Towards a therapeutic vaccine to reduce HIV reservoir, both neutralizing HIV and

inhibiting effector T cells depletionPr. Patrice DEBRE

Laboratoire Immunité & InfectionLaboratoire Immunité & InfectionINSERM UMR-S 945INSERM UMR-S 945

Hôpital Pitié-SalpêtrièreHôpital Pitié-SalpêtrièreParis, FranceParis, France

“Towards an HIV Cure” Pre-Conference Symposium20 & 21 July 2012

A strategy to decrease HIV reservoir

INFECTION

Virus Infected CD4 cells

PATHOGENESIS

NK Non-infected

CD4 cells

Vaccine

VirusNeutralizatio

n

Restore Immune-response

An epitope based vaccine, both

1) Inhibiting the deleterious effect of NK cells

A NK ligand on CD4+ T cells ?

2) Neutralizing HIV-1

NKp44L is induced during HIV infection by the 3S/gp41 motif

Kill

NK

CD4

+-NKp44

MHC-I NKp44L

%

NK

p4

4L

010

20

3040

3- 3-4+ 4+8+ 8+

Control PBMCHIV+ PBMC

CD4 / mm3

0 250 500 750 1000010

20

30

40

p=0.002

%

NK

p4

4L

HR1 HR2Fusion Peptide

TM517 532 558 595 643 678

3S motif3S motif

HIV-1 gp41 Env

3S : very conserved motif & specific to HIV-1

NKp44L

2.7% 17.4%

3SUT

CD

4

Vieillard et al Proc Natl Acad Sci USA 2005

gC1qR is the cell-surface receptor of the 3S/gp41 motifon CD4+ T cells

HK

3S

C1q

C3

NA0 25 50 75 100 125 150 175

NKp44L expression (MFI) NKp44L expression

IgMNA

3S

3S+ agC1qR

Role of gC1qR !

Listeria monocytogenesStaphylococcus aureusPlasmodium falciparum

HCV, rubella virus, HSV and HTNV

Fausther-Bovendo et al PLoS Pathogens 2010

Deleterious effect of NK

cells ?

Anti-3S Ab : a predictive value for the evolution of the disease ?

0

100

200

300

0 400 800 1200

Anti-3S (U/mL)

CD4 count/mm3

Vieillard et al AIDS (2006)Unpublished data

Multivariate study between 40 patients (first quartile) with a slope CD4>2.7/month and 40 patients (last quartile) with slope CD4<-6.9/month

Variable Odds-ratio IC95 P

Sexe M WAge (10y)

CD4 (X100)

Log VL

3S Ab (x100)

1.00 1.83 1.30

0.58

1.17

2.89

0.049-6.87

0.76-2.23

0.42-0.81

0.55-2.51

1.60-5.17

0.372

0.334

0.001

0.682

<0.001

Summary-1Summary-1

- NKp44L is specifically expressed on non infected CD4+ T cells from HIV-1 patients;

- NKp44L is induced by a specific peptide from the gp41 HIV-1 protein;

- Anti-3S antibodies are predictive of the disease evolution.

A 3S vaccine inhibiting HIV pathogenesis

1- Macaque model of SHIV infection1- Macaque model of SHIV infection(Vieillard et al, AIDS 2008)(Vieillard et al, AIDS 2008)

2- Prophylactic vaccination2- Prophylactic vaccination(Vieillard et al, PNAS 2008)(Vieillard et al, PNAS 2008)

3- Therapeutic vaccination3- Therapeutic vaccination(Vieillard et al, submitted)(Vieillard et al, submitted)

Proof of concept in macaques

Collaboration: Roger Le Grand (CEA, Fontenay-aux-roses; France)

Prophylactic vaccination by the 3S-gp41 peptideProphylactic vaccination by the 3S-gp41 peptide

Vieillard et al Proc Natl Acad Sci USA (2008)

D385

InfectionSHIV162P3

D0

Immu 1

D30

Imm. 2

D60

Imm. 3

D585

Animal groups :

Analysis:

• Anti-3S antibodies • Plasma viral load• Blood lymphocyte phenotyping• NK & CD4 activation• Cytotoxicity• Apoptose• Inflammation

Sacrifice

Study design

KLH-control macaques :

3S-immunized macaques :

Effect of 3S vaccination on Viral load, NKp44L expression, Effect of 3S vaccination on Viral load, NKp44L expression, and CD4 countand CD4 count

CD

4 /

mm

30

500

1000

1500

2000

0 50 100 150 200

** *

Vir

al lo

ad

1

102

104

106

108

0 50 100 150 200

NS NS

0

1020

30

40

50

0 50 100 150 200

% C

D4

+N

Kp

44

L+

0

500

1000

1500

-400 -200 0 200

an

ti-3

S (

U/m

L)

