T-cell responses induced by immunization with p yAd35-vectored HIV-1 vaccines are broad, durable, polyfunctional and can mediate inhibition of HIV

Josephine H Cox, PhD

AIDS Vaccine 2011Bangkok, Thailand, September 12-15th, 2011

Ad35-GRIN/ENV Phase I OverviewTwo constructs: Ad35-GRIN & Ad35-ENV

Ad35-GRIN/ENV: Groups A-C co-formulated together and Ad35-GRIN: Group Dtogether and Ad35 GRIN: Group D

All administered via intramuscular route at 0 and 6 months.

Subtype A

N = 56, 14/group: 10 vaccine/4 placebo

A (Low) B (Mid) C (High) D (Mid)

Ad35 GRIN/ENVAd35-GRIN/ENVDose escalation

2x109 vp 2x1010vp 2x1011 vp

Ad35-GRIN 1x1010 vp

Clinical Site: University of Rochester Medical Center (USA)Vaccinations completed Sept. 2010Follow up for 18 months post first vaccinationFollow-up for 18 months post first vaccination

Last visit of last volunteer August 20112

Safety Data

• Only Ad35 antibody negative subjects were enrolled. • Ad35 prevalence of 4.9% low in this US populationAd35 prevalence of 4.9% low in this US population

• Ad35-GRIN/ENV is safe and generally well tolerated- No related SAE reported - Reactogenicity and tolerability are dose-dependent- High dose at 2x1011 vp is responsible for severe local and/or

systemic reactions in 50% of vaccinees persisting more than 5systemic reactions in 50% of vaccinees, persisting more than 5 days in one volunteer.

- All reactions resolved spontaneously without complications or sequellaesequellae

• All Ad35 samples (scheduled and symptomatic) tested negative for shedding (urine and saliva).g g ( )

3

Immunogenicity Assessments

3535 Weeks0 24

Ad3

Ad3 Weeks

0, 2 and 4 post 1st 0, 1, 2, 4, 8, 14, 28, 40 and 48 post 2nd

•Immunogenicity: IFN- ELISPOT at all time points

Followed for 72 weeks total (18 months)

g y p•Cut-off 38 SFC/106 PBMC and 4 x background / six peptide pools

•ICS, Viral Inhibition Assay (VIA), epitope mapping, Env Ab, p24 Ab, Ad35 neuts at selected time pointsb, p b, d35 euts at se ected t e po ts

4

High HIV-specific IFN- ELISPOT Response rates maintained over time

60%

80%

100%

A Low

p

Any Antigen

0%

20%

40%

A-LowB-MidC-HighD-GRIN

0%

E A

60%

80%

100% Env Antigen

60%

80%

100%

20%

40%

60%

GRIN Antigen20%

40%

60%

0% 0%

5

HIV-specific IFN- ELISPOT in most individuals maintained up to 10 months post 2nd vaccination p p

3.5Ad35 GRIN-ENV increasing dose Ad35 GRIN

3.0

scal

e)

2.5

BM

C (l

og-s

2.0

FC/1

06PB

1.5

Wk 250%Vac.1

Wk 450%

Wk 286%Vac.2

Wk 475%

Wk 4063%

Wk 250%Vac.1

Wk 456%

Wk 2100%Vac.2

Wk 4100%

Wk 40100%

Wk 278%Vac.1

Wk 470%

Wk 289%Vac.2

Wk 488%

Wk 4075%

Wk 289%Vac.1

Wk 490%

Wk 286%Vac.2

Wk 486%

Wk 4088%

SF

38 SFC

2x109 vp 2x1010 vp 2x1011 vp 1x1010 vp6

IFN- ELISPOT responses of modest magnitude across all vaccine-expressed HIV proteins

35Gag RT Pol/Int Nef Env

3.0

3.5

og-s

cale

)

2.5

PBM

C (l

o

2.0

SFC

/106

1.5

Grp A43%

Grp B88%

Grp C33%

Grp D86%

Grp A57%

Grp B88%

Grp C56%

Grp D71%

Grp A71%

Grp B38%

Grp C33%

Grp D71%

Grp A29%

Grp B38%

Grp C11%

Grp D57%

Grp A86%

Grp B75%

Grp C67%

Median SFC/m 158 79 100 158 126 251 158 200 126 79 251 316 158 50 79 100 63 126 79

