Can T cells control the HIV and HCV infections?

Can T cells control the HIV and HCV
infections?
,
Impacts on the HIV reservoirs and HCV clearance
Brigi%e'AUTRAN'
Lab.'Immunité'et'Infec9ons,'INSERM'U945'
Ins9tut'Fédéra9f'de'Recherches''ImmunitéFCancerFInfec9on'
Hôpital'Pi9éFSalpétrière'
Université'Pierre'et'Marie'Curie,'Paris
'
brigi%[email protected]'
'
GISHEAL'
Can T cells control
the HIV and HCV infections?,
•  Impact on the HIV reservoirs in the ALT/ Elite Controllers:
–  Positive Impact on HIV reservoirs of:
–  Immune response genes
–  HIV-specific CD8 T cells
–  Positive Impact of HIV-specific CD8 T cells on the
distribution of the HIV reservoirs among CD4 T cell sub-populations
•  Impact on the HCV clearance during acute HCV hepatitis
in HIV co-infected patients:
–  Positive Impact of the HCV-specific CD4 memory T cells
–  Negative Impact of the HCV-specific T cells producing TGF-beta
2
HIV RESERVOIRS at various stages of the HIV infection
3
Kine%cs(of(early(HIV(infec%on((and(Cons%tu%on(of(the(HIV(Reservoirs(
( ((
( ( (from%A%Haase,%2010%
% % '%
%%%
CCR7'
memory(
Effector(
memory(
Memory(
CD4(T(cells(
Naive((
CD4+cell(
CD45RA'
CCR7'
HIV(Reservoirs(
CD45RA'
CD45RA'
effector(
effector(
Effecto
r(
CD4(T(
cells'
memory(
Transi%onnal(
memory(
CD4(T(cells(
CCR7'
Central(
memory(
memory(
Memory(
CD4(T(cells(
CCR7'
( ((
CCR7(
(((
CD27'
Apoptosis((
((
Turnover'
CD45RA'
Réponses(Immunes(
4
Positive impact of ImmuneGenetics (HLA) on the HIV Reservoirs
Genome-wide analysis in Elite Controllers and LTNP
HLA-B57 (HCP5)
!  HLA-B57 (HCP5)
is the strongest Marker
!  The HLA locus in Chromosome 6
for low HIV reservoirs
is the strongest genetic marker for:
•  HIV controllers vs Acute infection,(Dalmasso et al.:
•  European
(Dalmasso et al. 2009)
PLosOne, 2009)
LTNP (GISHEAL: ALT+ Italian cohort) vs Acute infection (Guergnon et al: J Inf. Dis. 2011))
5
'
ALT-LTNP ANRS CO-15 Cohort:
Distinct influences of various HLAs on HIV reservoirs
5
HIV-RNA
Copies/mL
HIV-DNA
Copies/Million PBMC
4
Median log10 HIV
P=0.01
3
P=0.003
2
1
0
+- +-
+- +-
+- +-
+- +-
6
ALT ANRS CO-15 cohort:
Impact of HIV-specific CD8 T cells on the HIV Reservoirs:
Distinct modalities of influence of HLA-B27- or B-57 on Gag-specific CD8 T cells
Descours &Avettand-Fenoel et al.
CID 2012
HLA-B27/57 restricted CD8 cell anti-HIV-Gag
GagHspecific(CD8+(T(cells(
Magnitude
B27 donors
GagHspecific(CD27+TCM(CD8+(T(cells(
HIV Reservoirs
and
HLA B27/57
Differentiation
B57+ donors
Functional
avidity
/Gag(epitopes(
Mar0nez%%V%et%al.%J.I.D.%2005;%Almeida%et%al.%J%Exp%Med%2007,%Blood%2009%
X%Jie%et%al.%AIDS%2010%
7
Study of the HLA-B27/57 Reservoirs
ALT Cohort: An ANRs cohort of LTNP
(71 individuals defined on CD4 counts in 1996,
followed for > 15 years)
•  Selection of 18 patients:
• 8 HLA-B27/57 LTNP, including 4 Elite
Controllers (LTNP-ECs):
• 10 non B27/57 including 4 LTNP-ECs:
•  Sorting of Resting naïve, TCM, TTM TEM and
Eff. CD4 T cells
(CD25-69-DR-; CD3+4+45RA+CCR7+CD27+)
•  Quantification of cell HIV-DNA in sorted subsets
!  Cell sorting of resting
(CD25-CD69-HLADR-) CD4 T cells
and HIV DNA quantification
Impact of HLA-B27 et B-57 on the immune distribution of HIV Reservoirs
in LTNPs:
Protection of Lymphocytes Central-Memory CD4 T cells
!  LTNP(HIV(Reservoirs(
! (HLAHB27+/57+:(Protec%on(of(CD4((TCM(
(TTM(>(TCM)(≥((TEM(>(TN)(
HLAHB27/57+(
HLA-B27/B57
Non(HLAHB27/57(
non HLA-B27/B57
0.0313
Measured cell HIV-DNA
Calculated threshold
TEM
p=0.0081
TTM
270'
TCM
120'
TN
195'
TEM
1319'
101
TTM
741'
102
TN
92'
103
TCM
cell HIV-DNA (copies
/millions cells)
104
0.0078
0.0078
0.0078
0.0469
9
