Charani RanasingheMolecular Mucosal Vaccine Immunology Group, Dept Immunology

The John Curtin School of Medical ResearchThe Australian National University

Novel HIV IL-4R antagonist vaccine strategy can induce both high avidity CD8 and excellent B cell immunity

2

Mucosal vaccination induces high quality/avidity HIV-specific CD8 T cells

Induction of high quality/avidity HIV-specific CD8 T cells following mucosal vaccination is associated with lower expression of IL-4/IL-13 by CD8 T cells

Ranasinghe et al. J. Immunol 2007

Ranasinghe et al. Euro J Immunol 2009

Ranasinghe et al. Mucosal Immunology 2013

Absence of IL-4/IL-13 induces high avidity HIV-specific CD8 T cells

Construction of poxviral vector-based vaccines that co-express IL-4R antagonist using homologous recombination

IL-4R antagonist HIV gag/pol/env

Recombinant vaccinia virus (rVV) or Modified Vaccinia Ankara (rMVA) - booster vaccine

Recombinant fowlpox virus (rFPV) - priming vaccine

HIV gag/polIL-4R antagonist

Jackson Boyle Ranasinghe Methods in Molecular Biology (2014)

intranasal/ intramuscular (i.n./i.m.) combined mucosal/systemic strategy

IL-4

Rα

γc IL-4

Rα

IL-1

3Rα

1

IL-13Rα2m

STAT6

IL-4 IL-13

4

Novel IL-4R antagonist adjuvanted vaccine will bind to IL-4R and transiently block IL-4/IL-13 signaling via the STAT6 pathway

Ranasinghe et al Cytokine and Growth Factor Reviews (In press)

96 hrs

12 hrs

Jackson Worley Trivedi Ranasinghe (submitted)

5Jackson Worley Trivedi Ranasinghe (submitted)

i.n./i.m. IL-4R antagonist adjuvanted vaccine strategy can induce HIV-specific CD8 T cells of high avidity

Inclusion of the inhibitor in the priming vaccination is crucial to induce the high avidity T cell repertoire

6.9% 9.8% 21.8% 19.6%

0.38% 1.16% 0.76% 0.98%

FPV HIVIL-4C118/VVHIVIL-4C118

Spleen

Genito-rectal nodes

FPV HIV/VV HIV FPV HIVIL-4C118/VV HIV

CD8

Kd

-Gag

Inclusion of the inhibitor in the i.n. rFPV priming vaccination is crucial to induce high avidity CD8 T cell repertoire

FPV HIV/VV HIVIL-4C118

Jackson Worley Trivedi Ranasinghe (submitted)

( i)

FP

V H

IV∆

10/ V

V H

IV∆

10(i

i) F

PV

HIV

VV

HIV

Spleen Iliac nodes Lung Lung nodes

Ranasinghe Mucosal Immunology 2013

Novel vaccines can enhance both systemic & mucosal HIV-specific poly-functional HIV-specific CD8 T cell immunity

8Jackson Worley Trivedi Ranasinghe (submitted)Ranasinghe et al Mucosal Immunology 2013;

i.n./i.m. IL-4R antagonist adjuvanted HIV vaccine strategy induces excellent CD8 T cell mediated protective immunity

* * *

P < 0.05 compared to control vaccination

* 0.0567

i.n./i.m. IL-4R antagonist adjuvanted vaccine strategy can induce Gag-specific antibody class switching

Statistics were calculated using Mann – Whitney U test

* 0.0256*** 0.0006* 0.0566

* 0.0256

6 weeks 12 weeks

IgG1

IgG2a

9Jackson Worley Trivedi Ranasinghe (submitted)

10Ranasinghe et al Mucosal Immunology 2013 ; Trivedi Jackson Ranasinghe (Submitted)

100 101 102 103 104

cd103 FITC-A

DC_1 fpv balbc.fcs

100 101 102 103 104

cd103 FITC-A

DC_1 fpv balbc.fcs

100 101 102 103 104

cd103 FITC-A

DC_3 fpv c118.fcs

100 101 102 103 104

cd103 FITC-A

DC_3 fpv c118.fcs

CD

11b

CD103

FPV-HIV - unadjuvanted FPV-HIV IL-4RC118 (IL-4R antagonist)

73.5%

1.03%

45.1%

1.92%

i.n. delivery of FPV-HIV IL-4R antagonist adjuvanted vaccine recruits unique antigen presenting cells to the lung mucosae responsible for

the induction of high avidity CD8 T and excellent B cell immunity

CD11C+ CD11b+ CD103-

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Unique features of the novel i.n./i.m. HIV IL-4R antagonist adjuvanted vaccine strategy.

Help recruit unique antigen presenting cell subsets to the lung mucosae

Induce enhanced high quality/avidity mucosal & systemic HIV Gag-specific CD8 T cell immunity*

HIV Gag-specific antibody class switching (IgG1 and IgG2a)* Env-specific IgG1 following a second i.m. Env protein booster**

Induce triple action immunity

The immune responses induced are consistent with • HIV controllers* and • Features of partial protective efficacy in the RV144 trial**

=> Platform technology against other chronic pathogens

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Acknowledgements

Molecular Mucosal Vaccine Immunology Group:Ronald JacksonAnnette BuchananLisa Pavlinovic, Megan Glidden, Sherry Tu

Students: Danushka Wijsundara, Shubhanshi

Trivedi, Matthew Worley

ANU/BRF - Kerong Zhang

JCSMR/MCRF - Harpreet Vohra, Mick Devoy

ANU/Animal services staffCollaborators:

David Boyle - CSIRO AAHL

John Stambas - Deakin Uni/ CSIRO AAHL

Robert Center - Burnet Institute/ Uni of Melbourne