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2 weeks post second vaccination

Multiple HIV proteins targeted by each volunteer

Number of visits (maximum 10) with

Breadth of the response as defined by number of positive

responses to any of the sixpositive responses to

any peptide pool

responses to any of the six (four for Group D) peptide

pools at any time point

Gro p MedianGroup Median

A - Low 7.5 3.5

B - Mid 9 5

C - High 9 5C - High 9 5

D - GRIN 10 4

Data up to 40 weeks post second vaccination8

Median of 4 epitopes recognized per volunteer (n = 25 mapped)volunteer (n = 25 mapped)

Pool ID#

VolunteersUnique

*regionsMedian # *regions RangePool ID Volunteers regions

mappedregions

recognizedRange

Gag 12 >8 1 1-3RT 14 17 2 1 3RT 14 >17 2 1-3Int 12 >8 1 1-4Nef 6 >2 1 1-2Env 12 >14 2 1-4

Median # epitopes recognized / volunteer 4Range of epitopes recognized / volunteer 1-8

9*Unique region = either 2 peptides sharing a sequence or single peptide

Flow Cytometry Panel: IFN-/IL-2/CD107/TNF-, CD4/CD8 memory and homing markersCD4/CD8, memory and homing markers

CD4+: 0.028% IFNg+CD8+: 2.18% IFNg+ NEF ELISPOT: 1060 SFC/m

Example shown is Nef stimulation from sample atExample shown is Nef stimulation from sample at 2 weeks post 2nd vaccination (Group D, GRIN)

10

Predominantly CD8+ T-cell Responses to Gag, RT and Int

>0.5Any Env 1 Env 2 Gag INT NEF RT

CD8 ANY Env 1 Env 2 Gag Int Nef RT

/TN

F

0.2

0.3

0.4

-2/C

D10

7/

Baseline & Placebo

0.13

0.0

0.1

Bl P B C D B C D B C D B C D B C D B C D

Any Env1 Env2 Gag INT NEF RTe IF

N-

/IL-

B - Mid

C - HighCD4 ANY Env 1 Env 2 Gag Int Nef RT

0.3

0.4

>0.5Any Env 1 Env 2 Gag INT NEF RT

e Po

sitiv

e C High

D - GRIN

0.06

0.1

0.2

0.3

% C

ytok

ine

11

0.06

0.0Bl P B C D B C D B C D B C D B C D B C D

%

Responses at 2 weeks post 2nd vaccine

Potent and broad CD8-mediated VIA activity Placebo Low dose    Mid dose    High dose    

CRFAE-01 ELI (AD) IIIB(B) NL4-3(B) U455(A)

inhi

bitio

nLo

g 10

247Fv2(C) 97ZA012(C) CBL4(D) CH077(B) CH106(B)

CD8+ T cell HIV-1 inhibition to several isolates in most vaccinated individuals 12

ENV-A (UG037)- and Gag-p24 (IIIB)-specific ELISA antibodiesspecific ELISA antibodies

10 4Post 1st Vaccination Post 2nd Vaccination

rs

10 4Post 1st Vaccination Post 2nd Vaccination

rs

10 3

A - LowB - MidC - High

an E

nv ti

ter

10 3

A - LowB - MidC - HighD - GRINan

Gag

tite

r

10 2Geo

m M

ea

10 2Geo

m M

ea

0 10 20 30 4010

Weeks post Vaccination0 10 20 30 40

10

Weeks post Vaccination

4 w Post 1st 2 w Post 2ndA - Low 8/10 (80) 9/9 (100)B - Mid 9/10 (90) 8/8 (100)

4 w Post 1st 2 w Post 2ndA - Low 0/10 0/9 (0)B - Mid 0/10 3/8 (38)C High 0/10 8/9 (89)

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C - High 10/10 (100) 9/9 (100) C - High 0/10 8/9 (89)D - GRIN 0/10 5/7 (71)

T-cell response summary for B001 Ad35 trialHigh response rate by ELISPOT >86% across all groups post 2nd

