. D.
Albany Medical College, Albany NY
Homeostasis of the intestinal epithelium depends on the complex interplay of a variety of mediators
including cytokines that control, among other processes, a gradient of Wnt proteins. Homeostasis can be
disrupted by acute and chronic inflammation such as Inflammatory Bowel Disease (IBD). During IBD a
high concentration of the TH1 and TH17 associated cytokines, interferon gamma (IFN-γ) and interleukin-
17A (IL-17A) respectively, is observed within the colonic lamina propria. IFN-γ has previously been
shown to be cytotoxic to the intestinal epithelium, in part through interference with canonical Wnt signal-
ing; this is associated with increased disease severity in IBD. In murine models of disease, IL-17A has
been shown to lessen the severity of induced IBD.
One important question in the field is how these cytokines govern the intestinal epithelial and adja-
cent lymphoid compartments to either amplify inflammation or support mucosal healing. Toward under-
standing how these cytokines regulate the colonic microenvironment, we undertook a study using murine
knockout strains, primary tissue treated ex vivo, and primary spheroid/organoid cultures treated with IFN-γ,
IL-17A, or a combination of both. We found that IFN-γ reduced canonical Wnt expression while inducing
expression of non-canonical Wnts in primary murine colon tissue. Additionally we found that this altered
balance of wnt expression is dependent on the presence of lymphocytes and further, that wnt5a is specifi-
cally induced in CD19+ cells in the colon. Interestingly, IL-17A was able to oppose the increased expres-
sion of wnt5a. Moreover, mice deficient in IL-17A exhibit enhanced colonic wnt5a and TH1 cell marker
expression in response to Citrobacter rodentium infection, suggesting that IL-17A is opposing infection-
induced wnt5a and the TH1 bias during active colonic inflammation.
Canonical wnts, such as Wnt3a, are necessary for maintaining the potency of stem cells in the in-
testinal stem cell niche and the proliferative capacity of the intestinal epithelium. Non-canonical wnts such
as Wnt5a, have been shown to inhibit canonical wnt signaling and support tissue damage during colitis by
supporting increased IFN-γ production. Therefore, our data suggest that IFN-γ promotes intestinal epitheli-
al dysfunction in part by promoting a shift from canonical to non-canonical Wnt expression and function in
the colon. Furthermore, we hypothesize that IL-17A interferes with the IFN-g mediated shift in Wnt ex-
pression, and therefore may reduce IFN-g mediated epithelial toxicity and facilitate mucosal healing.
133.
Authors Index
CS – Corporate Speaker P - Poster Number
KS - Keynote Speaker S – Speaker
O - Oral Poster Presentation WP – Workshop Presenter
Page (s)
*************************************************************************************
Abrams, Scott I. ................................................................................................................... 50, 52, 97, 107, 110
Allman, David ..................................................................................................................... 59, 119
Amiel, Eyal .......................................................................................................................... 64, 104
Amobi, Adaobi (P26, O) ...................................................................................................... 51, 109
Appenheimer, Michelle ....................................................................................................... 52, 110
Arend, Kyle ......................................................................................................................... 85
August, Avery ...................................................................................................................... 44, 89
Baranova, Danielle E. (P34) ................................................................................................ 117
Barbachano-Guerrero, Arturo (P31) .................................................................................... 79, 114
Barbi, Joseph (S) .................................................................................................................. 72
Barlow, Margaret L. (P46) ................................................................................................... 129
Battaglia, Nicholas ............................................................................................................... 129
Benoit, Patrick ..................................................................................................................... 47, 103
Besra, Gurdyal ..................................................................................................................... 46, 100
Bhandari, Sadikshya (P37) .................................................................................................. 120
Bhat, Tariq A. (P16) ............................................................................................................ 99
Blader, Ira J. (S) ................................................................................................................... 81
Bogner, Paul N. ................................................................................................................... 99
Boule, Lisbeth A. ................................................................................................................. 111
Boyson, Jonathan E. ............................................................................................................ 47, 103
Braciale, Thomas J. (WP, KS) ............................................................................................. 76, 82
Budd, Ralph ......................................................................................................................... 73
Burke, Catherine G.(P28) .................................................................................................... 111
Buskiewicz, Iwona A. (S) .................................................................................................... 73
Carlson, Louise M. .............................................................................................................. 48, 58, 106, 130
Carlson, Bradley A. ............................................................................................................. 102
Chan, Gary C. ...................................................................................................................... 85, 108
Chapman, Timothy J. (S) ..................................................................................................... 74, 116
Consiglio, Camila Rosat (P18, O) ....................................................................................... 63, 101
Cojohari, Olesea (P25) ........................................................................................................ 85, 108
Cooper, James ...................................................................................................................... 58, 106
Costello, Angelica (P40, O) ................................................................................................. 65, 123
Coupet, Tiffany (P32) .......................................................................................................... 115
Croteau, Walburga ............................................................................................................... 128
134.
