HIV Reservoirs in Visconti Patients and HIV Controllers

1. Kuala Lumpur, 2nd July 2013 HIV Reservoirs in Visconti Patients and HIV Controllers Pr Christine ROUZIOUX Virology, CHU Necker Université Paris Descartes

2. Productive infected Cells 2-LTR circles Total HIV DNA Integrated HIV-DNA Cell associated RNA US RNA and MS RNA Cellular Viremia Selected CD4+ T Cells Infectious (IUPM) HIV RNA + SCA Selected Resting CD4+ T Cells Infectious (IUPM) Resting infected Cells Lewin & Rouzioux, 2011

3. The ANRS assayfor HIV DNAquantification • Real time PCR (LTR gene, adapted to different HIV-1 subtypes • Sensitivity 3 copies/well of PCR • Same technique for more than 10 years • Reproducible and specific (HIV diagnostic in babies) • Commercialized by Biocentric (Bandol, France) • All Batches are controlledbefore release • All researchlabscan use the kits, sotheycan have comparable resultsusing the sameprocedures and the same standard for quantitation • Ultra-sensitive: 6 to 8 PCR to explore the wholeextract and a maximum of cells to catch the lowest values

4. Total cell-associated HIV-DNA in > 1500 patients included in the ANRS Cohorts: PRIMO, SEROCO, ALT, HIC and Visconti 6 5 4 AIDS HIV-DNA (log10 copies/106 PBMC) 3 2 1 PHI Chronic cART ALT HIC PTC Post-treatment controllers have an extremely low HIV-1 reservoir Lewin and Rouzioux, AIDS, 2012

5. ANRS : Long Term Asymptomatic Cohort 1000000 100000 HIV-1 RNA (log10 copies/ml) 10000 1000 100 10 1 10 100 1000 10000 100000 HIV-1 DNA (log10 copies/106 PBMC) HIC

6. HIV reservoir distribution in PBMC from Visconti Patients PBMC Live Resting (CD25-CD69-HLA DR-) CD4 Subsets Sorting (Flow Cytometry, ARIA SORP II in a L3 facility) Total HIV-DNA quantification • Ultrasensitive real-time PCR • Frequence of positive cells • distribution • subsets contribution • of/to the HIV reservoir In vitro induction of HIV replication from latent reservoirs 13-day long culture of each sorted-subsets • HIV-RNA quantification by RT-PCR • reservoir’s inducibility

7. PTCs have a low HIV reservoir distributed in the resting memory CD4 T cell subsets p = 0 . 0 0 1 p = 0 . 0 0 2 p = 0 . 0 0 1 p = 0 . 0 0 2 p = 0 . 0 0 9 8 p = 0 . 0 0 6 8 p = 0 . 0 0 5 9 p = 0 . 0 0 9 8 5 A ) 4 N s D l l - e V c I H 6 3 0 d 1 e / t s a e i i c p o 2 o s c s a g - o l l L e ( 1 C 0 C D 4 P B M C C D 4 C D 4 T N T C M T T M T E M M o n o c y t e s A C T + A C T - Saez-Cirion et al Plos Pathogens 2013

8. Weak contribution of long-lived resting CD4+ T cells to the HIV reservoir in the post-treatment controllers. Sáez-Cirión et al, ANRS VISCONTI Study. 2013

9. PHI Patients at PHI :ANRS OPTIPRIM trial at inclusion PTC

10. HIV replication is inducible from the resting memory CD4 T cell subsets from post-treatment controllers. Sáez-Cirión et al, ANRS VISCONTI Study. PLoS Pathog 9(3): e1003211. doi:10.1371/journal.ppat.1003211

11. Post-treatment controllers have very low levels of cell-associated HIV DNA which keep decreasing after treatment interruption for some patients. Sáez-Cirión et al, ANRS VISCONTI Study. 2013

12. Acknowledgements Patients and clinicians who participate to the cohort studies Institut Pasteur Régulation des Infections Rétrovirales Asier Saez-Cirion Gianfranco Pancino Daniel Scott-Algara Françoise Barré-Sinoussi Pierre Versmisse CHU Necker Enfants Malades Université Paris Descares Christine Rouzioux Véronique Avettand-Fenoel Adeline Mélard Antoine Chéret CHU Pitié-Salpetriere INSERM UMR-S 945 Brigitte Autran Charline Bacchus Benjamin Descours Assia Samri Ioannis Theodorou Julien Guergnon Faculté de Médecine Paris Sud INSERM U1012 Cécile Goujard Alain Venet Olivier Lambotte Isabelle Girault Jean-François Delfraissy INSERM U1018 Laurence Meyer Faroudy Boufassa CHR Orléans La Source Service Maladies Infectieuses Laurent Hocqueloux Thierry Prazuck All Funded by the CHU Hôtel-Dieu Université Paris Descatres Jean-Paul Viard Jade Ghosn ANRS CO6 ANRS CO15 ANRS CO18 Essai ANRS 147 PRIMO ALT HIV controllers OPTIPRIM