1. CAROL VINTON LABORATORY OF DR. JASON BRENCHLEY LABORATORY OF MOLECULAR MICROBIOLOGY NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES NATIONAL INSTITUTES OF HEALTH JULY 25 TH, 2012 AIDS 2012, HIV/SIV PATHOGENESIS SESSION Differential SIV infection patterns of lymph node-resident CD4 T cells distinguishes progressive from non-progressive SIV infection

2. Sooty mangabey SIVcpz Chimpanzee HIV-1 SIV SIVsyk Sykes monkey SIVcol Mantled guereza L-Hoest’s monkey Mandrill SIVlho SIVrcm SIVgsn SIVmnd Red-capped mangabey SIVagm African green monkey SIVsmm Greater spot-nosed monkey Slide courtesy of Beatrice Hahn HIV-2 SIVmac

3. Natural Host (SM/AGM/etc) Non-Pathogenic Infection Non-Natural Host (RM/PTM/humans) Pathogenic Infection YesHigh Viral LoadYes RareDepletion of Peripheral CD4+Yes SomeAcute Depletion of Mucosal CD4+Yes NoProgressive Depletion of Mucosal CD4+Yes NoPreferential depletion of Th17Yes NoMicrobial TranslocationYes NoImmune ActivationYes NoProgression to AIDSYes vs.

4. Nature Med. 2011 Jun 26;17(7):830-6. doi:10.1038/nm.2395. Low levels of SIV infection in sooty mangabey central memory CD 4+ T cells are associated with limited CCR5 expression. Paiardini M, Cervasi B, Reyes-Avilles E, Micci L, Ortiz AM, Chahroudi A, Vinton C, Gordon SN, Bosinger SE, Francella N, Hallberg PL, Cramer E, Schlub T, Chan ML, Riddick NE, Collman RG, Apetrei C, Pandrea I, Else J, Munch J, Kirchhoff F, Davenport MP, Brenchley JM, Silvestri G. Yerkes National Primate Research Center, Emory Vaccine Center and Department of Pathology, Emory University, Atlanta, Georgia, USA. Mirko.paiardini@emory.edu Are there immunological mechanisms associated with non-progressive SIV infection in natural hosts? 4+ Nature Med. 2009 Aug;15(8):879-85. CD4 downregulation by memory CD 4+ T cells in vivo renders African green monkeys resistant to progressive SIVagm infection. Beaumier CM, Harris LD, Goldstein S, Klatt NR, Whitted S, McGinty J, Apetrei C, Pandrea I, Hirsch VM, Brenchley JM. Lab of Molecular Microbiology, National Institute of Allergy and Infectious Diseases, US National Institutes of Health, Bethesda, MD, USA.

5. Experimental Layout Flow cytometrically sorted and analyzed CD4+ T cell subsets from PBMC and LN samples SM (natural, non-progressive SIV hosts) RM (non-natural, progressive SIV hosts) Analyzed SIV infection frequencies by qPCR

6. CD4+ T cell subsets analyzed Naïve: mature, antigen inexperienced T cells, reside in blood and lymphoid tissue Effector Memory (EM): antigen experienced, reside in blood and effector tissues Central Memory (CM): antigen experienced, long-lived pool of precursors for additional memory and effector cells, reside in blood and lymphoid tissue T follicular helper (Tfh): antigen experienced, promote B cell proliferation and maturation, reside in lymphoid follicles

7. Analysis and sorting of CD4+ T cell subsets Lymphocytes Single Cells Live cellsCD3+ Live Lymphocytes CD4+ cells Memory CD4+ cells Naive CD4+ cells Non-Tfh cells CM EM Tfh cells

8. SIV infected cell frequencies: PBMC SIVsmmE543-infected RM SIVsmm-infected SM SIVmac239-infected RM

9. SIV infected cell frequencies: LN SIVsmmE543-infected RM SIVsmm-infected SM SIVmac239-infected RM

10. Why are Tfh cells more prone to SIV infection in RM?

12. SIV+ cells in the GC of lymph nodes Within follicles Extrafollicular T cell zone

13. Trapping of Virions by FDCs

14. Summary Different anatomical & cellular viral reservoirs in natural (SM) and non-natural (RM) hosts for SIV CM CD4+ T cells of RM are 2-10x more frequently infected than SM CM CD4+ T cells – both in PBMC and LN Frequency of SIV-infected CD4+ Tfh cells is higher in RMs compared to SMs LN SIV viral burden is greater in RM than SM Preferential infection of Tfh cells in RMs is possibly linked to increased viral trapping by FDCs Preferential infection of Tfh cells in RM may, in part, contribute to abnormal antibody responses observed during progressive SIV infection

15. Acknowledgements Lab of Molecular Microbiology, NIAID, NIH Jason Brenchley Molly Perkins Lauren Canary Nichole Klatt AIDS and Cancer Virus Program, Frederick National Laboratory Jake Estes Jeff Lifson Brian Tabb Pathology and Histotechnology Laboratory, Frederick National Laboratory Xing Pei Hao Lab of Molecular Microbiology, NAID, NIH Vanessa Hirsch NIH Animal Center Heather Cronise Joanna Swerczek Rick Herbert Division of Infectious Diseases, University of Colorado Denver Elizabeth Connick Emory University Mirko Pairidini Guido Silvestri

16. RM maintain similar overall frequencies of SIV infected naïve, EM, and CM CD4+ T cells as SIV-infected SM

17. Maintenance of memory Tfh CD4+ T cell subset in SIV+ RM