Cubitt, Rebecca L. ................................................................................................................ 43, 86, 88
D’Alessandro, Angelo .......................................................................................................... 64, 104
Davenport, Miles P. ............................................................................................................. 40
DeVault, Victoria L. (P20, O) .............................................................................................. 47, 103
Diehl, Michael ..................................................................................................................... 50, 107
Dienz, Oliver ........................................................................................................................ 47, 103
Dostie, Kristen E. (P42) ....................................................................................................... 125
D’Souza, Shanti ................................................................................................................... 66
Elliott, Michael R. ................................................................................................................ 93
Emo, Jason A. (P7) .............................................................................................................. 74, 90
Endy, Timothy ..................................................................................................................... 114
Enelow, Richard I. (S) ......................................................................................................... 79
Ernstoff, Marc ...................................................................................................................... 131
Evans, Sharon S. .................................................................................................................. 50, 52, 107, 110
Fadul, Camilo ....................................................................................................................... 53, 131
Fino, Kristin ......................................................................................................................... 98
Fisher Jan ............................................................................................................................. 53, 131
Gaudette, Brian .................................................................................................................... 59, 121
Gerber, Scott A. ................................................................................................................... 112, 129
Georas, Steve N. .................................................................................................................. 74, 116
Ginty, Fiona (S) ................................................................................................................... 71
Gollnick, Sandra .................................................................................................................. 63, 101
Greco, Brian ......................................................................................................................... 126
Green, Kathy A. (P30) ......................................................................................................... 113
Green, William R. ............................................................................................................... 113
Grimson, Andrew ................................................................................................................. 40
Hägglöf, Thomas .................................................................................................................. 46, 100
Hampton, Thomas ................................................................................................................ 53, 131
Harton, Jonathan A. ............................................................................................................. 92
Haque, Mohammad ............................................................................................................. 98
Hartley, Richard .................................................................................................................. 73
Henry, Everett ...................................................................................................................... 86
Hillman, Sara E. ................................................................................................................... 74, 116
Huang, Weishan (P6, O) ....................................................................................................... 44, 89
Huang, Yina ......................................................................................................................... 128
Hyuck, Heidie ...................................................................................................................... 90
Jiang, Chenyang (P36) ......................................................................................................... 119
Kalathil, Suresh G. ............................................................................................................... 99
Karlsson, Mikael .................................................................................................................. 46, 100
Katkere, Bhuvana (P19) ....................................................................................................... 102
135.
Kearsing, Lori ...................................................................................................................... 41
Kelly, Ryan .......................................................................................................................... 73
Kenderes, Kevin (P22, O) .................................................................................................... 57, 105
Kermany, Mohammad Habiby ............................................................................................ 53, 131
Khan, ANM Nazmul H. ....................................................................................................... 49, 87
King, Christine A. (S) .......................................................................................................... 79, 114
Kiramanjeswara, Girish S. ................................................................................................... 102
Knowlden, Sara A. ............................................................................................................... 73
Koch, Cameron J. ................................................................................................................ 111
Koenig, Andreas .................................................................................................................. 73
Koeppen, Katja .................................................................................................................... 53, 131
Koylass, Nicholas ................................................................................................................ 44, 89
Ku, Amy W. (P27, O) .......................................................................................................... 50, 52, 107, 110
Lang, Sabine ........................................................................................................................ 84
Lanthier, Paula ..................................................................................................................... 46, 100
Lawrence, B. Paige .............................................................................................................. 45, 94, 111
Lawrence, David A. (S) ....................................................................................................... 41
Leadbetter, Elizabeth .......................................................................................................... 46, 100
Lee, Kelvin P. ...................................................................................................................... 48, 58, 106, 130
Levinson, Nicholas J. ........................................................................................................... 117
Lilley, Graham W.J. ............................................................................................................ 47, 103
Lord, Edith M. ..................................................................................................................... 112, 129
Lubliner, Jane ...................................................................................................................... 41
Lugade, Amit A. .................................................................................................................. 99
Lynes, Michael A. (S) .......................................................................................................... 36, 120, 125, 126
MacNamara, Katherine C. ................................................................................................... 65, 123
Maharaj, Shivana M. (P47) .................................................................................................. 130
Mahpour, Amin ................................................................................................................... 58, 106
Mantis, Nicholas J. .............................................................................................................. 117, 122, 124
Markley, Rachael L. ............................................................................................................ 102
Martinez, Natalie ................................................................................................................. 90
Massa, Paul T. ..................................................................................................................... 62, 91
McCabe, Amanda ................................................................................................................ 65, 123
Mendoza, Alvaro ................................................................................................................. 41
Meng, Wenzhao ................................................................................................................... 59, 121
Metzger, Dennis W. ............................................................................................................. 95, 118
Miller, A. ............................................................................................................................. 97
Minchenberg, Scott B. (P8, O) ............................................................................................ 62, 91
Minderman, Hans ................................................................................................................ 50, 107
Mohrs, Katja ........................................................................................................................ 46, 100
136.
