T cell Dependent B cell Activation Plays a Role in Mediating

Hypertension and Pathophysiology in response to CD4+ T cells from

Reduced Uterine Perfusion Pressure (RUPP) in Pregnant Rats

Council for High Blood Pressure ResearchOral III: Top Trainee Oral Abstracts

September 10, 2014Denise C. Cornelius, Ph.D.

University of Mississippi Medical Center, Jackson, MS

DISCLOSURES

NONE

Pathophysiology of Preeclampsia

Adapted: Parham PJ, Exp Med 2004, Parikh SM & Karumanchi SA. Nature Medicine 2008 14, 810 - 812

Inflammatory cytokines and ET-1

B-Cell Activation

With T (cells) ? or Not with T (cells)?Adapted from Dörner, et al. 2009. Nature Reviews Rheumatology 5, 433-441.

HYPERTENSION

Reactive Oxygen Species

Endothelin-1

Placental Ischemia

CD4 + TRegIL-10

ActivatedCD4 + THelper

Cells

B cell stimulationAT1-AA

LaMarca, Cornelius, Wallace. Physiology (Bethesda). 2013;28(4):225-33. ISSN: 1548-9221

Reduced Uterine Perfusion Pressure

Pregnancy-induced Hypertension Characteristics

Hypertension + +

Proteinuria + +/-

Increase TNF alpha and IL-6 + +

CD4+ T cells and TRegs + +

Endothelial dysfunction + +

AT1 and ETA autoantibodies + +

HUMAN RUPP

Hypothesis

T cell Dependent B-Cell Activation

Shekhar and Yang.2012. Cellular& Molecular Immunology. 9: 380-385

• CD40 Ligand – T cells• CD40 – B Cells• Interaction causes

• Activation/differentiation of B-cells• Inflammation

Methods: Cross-Talk Inhibition

MAP taken and organs harvestedOvernight Culture with IL-2,

IL-12, & αCD40 Ligand

CD4+ T cell Isolation from RUPP

spleen

Inject 1x106 cellsinto GD12 NP

Culture Day 1 Culture Day 2

FACS analysis

CD4+

CD4+ T cells + αCD40-LIL-2

IL-12

Insertion of carotid catheters on GD18

G Day 18 G Day 19

CD4+ T cells

Blockade of CD40 ligand on RUPP CD4+ T cellsblunts chronic inflammation associated with

adoptive transfer of RUPP CD4+ T cells

NP

NP +

RP T

c e ll

NP +

RP C

D4 0L

pg

/mL

NP

NP +

RP T

c e ll

NP +

RP C

D4 0L

pg

/mL

n = 4n = 3

n = 4n = 4

n = 4

n = 4

Blockade of CD40 ligand on RUPP CD4+ T cellsinhibits oxidative stress associated with adoptive

transfer of RUPP CD4+ T cells

NP

NP +

RP T

c e ll

NP +

RP C

D4 0L

RL

U/m

in/m

g p

rote

in

n = 4

n = 4

n = 4

Blockade of CD40 ligand on RUPP CD4+ T cellsminimizes AT1-AA production associated with

adoptive transfer of RUPP CD4+ T cells

NP

NP +

RP T

c e ll

NP +

RP C

D4 0L

bea

ts/m

in (

bp

m)

n = 3

n = 3

n = 3

Blockade of CD40 ligand on RUPP CD4+ T cellsblunts hypertension associated with adoptive

transfer of RUPP CD4+ T cells

NP

NP +

RP T

c e ll

NP +

RP C

D4 0L

mm

HG

n = 18

n = 5

n = 21

Summary

HYPERTENSION

Reactive Oxygen Species

Endothelin-1

Placental Ischemia

CD4 + TRegIL-10

ActivatedCD4 + THelper

Cells

B cellsAT1-AA

Conclusion

XInhibit T-cell dependent B-Cell Activation (αCD40L)

Perspective & Clinical Relevance

• Proper immune function is important to inhibit, at least in part, the blood pressure response and pathophysiology associated with placental ischemia.• CD4+ T cells mediate some of the pathophysiology in PE

through B cell activation, and are a potential therapeutic target • Administration of antibody against CD40 ligand may

be a therapeutic option for management/treatment of preeclampsia.

AcknowledgmentsPharmacology

• Rich Roman Ph.D.• Babbette LaMarca, Ph.D.

• Janae Moseley• Lorena Amaral, Ph.D.

• Ashlyn Harmon• Justin Porter• Jeremy Scott

• Nathan Campbell• Alexia Thomas

Cardiovascular & Renal Research Center

• Joey Granger, Ph.D.Immunology

• Eva Bengten,Ph.D. • Melanie Wilson,Ph.D.

Obstetrics and Gynecology• Kedra Wallace, Ph.D.• Javier Castillo, M.D.

National Institutes of Health• RO1HD067541 • T32HL105324