1. Contribution of the immune system to HIV persistence Sharon R Lewin Director, Infectious Disease Unit, Alfred Hospital Professor, Department of Medicine, Monash University Co-head, Centre for Virology, Burnet Institute, Melbourne, Australia Towards an HIV Cure, 5 th IAS Conference, Rome, 2011

2. Outline  Establishing HIV latency –Where and how are resting T-cells infected? –Chemokines –Dendritic cells  Maintaining HIV latency –Homeostatic proliferation –Inflammation  Role of the adaptive immune response  Relevance to strategies for cure

3. establishing HIV latency

4. Latent infection can be established in many T cells ThymusPeripheral circulation MM M M Effector/ transitional Central memory Ag Central memory Bone marrow Chun et al., Nature 1997; 387:183; Finzi et al., Science 1997; 278:1295; Brooks et al., Nat Med 2001; 7:459 ; Chomont et al., Nat Med 2009; 15: 893; Dai et al., J Virol 2009: 83(9):4528-37; Carter et al., Nat Med 2010; 16: 446; Wightman et al., J Infect Dis 2010; 202(11):1738-48 Naïve T cellsTransitional memory Multipotent progenitor cells Latent reservoir Naive

5. The main reservoir is central and transitional memory T-cells Chomont et al., Nature Med 2009;15: 893

6. GI tract is enriched for latently infected cells Chun et al., J Infect Dis 2008; 197:714; Yukl et al., J Infect Dis 2010; 202(10):1553-6 N=8, time on HAART with undetectable HIV RNA 2.8 – 12 years HIV DNA Copies per million cells

7. Most latently infected and productively infected cells are in lymphoid tissue North et al., J Virol 2010; 84(6):2913-22 Mmu 35389 VL=17 CSF VL<50 DNARNA Lymphoid tissue ++++++ GI tract+++ CNSNeg/+Neg Reproductive tissue +Neg Other+Neg RT-SHIV infection HAART =Tenofovir/emtricitabine/efavirenz

8. Size of HIV Reservoir correlates with activated CD8+ T cells (in Sigmoid Colon) Seth et al., Mucosal Immunology 2008:1:382-388

9. What is unique about these tissue sites?  Ongoing replication due to –Poor penetration of drugs? –Localised sites of inflammation?  Unique long-lived cells such as macrophages or DCs allowing for cell-cell transmission?  Infection of resting cells?

10. Unactivated resting cells Resting CD4+ T-cell Ex vivo tissue blocks Eckstein et al, Immunity 2001; 15: 671; Kreisberg et al., J Exp Med 2006; 203:865; Infection of resting T-cells in vitro chemokines In vitro Saleh et al., Blood 2007; 110:416; Cameron et al., Proc Natl Acad Sci 2010 epub Sept 18

11. Multiple chemokines can induce latent HIV infection in resting CD4+ T cells. Cameron et al., Proc Natl Acad Sci 2010; 107(39):16934-9

12. Chemokine receptor ligation activates cofilin and actin polymerisation CXCR4 + gp120 CCR7 + CCL19 Yoder et al Cell 2008 Cameron et al PNAS 2010

13. Chemokine signalling pathways PI3K PI3K

14. Inhibition of Erk1/2, Jnk and NF-kB eliminates integration (ALu-LTR) SC-514 Bay 11-7082 SP600125 SB203580 PD980509 CCL19+DMSO JNKERK NF  B P38 Saleh et al., 5 th IAS Conference, Rome, 2011

15. Unactivated resting cells Resting CD4+ T-cell Ex vivo tissue blocks Eckstein et al, Immunity 2001; 15: 671; Kreisberg et al., J Exp Med 2006; 203:865; Infection of resting T-cells in vitro chemokines In vitro Saleh et al., Blood 2007; 110:416; Cameron et al., Proc Natl Acad Sci 2010 epub Sept 18 Dendritic cells

16. Dendritic cells facilitate latent HIV infection Evans et al., unpublished

17. maintenance of HIV latency

18. What is the fate of a latently infected cell? Negative regulators differentiation EM activation Homeostatic proliferation

19. Number of latently infected cells is correlated with proliferation: role of IL-7 Chomont et al., Nat Med 2009

20. IL-7 increases proliferation leading to expansion of HIV DNA in vivo HIV infected subjects under suppressive HAART received IL-7 injections (ACTG5214, blind study). Vandergreeten 6 th IAS conference on HIV Pathogenesis, Treatment and Prevention, Rome, 2011 Day 0Day 28 0 200 400 600 800 p=0.03 Int HIV DNA per mL of blood Day 0Day 28 0 500 1000 1500 2000 Int HIV DNA per 10 6 CD4 T cells

21. What is the fate of a latently infected cell? Negative regulators differentiation EM activation Homeostatic proliferation

22. PD-1 hi cells are enriched for latentlly infected cells Da Fonesca et al., HIV Reservoir Workshop, Vienna, July 2010 Donor ADonor BDonor C Mock a-PD-1 blocking Ab Isotype control 200 400 600 0 p24 (pg/mL) Chomont et al., Nature Med 2009;15: 893

23. What is the fate of a latently infected cell? Negative regulators differentiation EM activation Homeostatic proliferation IL-15

24. role of the adaptive immune response

25. Inverse relationship between HIV- specific CD4+ T-cells and HIV DNA Martinez et al., J Infect Dis 2001; 191:2053–63 N=66; long term non progressors, no ARV

26. HLA type influences size and distribution of latently infected cells Descours; Avettand-Fenoel Submitted

27. Summary  Latency can be established in multiple T-cell subsets  High concentration of latently infected cells in tissue including GI tract and lymphoid tissue  Chemokines and dendritic cells play a key role in establishing latency in resting T-cells  Maintenance of latency can occur by –Homeostatic proliferation (IL-7) –Negative regulators of T-cell activation (PD1)  Adaptive immune response likely important

28. What is the fate of a latently infected cell? Negative regulators differentiation EM activation Homeostatic proliferation Anti-IL7Anti-PD-1 Auranofin IL-15 Chemokine antagonists vaccination minocycline

29. Acknowledgements  Department of Medicine, Monash University, Melbourne –Paul Cameron –Suha Saleh –Vanessa Evans –Nitasha Kumar –Ajantha Solomon –Georgina Sallman –Fiona Wightman  Westmead Millenium Research Institute, Sydney –Tony Cunningham –Andrew Harman  VGTI Florida, Port St Lucie, FL –Rafick Sekaly –Elias Haddad –Genevieve Boucher –Nicolas Chomont  Hopital Pitie Salpatriere, Paris –Brigitte Autran –Benjamin Descours