Mondal, Tapan .................................................................................................................... 41
Montgomery, Theresa ......................................................................................................... 73
Moon, Taylor J. (P10) ......................................................................................................... 93
Moorman, Nathaniel ............................................................................................................ 85
Moshkani, Safiehkhatoon (P1) ............................................................................................ 84
Moysich, Kirsten B. ............................................................................................................ 49, 87
Muhitch, Jason ................................................................................................................... 50, 52, 53, 107, 110, 131
Murphy, Michael ................................................................................................................ 73
Murthy, Aditi (P29) ............................................................................................................. 112
Myers, Jason R. ................................................................................................................... 45, 94, 111
Nagar, Abhinit (P9) ............................................................................................................. 92
Nemeth, M. .......................................................................................................................... 97
Noelle, Randolph J. ............................................................................................................. 113
O’Connor, William .............................................................................................................. 66, 132
Odunsi, Kunle ...................................................................................................................... 50, 51, 87, 109
O’Loughlin, Kieran ............................................................................................................. 50, 107
Ostrand-Rosenberg, Suzanne .............................................................................................. 50, 107
Oyesola, Oyebola (P5, O) .................................................................................................... 43, 88
Papillion, Amber (P44, O) ................................................................................................... 57, 60, 105, 127
Palmer, Claire (P32) ............................................................................................................ 115
Pan, Fan .............................................................................................................................. 70
Pardoll, Drew ...................................................................................................................... 70
Paredes, Anthony F. ............................................................................................................ 62, 91
Peng, Peng ........................................................................................................................... 58, 106
Peppenelli, Megan A. (P2) .................................................................................................. 85, 108
Perl, Andras ......................................................................................................................... 73
Phipps, Richard P. ............................................................................................................... 99
Pietrosimone, Kathryn ......................................................................................................... 126
Poon, Amanda Y. (P41) ...................................................................................................... 124
Post, Christina M. (P11, O) ................................................................................................. 45, 94
Powers, Colin A. (P24, O) ................................................................................................... 50, 107
Prabhu, Kumble S. .............................................................................................................. 102
Prak, Eline T. Luning .......................................................................................................... 59, 121
Pryhuber, Gloria .................................................................................................................. 90
Rahman, Tabassum ............................................................................................................. 92
Rahman, Ziaur S.M. ............................................................................................................ 56, 96
Reynaldi, Arnold ................................................................................................................. 40
Richards, Angelene F. (P39) ............................................................................................... 122
Robek, Michael D. .............................................................................................................. 84
Rodriguez, Princess ............................................................................................................. 64, 104
137.
