HIV Reservoirs in Visconti Patients and HIV Controllers

Kuala Lumpur, 2nd July 2013

Pr Christine ROUZIOUXVirology, CHU Necker Université Paris Descartes

Total HIV DNAIntegrated

HIV-DNA

Cell associated RNAUS RNA and MS RNA

HIV RNA + SCA

2-LTR circles

Cellular Viremia

Selected CD4+ T Cells

Infectious (IUPM) Selected

Resting CD4+ T Cells

Infectious (IUPM) Resting

infected Cells

Productive infected Cells

Lewin & Rouzioux, 2011

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

PHI Chronic cART ALT HIC PTC

HIV-

DNA

(log1

0 co

pies

/106 P

BMC)

1

2

3

4

5

6

Total cell-associated HIV-DNA in > 1500 patients included in the ANRS Cohorts: PRIMO, SEROCO, ALT, HIC and Visconti

Lewin and Rouzioux, AIDS, 2012

Post-treatment controllers have an extremely low HIV-1 reservoir

AIDS

HIV

-1 R

NA

(log

10 c

opie

s/m

l)

HIV-1 DNA (log10 copies/106 PBMC)

10

100

1000

10000

100000

1000000

1 10 100 1000 10000 100000

ANRS : Long Term Asymptomatic Cohort

HIC

Live Resting (CD25-CD69-HLA DR-) CD4 Subsets Sorting(Flow Cytometry, ARIA SORP II in

a L3 facility)

Total HIV-DNA quantificationUltrasensitive 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

PBMC

HIV reservoir distribution in PBMC from Visconti Patients

0

1

2

3

4

5

P B M C C D4 M o n o cytes C D4A C T +

C D4A C T -

T N T C M T T M T E M

p = 0 .0 0 5 9

Cell -

asso

ciat

ed H

IV- D

NA(L

og c

opie

s/10

6 cel

ls)

p = 0 .0 0 9 8

p = 0 .0 0 9 8 p = 0 .0 0 6 8

p = 0 .0 0 2 p = 0 .0 0 1

p = 0 .0 0 2 p = 0 .0 0 1

PTCs have a low HIV reservoir distributed in the resting memory CD4 T cell subsets

Saez-Cirion et al Plos Pathogens 2013

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

PTC

PHIPatients at PHI :ANRS OPTIPRIM trial at inclusion

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

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

Institut PasteurRégulation des Infections Rétrovirales

Asier Saez-CirionGianfranco PancinoDaniel Scott-Algara

Françoise Barré-SinoussiPierre Versmisse

Faculté de Médecine Paris SudINSERM U1012Cécile Goujard

Alain VenetOlivier LambotteIsabelle Girault

Jean-François Delfraissy

INSERM U1018Laurence Meyer

Faroudy Boufassa

CHU Necker Enfants MaladesUniversité Paris Descares

Christine RouziouxVéronique Avettand-Fenoel

Adeline MélardAntoine Chéret

CHR Orléans La SourceService Maladies Infectieuses

Laurent HocquelouxThierry Prazuck

CHU Hôtel-DieuUniversité Paris Descatres

Jean-Paul ViardJade Ghosn

Patients and clinicians who participate to the cohort studiesCHU Pitié-SalpetriereINSERM UMR-S 945

Brigitte AutranCharline Bacchus

Benjamin DescoursAssia Samri

Ioannis TheodorouJulien Guergnon

ANRS CO6 ANRS CO15 ANRS CO18 Essai ANRS 147 PRIMO ALT HIV controllers OPTIPRIM

Acknowledgements

All Funded by the