3S

3S3S SHIV162P3

Days post-infection

KLH-control macaques 3S-immunized macaques

3S

3S

3S

3S

Effect of 3S vaccination on CD4 activation & InflammationEffect of 3S vaccination on CD4 activation & Inflammation

0 20 40 60 80Days post-infection

% C

D4

+C

D6

9+

0

2.5

5

7.5

10

KLH-control macaques 3S-immunized macaques

Days post-infection70

1020304050

CR

P (

g/ml)

**

*

0 210

20

40

60

80

0

TN

F-

(pg/ml)

**

7 21

Summary-2Summary-2

• Immunogenicity in cynomolgus (anti 3S response)

• BUT Unchanged viral load

• Decreased NKp44L expression on CD4+ T cells• Decreased NK cells cytotoxicity on CD4+ T cells• Decreased CD4+ T cells apoptosis• Preservation of CD4+ T cell counts• Decreased immune activation• Decreased inflammation during the acute phase

Summary

Epitope 3S(SWSNKS)

CD4+ T cell

HIV infected cells

HIV virion

gp41

NKp44L

CD4+ T cell lysis

Autologous NK cells

Anti-3S Ab

NKp44

CD4+NKp44L+ cell

E/T ratio

0

10

20

30

40

100/150/125/112.5/1

44L+

44L-

% N

K

Lysis

CD4 depletion

Cell activation

Inflammation

An epitope based vaccine, both

1) Inhibiting the deleterious effect of NK cells

2) Neutralizing HIV-1

Search for an effect of 3S mutant

NH2-SWSNKS-NH2

Alanine scanning

HIV infectivity of 3S mutants

Infectivity

0

25

50

75

100

WT S613A W614A S615A N616A K617A S618A

% p24 production

Entry

%-galactosidase

activity

0

25

50

75

100

WT S613A W614A S615A N616A K617A S618A

Neutralizing effect of anti-3S mutant Abs

200

100

50

150

00 5 10 15 20

Ig (g/ml)

200

100

50

150

0

Human AbsMice Abs

200

100

50

150

00 5 10 15 20

Ig (g/ml)

p24 (ng/ml)

00 4 6 8 10

Days post-infection

2

200

100

50

150

p24 (ng/ml)

WA

WA

WT

WT

WT

WT

W/o Ab

#109

#109

p24 (ng/ml)

0 4 6 8 10

Days post-infection

2

Antibodies against 3S (WA) mutants also inhibit NK pathogenesis

NT 3S WT

#117 #24 #44

#65 #71 #109

NKp44L

68.3 6.4

21.9 34.6 28.4

21.9 31.3 22.7

CD107a

NT 3S WT

38.3 58.2

2.90.6

29.4 34.1

28.67.9

32.1 64.2

2.11.6

#117 #24 #44

30.3 55.1

1.81.8

40.9 52.1

4.92.1

29.9 61.0

7.71.4

#65 #71 #109

NKp44

30.4 62.6

5.91.1

36.4 55.0

6.32.3

34.9 55.4

7.81.9

ConclusionConclusion

1) Restoring/preserving CD4 cell functions

A gp41 epitope based vaccine, both

2) Neutralizing HIV-1

As a tool to decrease HIV reservoir

In Progress …In Progress …

Animal Model Clinical trialPhase I/IIa

HIV-1 infection

Non-infectedAIDS

Asymptomatic phase

TherapeuticVaccine

Antiretroviral Therapy

Unité Immunité & Infection

Team-2

Patrice Debré

Hugues Fausther Bovendo

Caroline Petitdemange

Alexis Sennepin

Abla Achour

Florence Baychelier

Vincent Vieillard

Team-1

Brigitte Autran

Assia Samri

Unité Epidémiologie

CliniqueDominique Costagliola

Service de Virologie

Henri AgutVincent CalvezDaniel CandottiIsabelle Malet

Service des Maladies

Infectieuses & Tropicales

Christine Katlama

Hôpital Pitié-Salpêtrière - Paris

Institut Pasteur

Olivier Schwartz Nathalie Sol-Foulion

Felix Rey

CEA

Laboratoire d’ Immuno-VirologieRoger Le Grand

Nathalie Deudreude-Bosquet

Aurélien Corneau

Isabelle Mangeot-Méderlé

… Grants

ANRS

Sanofi-Pasteur

Bill & Melinda Gates Foundation

Agence Nationale de Recherche (ANR)

InnaVirVax

ORVACS

PartnersPartners