Pr. Patrice DEBRE Laboratoire Immunité & Infection INSERM UMR-S 945 Hôpital Pitié-Salpêtrière

1. “Towards an HIV Cure” Pre-Conference Symposium 20 & 21 July 2012 Towards a therapeutic vaccine to reduce HIV reservoir, both neutralizing HIV and inhibiting effector T cells depletion Pr. Patrice DEBRE Laboratoire Immunité & Infection INSERM UMR-S 945 Hôpital Pitié-Salpêtrière Paris, France

2. A strategy to decrease HIV reservoir Virus Neutralization Vaccine INFECTION Restore Immune-response PATHOGENESIS Non-infected CD4 cells Infected CD4 cells Virus NK

3. An epitope based vaccine, both 1) Inhibiting the deleterious effect of NK cells A NK ligand on CD4+ T cells ? 2) Neutralizing HIV-1

4. 40 30 UT 3S 20 CD4 10 2.7% 17.4% 0 NKp44L HIV-1 gp41 Env Fusion Peptide HR1 HR2 TM 517 532 643 678 558 595 3S motif NKp44L is induced during HIV infection by the 3S/gp41 motif Kill % NKp44L CD4 4+ 3- 8+ 8+ 4+ 3- Control PBMC HIV+ PBMC NKp44L 40 MHC-I p=0.002 30 NKp44 % NKp44L - + 20 NK 10 0 0 250 500 1000 750 3S : very conserved motif & specific to HIV-1 CD4 / mm3 Vieillard et al Proc Natl Acad Sci USA 2005

5. gC1qR is the cell-surface receptor of the 3S/gp41 motif on CD4+ T cells 3S IgM NA HK C1q 3S C3 3S+ agC1qR NA 0 25 50 75 100 125 150 175 NKp44L expression NKp44L expression (MFI) Listeria monocytogenes Staphylococcus aureus Plasmodium falciparum HCV, rubella virus, HSV and HTNV Deleterious effect of NK cells ? Role of gC1qR ! Fausther-Bovendo et al PLoS Pathogens 2010

6. Anti-3S Ab : a predictive value for the evolution of the disease ? 300 200 Anti-3S (U/mL) Multivariate studybetween 40 patients (first quartile) with a slope CD4>2.7/month and 40 patients (last quartile) with slope CD4<-6.9/month 100 IC95 Variable Odds-ratio P 0 Sexe M W Age (10y) CD4 (X100) Log VL 3S Ab(x100) 1.00 1.83 1.30 0.58 1.17 2.89 0.049-6.87 0.76-2.23 0.42-0.81 0.55-2.51 1.60-5.17 0.372 0.334 0.001 0.682 <0.001 0 400 800 1200 CD4 count/mm3 Vieillard et al AIDS (2006) Unpublished data

7. Summary-1 • NKp44L is specifically expressed on non infected CD4+ T cells from HIV-1 patients; • NKp44L is induced by a specific peptide from the gp41 HIV-1 protein; • Anti-3S antibodies are predictive of the disease evolution.

8. A 3S vaccine inhibiting HIV pathogenesis Proof of concept in macaques 1- Macaque model of SHIV infection (Vieillard et al, AIDS 2008) 2- Prophylactic vaccination (Vieillard et al, PNAS 2008) 3- Therapeutic vaccination (Vieillard et al, submitted) Collaboration: Roger Le Grand (CEA, Fontenay-aux-roses; France)

9. Prophylactic vaccination by the 3S-gp41 peptide Study design Infection SHIV162P3 Immu 1 Imm. 2 Imm. 3 Sacrifice D0 D30 D60 D385 D585 • Analysis: • Anti-3S antibodies • Plasma viral load • Blood lymphocyte phenotyping • NK & CD4 activation • Cytotoxicity • Apoptose • Inflammation Animal groups : KLH-control macaques : 3S-immunized macaques : Vieillard et al Proc Natl Acad Sci USA (2008)

10. SHIV162P3 Effect of 3S vaccination on Viral load, NKp44L expression, and CD4 count 3S 3S 3S 1500 NS NS 3S 3S 108 anti-3S (U/mL) 106 1000 3S Viral load 104 3S 500 102 0 1 -400 200 -200 0 0 50 100 150 200 50 2000 ** * 40 % CD4+NKp44L+ 1500 CD4 / mm3 30 1000 20 500 10 0 0 0 50 100 150 200 0 50 100 150 200 Days post-infection KLH-control macaques 3S-immunized macaques