Rong, Yinghui ..................................................................................................................... 124
Rose, John K. ....................................................................................................................... 84
Rudd, Brian D. (S) ............................................................................................................... 40
Sandhu, Praneet Kaur (P15) ................................................................................................ 98
Scheible, Kristin .................................................................................................................. 90
Schwaab, Thomas ............................................................................................................... 53, 131
Schell, Stephanie L. (P13, O) .............................................................................................. 56, 96
Schneider, Karin (S) ............................................................................................................ 38
Schwartzberg, Pamela L. ..................................................................................................... 47, 103
Segal, Brahm H. ................................................................................................................... 49, 87
Sentman, Charles L. ............................................................................................................. 115
Sharda, Anand P. (P48, O) ................................................................................................... 50, 53, 107, 131
Sharp, Stephen ..................................................................................................................... 66, 132
Shay, Ashley E. ................................................................................................................... 102
Sime, Patricia J. ................................................................................................................... 99
Singel, Kelly L. (P4, O) ....................................................................................................... 49, 87
Siracusa, Mark C. ................................................................................................................ 86
Skitzki, Joseph J. ................................................................................................................. 50, 107
Smith, Julianne N.P. ............................................................................................................ 65, 123
Smith, Norah L. ................................................................................................................... 40
Smyth, Tim .......................................................................................................................... 116
Solouki, Sabrina ................................................................................................................... 44, 89
Song, Jianxun ..................................................................................................................... 98
Soni, Chetna ........................................................................................................................ 56, 96
Soucy, Alicia (P12) .............................................................................................................. 95
Steiner, Donald (P35) .......................................................................................................... 118
Szczepanek, Steven M. ........................................................................................................ 119
Tewari, Prabhu .................................................................................................................... 66, 132
Thanavala, Yasmin .............................................................................................................. 99
Thatcher, Thomas H. ........................................................................................................... 99
Thees, Amy (P43) ................................................................................................................ 126
Thwe, Phyu (P21, O) ........................................................................................................... 64, 104
Topham, David .................................................................................................................... 90
Twum, Danielle Y.F. (P14) ................................................................................................. 97
Utley, Adam (P23, O) .......................................................................................................... 48, 58, 106
Vance, David J. .................................................................................................................... 124
Varrone, John J. ................................................................................................................... 122
Veazey, Janelle (P33) .......................................................................................................... 116
Veerapen, Natacha ............................................................................................................... 46, 100
Vella, Jennifer (P45) ............................................................................................................ 128
138.
Ventro, Daniela .................................................................................................................... 48
Vignali, Paolo D.A. ............................................................................................................. 70
Vomhof-Dekrey, Emilie ...................................................................................................... 46, 100
Wald, Alexander ................................................................................................................. 53, 131
Walrath, Travis (P49, O) ..................................................................................................... 66, 132
Wang, Jocelyn ..................................................................................................................... 40
Wang, Li .............................................................................................................................. 113
Watson, Neva ...................................................................................................................... 40
Webb, Lauren M. (P3) ......................................................................................................... 43, 86, 88
Westfall, Jennifer E. ............................................................................................................ 122
Williamson, David R. .......................................................................................................... 102
Wilmore, Joel R. (P38, O) ................................................................................................... 59, 121
Winslow, Gary ..................................................................................................................... 57, 60, 105, 127
Wissink, Erin ....................................................................................................................... 40
Wojno, Elia Tait ................................................................................................................... 43, 86, 88
Yang, Qi (S) ........................................................................................................................ 37
Yates, Jennifer (P17, O) ...................................................................................................... 46, 100
Yokoyama, Wayne M. (KS, WP) ........................................................................................ 34, 67
Yu, Hong ............................................................................................................................. 70
Zhang, Kangning ................................................................................................................. 37
139.
Amiel, Eyal
University of Vermont
802-656-0522
Amobi, Adaobi
Roswell Park Cancer Institute
716-583-0875
Baranova, Danielle
Wadsworth Center
518-473-3947
Barbachano-Guerrero, Arturo
SUNY Upstate Medical University
315-464-9490
Barlow, Margaret
University of Rochester
216-410-5140
Battaglia, Nicholas
University of Rochester
716-471-6114
Bellville, Dawn
Albany Medical College
518-262-5365
Berwin, Brent
Dartmouth College
603-208-7446
Bhandari, Sadikshya
University of Connecticut
580-647-0228
Bhat, Tariq
Roswell Park Cancer Institute
716-602-9907
Blader, Ira
University at Buffalo
716-852-9580
Braciale, Thomas
University of Virginia
434-825-2415
Burke, Catherine
University of Rochester
716-949-2802
Buskiewicz, Iwona
University of Vermont
802-6568507
Bynoe, Margaret
Cornell University
607-253-4023
Chapman, Timothy
University of Rochester
585-314-3935
Attendee Contact Information
140.
Cojohari, Olesea
SUNY Upstate Medical University
315-944-5434
Consiglio, Camila Rosat
Roswell Park Cancer Institute
716-845-8167
Costello, Angelica
Albany Medical College
518-262-0922
Coupet, Tiffany
Dartmouth College
571-393-1357
DeVault, Victoria
University of Vermont
978-895-3204
Dostie, Kristen
University of Connecticut
860-486-3648
Drake, James (Jim)
Albany Medical College
518-262-9337
Elliott, Michael
University of Rochester
585-273-4793
Emo, Jason
University of Rochester
585-273-1408
Enelow, Richard
Dartmouth College
802-356-9653
Ginsberg, Arielle
BD Biosciences
347-205-5493
Ginty, Fiona
GE Global Research
518-366-6465
Gondre-Lewis, Timothy
NIAID/NIH
240-627-3566
Green, Kathy
Dartmouth College
603-650-5056
Green, William
Dartmouth College
603-650-8607
Harton, Jonathan
Albany Medical College
518-262-4445
141.