Magnitude of response is modest but maintained over time

Greater magnitude to GRIN proteins in the absence ofGreater magnitude to GRIN proteins in the absence of ENV but no enhancement of VIA activity

Good breadth; all proteins targeted CD8+ T-cell HIV-1 inhibition in most vaccinated individuals

to several isolatesFl t tFlow cytometryPredominantly CD8+ response Multifunctional (cytokines + degranulation)Multifunctional (cytokines + degranulation)Induction of T cell memory responses (data not

shown) 14

Antibody response summary for B001 Ad35 trialAnti-HIV ENV: 100% responders post second

administration with modest titersNo HIV Nabs to multi-clade panel (Monogram BioSciencesNo HIV Nabs to multi clade panel (Monogram BioSciences,

data not shown)

Anti-HIV Gag p24: no antibody post 1st, modest titer post 2ndpost 2nd

Anti-Ad35 neutralization response rates and titers are modest even after 2 vaccinations and lower than seen with Ad5 (data not shown)

Low-prevalence serotype Ad vectors may exhibit i d f t d i i it dimproved safety and immunogenicity as compared with Ad5 vectors.

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Next steps Ad35-GRIN/ENV is immunogenic at the dose with an acceptable safety

profile (2x1010 vp) Evaluate the safety and immunogenicity of Ad35-based HIV vaccine in

heterologous prime-boost regimens, to assess potential enhancement of the breadth and potency of cellular and humoral immune responses as follows:

Ad26 vector (Dan Barouch / Crucell, ongoing)Ad35 ENV d Ad26 ENV P t tAd35-ENV and Ad26-ENV PrototypeMosaic Immunogens to optimize cellular immunologic coverage of global HIV-1 sequence diversity

Adjuvanted Protein (GSK ongoing)Adjuvanted Protein (GSK, ongoing)Ad35-GRIN and Adjuvanted GSK investigational HIV vaccine formulation 1 and 2

Electroporated (EP) DNA and IL-12 (Profectus start late 2011)Electroporated (EP) DNA and IL 12 (Profectus, start late 2011)Ad35-GRIN-Env and multi-antigenic DNA (HIV-MAG), IL-12 and EP

Mucosal delivery – Sendai virus (SeV)Ad35-GRIN and SeV-Gag prototype in pre-clinical developmentAd35 GRIN and SeV Gag prototype in pre clinical development

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AcknowledgementsUniversity of RochesterUniversity of RochesterMedical CenterMichael KeeferLottie HachaambwaCatherine Bunce

IAVIEddy Sayeed, Angela Lombardo Wendy Komaroff, Sabrina Welsh

Catherine Bunce Mhorag HayEMMESKelley LoughranBurc Barin

James Ackland, Devika ZachariahKamaal Anas, Jean-Louis Excler Patricia Fast

Burc BarinCarol Smith

IAVI Human Immunology Laboratory, Imperial College

Lorna ClarkJustyna Czyzewska

Laura SeamonsAggeliki SpentzouLaboratory, Imperial College

LondonHong WanAmbreen AshrafPhil Bergin

Justyna CzyzewskaChristopher FarranceNatalia FernandezDilbinder GillP t H

Aggeliki SpentzouTony TarragonaParamesh ChettyGwynn StevensJill GilPhil Bergin

Emmanuel CormierJana CargaMichelle Cashin-CoxHannah Cheeseman

Peter HayesVanaja KakarlaJakub KopycinskiFrancesco Lala

Jill Gilmour

Hannah Cheeseman Andrew Raxworthy-Cooper

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Back- up slide

Range of CD8+ T-cell memory and other phenotypes observedphenotypes observed

No trend towards a particular memory phenotype. Majority of responses lack homing markers

d B7Data is across study arms and visits in samples with >0.2% IFN- production

and are B7-

Ad35-specific neutralization antibody responses

100 500

60

80

resp

onde

rs

300

400

GM

T

0

20

40

% A

d35

0

100

200

0 1 2 0 1 2 0 1 2 0 1 2 0 1 2 0 1 2 0 1 2 0 1 20

D - GRINA - Low B - Mid C - High

0 = baseline1 4 k t fi t i1 = 4 weeks post first vaccine2 = 2 weeks post second vaccine