11. 10 7.5 5 2.5 0 Effect of 3S vaccination on CD4 activation & Inflammation % CD4+CD69+ 0 20 40 60 80 Days post-infection 80 50 * 40 60 ** CRP (mg/ml) ** TNF-(pg/ml) 30 40 20 20 10 0 7 21 0 0 21 0 7 Days post-infection 3S-immunized macaques KLH-control macaques

12. Summary-2 • Immunogenicity in cynomolgus (anti 3S response) • BUT Unchanged viral load • Decreased NKp44L expression on CD4+ T cells • Decreased NK cells cytotoxicity on CD4+ T cells • Decreased CD4+ T cells apoptosis • Preservation of CD4+ T cell counts • Decreased immune activation • Decreased inflammation during the acute phase

13. Summary 40 44L+ 30 % NK Lysis 20 44L- Autologous NK cells HIV infected cells 10 0 100/1 50/1 25/1 12.5/1 E/T ratio HIV virion NKp44 NKp44L gp41 Epitope 3S (SWSNKS) CD4+ T cell lysis CD4+NKp44L+ cell CD4+ T cell CD4depletion Anti-3S Ab Cell activation Inflammation

14. An epitope based vaccine, both 1) Inhibiting the deleterious effect of NK cells 2) Neutralizing HIV-1 Search for an effect of 3S mutant NH2-SWSNKS-NH2 Alanine scanning

15. HIV infectivity of 3S mutants Infectivity 100 75 % p24 production 50 25 0 WT S615A S618A W614A N616A K617A S613A Entry 100 75 % b-galactosidase activity 50 25 0 WT S615A S618A W614A N616A K617A S613A

16. Neutralizing effect of anti-3S mutant Abs Human Abs Mice Abs 200 200 WT 150 150 200 p24 (ng/ml) p24 (ng/ml) WT 100 100 150 100 50 50 50 WA #109 0 0 5 10 15 20 0 0 0 5 10 15 20  Ig (mg/ml)  Ig (mg/ml) W/o Ab 200 WT WT 150 p24 (ng/ml) 100 50 #109 WA 0 4 0 2 6 8 10 4 2 6 0 8 10 Days post-infection Days post-infection

17. Antibodies against 3S (WA) mutants also inhibit NK pathogenesis WT 3S NT 68.3 6.4 #44 #117 #24 WT 3S NT 0.6 2.9 7.9 28.6 21.9 34.6 28.4 1.6 2.1 32.1 64.2 58.2 38.3 34.1 29.4 #65 #109 #71 #44 #24 #117 22.7 21.9 31.3 1.4 7.7 2.1 4.9 1.8 1.8 CD107a 61.0 29.9 52.1 40.9 55.1 30.3 NKp44L #109 #65 #71 2.3 6.3 1.9 7.8 1.1 5.9 55.0 36.4 55.4 34.9 62.6 30.4 NKp44

18. Conclusion A gp41 epitope based vaccine, both 1) Restoring/preserving CD4 cell functions 2) Neutralizing HIV-1 As a tool to decrease HIV reservoir

19. In Progress … Clinical trial Phase I/IIa Animal Model Therapeutic Vaccine Non-infected Asymptomatic phase AIDS HIV-1 infection Antiretroviral Therapy

20. Partners Hôpital Pitié-Salpêtrière - Paris CEA Unité Immunité & Infection Team-2 Patrice Debré Hugues Fausther Bovendo Caroline Petitdemange Alexis Sennepin Abla Achour Florence Baychelier Vincent Vieillard Team-1 Brigitte Autran Assia Samri Unité Epidémiologie Clinique Dominique Costagliola Service de Virologie Henri Agut Vincent Calvez Daniel Candotti Isabelle Malet Service des Maladies Infectieuses & Tropicales Christine Katlama Laboratoire d’ Immuno-Virologie Roger Le Grand Nathalie Deudreude-Bosquet Aurélien Corneau Isabelle Mangeot-Méderlé … Grants ANRS Sanofi-Pasteur Bill & Melinda Gates Foundation Agence Nationale de Recherche (ANR) InnaVirVax ORVACS Institut Pasteur Olivier Schwartz Nathalie Sol-Foulion Felix Rey