Howell, Tyger
Roswell Park Cancer Institute
716-845-8231
Huang, Weishan
Cornell University
607-253-4014
Huang, Yina
Dartmouth Medical College
603-650-7545
Jiang, Chenyang
University of Connecticut
860-771-9028
Katkere, Bhuvana
Pennsylvania State University
518-339-8133
Kenderes, Kevin
SUNY Upstate Medical University
607-341-1511
King, Christine
SUNY Upstate Medical University
315-464-5465
Ku, Amy
Roswell Park Cancer Institute
661-618-0258
Lawrence, David
Wadsworth Center/SUNY Albany
518-486-9154
Lord, Edith
University of Rochester
585-749-8604
Lozito, Shannon
University of Vermont
802-258-0064
Lynes, Michael
University of Connecticut
860-486-4350
MacNamara, Kate
Albany Medical College
518-262-0921
Maharaj, Shivana
Roswell Park Cancer Institute
716-380-0223
Mantis, Nicholas
Wadsworth Center/SUNY Albany
518-473-7487
Metzger, Dennis
Albany Medical College
518-262-6750
142.
Minchenberg, Scott
SUNY Upstate Medical University
516-524-5812
Moon, Tayler
University of Rochester
315-796-7728
Moshkani, Safie
Albany Medical College
518-264-2588
Muhitch, Jason
Roswell Park Cancer Institute
716-845-4930
Murthy, Aditi
University of Rochester
508-813-3363
Nagar, Abhinit
Albany Medical College
518-262-4447
Norris, Carol
University of Connecticut
860-486-3648
O’Connor, William
Albany Medical College
518-262-6548
Oyesola, Oyebola
Cornell Univesity
607-379-7745
Palker, Thomas
NIAID/NIH
301-828-7192
Palmer, Claire
Dartmouth College
802-730-2807
Papillion, Amber
SUNY Upstate Medical University
337-580-1924
Pelletier, Marianne
Astra Zeneca
518-872-1582
Peppenelli, Megan
SUNY Upstate Medical University
315-464-7682
Poon, Amanda
Wadsworth Center
347-868-8796
Post, Christina
University of Rochester
781-974-9431
143.
Powers, Colin
Roswell Park Cancer Institute
443-889-6314
Price, B. Duane
NIAID/NIH
240-669-5074
Ravi, Swetha
Pennsylvania State University
313-788-2486
Richards, Angelene
Wadsworth Center
518-420-8281
Robek, Michael
Albany Medical Center
518-264-2580
Rudd, Brian
Cornell University
607-253-4418
Praneet Kaur Sandhu
Pennsylvania State University
716-418-4355
Schell, Stephanie
Pennsylvania State University
443-243-9751
Schneider, Karin
SUNY Upstate Medical University
315-559-8953
Sharda, Anand
Roswell Park Cancer Institute
716-845-7180
Singel, Kelly
Roswell Park Cancer Institute
716-845-3138
Snyder, James
NIAID/NIH
240-669-5060
Soucy, Alicia
Albany Medical College
518-262-6220
Steiner, Donald
Albany Medical College
518-262-6220
Szczepanek, Steven
University of Connecticut
860-486-8101
Thanavala, Yasmin
Roswell Park Cancer Institute
716-845-8536
144.
Thees, Amy
University of Connecticut
732-232-4739
Thwe, Phyu
University of Vermont
414-241-3367
Twum, Danielle
Roswell Park Cancer Institute
716-845-3352
Utley, Adam
Roswell Park Cancer Institute
336-847-4725
Veazey, Janelle
University of Rochester
585-519-8472
Vella, Jennifer
Dartmouth College
603-650-7546
Walrath, Travis
Albany Medical College
518-262-6341
Webb, Lauren
Cornell University
617-763-7499
Wilmore, Joel
University of Pennsylvania
315-882-2959
Winslow, Gary
SUNY Upstate Medical University
315-464-7658
Wohlfert, Beth
University at Buffalo
716-829-3969
Yang, Qi
Albany Medical College
518-264-2582
Yates, Jennifer
Wadsworth Center
518-339-2556
Yokoyama, Wayne
Washington University at St. Louis
314-229-8797