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and HIV Controllers Kuala Lumpur, 2nd July 2013

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1 and HIV Controllers 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 Cellular Viremia Resting 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 Fisrt of all I would like to address the key poit which is still matter of debate How to measure HIV reservoirs On this slide the different methodologies that may be used Quantification of nucleic acid technique Or culture to show the capacity of cells to produce infectious virus We have developped a quantitative real time PCR Selected Resting CD4+ T Cells Infectious (IUPM) Resting infected Cells Lewin & Rouzioux, 2011

3 The ANRS assay for HIV DNA quantification
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 controlled before release All research labs can use the kits, so they can have comparable results using the same procedures and the same standard for quantitation Ultra-sensitive: 6 to 8 PCR to explore the whole extract and a maximum of cells to catch the lowest values

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

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

6 HIV reservoir distribution in PBMC from Visconti Patients
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 1 2 3 4 5 P B M C D o n c y t e s A T + - N E p = . 9 l a i d H I V ( L g / 6 ) 8 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): e doi: /journal.ppat

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 CO ANRS CO Essai ANRS 147 PRIMO ALT HIV controllers OPTIPRIM 12

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