Intensité(des(réponses(CD8(an%HHIV(est(associée(au(Contrôle(des(Reservoirs(VIH(dans(
les(Ly(T(CD4(Memoire(Centrale(
(
Descours;%AveIandJFenoel,%CID,%2012%
anti-HIV CD8 T Lymphocytes : key for the control of HIV Reservoirs inTCM
!  CD8 anti-HIV-Gag /Target cell
(E/T) ratio
in HLA-B27/57+ > HLA-B27/57-
E/T ratio In vivo :
= Nb CD8 anti-HIV / Nb cellules HIV DNA+
(si 1 copie HIV DNA /1 cellule infectée
! (Distribu%on(of(HIV(Reservoirs(Correlate(with(the((
CD8(an%HGag(/(Infected(CD4((E/T)(ra%o(
anti-HIV CD8 T Lymphocytes : key for the control of HIV Reservoirs inTCM
HLA-B27/57 neg
!  Critical protection of long-lived
TN
Antigens
TCM
Antigens
TTM
Antigens
TEM
Central- Memory CD4
HLA-B27/57pos
T Lymphocytes (TCM):
!  required for maintaining
TCM
TN
TEM
TTM
T cell Numbers and Function
Naive
CD4+cell
effector
effector
Effector
CD4 T
cells'
CCR7
CCR7
memory
Effector
memory
Memory
CD4 T cells
Apoptosis
in the long term
CD45RA
!  preserved in LTNP / Controllers
CCR7
CD45RA
memory
memory
Transitionnal
CD4 T cells
CCR7
Central
memory
memory
Memory
CD4 T cells
CCR7
CD45RA
Turnover'
CD27
CD45RA
HIV Transcriptionally active
HIV Transcriptionally inactive
B Descours et al CID 2012.
Contrôle Immun des Reservoirs HIV ?
Univ(Pierre(et(Marie(Curie(,'(Pi%éHSalpétrière,''
IFR(ImmunitéHCancerHInfec%on(
CRIV((Centre(de(Recherches(Intégrées(sur(le(VIH)(
'
Immunology,(INSERM(U945'''
Immunity'to'viruses
''''immunogene%cs((((((((HIV(immunosenescence((((((
A(Samri( ((((B(Descours ((((I(Theodorou( (
A(Guihot ((((V(Mar%nez(( (((J(Gueurgnon '
G(Carcelain
''
'
'
((V(Appay,(((
'J'Almeida ''
'
INSERM(U943( (Sta%s%cs(:''''D.'Costagliola'et'al.(
'
'
'Clinics:' '(((((C(Katlama,'R'Tubiana,'MA'Valan9n….'
'
'
'Virology:((((((V'Calvez,'AG'Marcellin'
ALT((ANRS(CO15(Cohort(and(Reservoir(study(group:''
Virology'and'serology':(C(Rouzioux,((V(Aveeand','Univ.'ParisF5'
'
Clinics:'
'JP'Clauvel,'SaintFLouis'Hosp.'Univ'ParisF7'
'D'Sicard,'Cochin'Hosp.'Univ.'ParisF5''''
12
Co-infection VHC/VIH
VIH
VHC
•  Frequent : 40% ; same risk factors
•  HIV Infection VIH untreated
•  Increase HCV viremia.
•  Modifies serology.
(Zylberberg Gastroenterol Clin Biol 1999)
•  Accelerates the HCV natural history:
•  Augments evolution towards chronicity: 85 %.
•  Aggravates hepatic lesions :
•  Progression faster towards cirrhosis 15- 22% (2,6 à 11%).
•  Decreases frequency of spontaneous recovery : 5 - 10%.
(Allory Y. et al, Hum Pathol 2000), (Soto,B. J Hepatol 1998), (Sulkowski M. et al, Ann Int Med 2003).
13
Can Immune responses to HCV control the virus ?
" HCV(MonoHinfec%on(:(
Immune'Responses'associated'with'spontaneous'clearance:'
!  CD4'and'CD8'T'cell'Responses''strong'and'diversified'
!  Cri9cal'role'of''CD4'Th1'cells''producing'IFNFγ,'ILF2,'TNFFα:''
•  Strong'prolifera9ve'responses'against'non'structural'proteins:''
•  Key'for'the'genera9on'of'an9FHCV:''CTL''and'neutralizing'Abs''
''
'
'
'Thimme%R%et%al,%J%Exp%Med%2001,%Cox%A%et%al,%Hepatology%2005,%Lechner%F%et%al,%J%Exp%Med%2000,%%
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%SmykJpearson%et%al,%J%Virology%2008,%Kaplan%et%al%Gastroenterology%2007%
((
" HIV/HCV(CoHinfec%on(:(
!  Weaker'immune'Responses %(danta%et%al,%JID,%2008)!
!  Memory'prolifera9ve'Responses'associated'with'spontaneous'clearance'
%%
%
%
%
%
%
%(Kim%AY%et%al.%PLoS%Med%2006)!
!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!
14
Etude ANRS HC EP 21 « C-gue »:
Characteris%cs(of(an%HHCV(T(cells(associated(with(recovery
!  Prospec%ve(study(over(15(months(of(HIV/HCV(coHinfected(Pa9ents''
fin de suivi
Absence de PCR
dans les 6 mois
avant diagnostic
RVR
fin de traitement
début de traitement
RVS
NR
PCR +
diagnostic
Hépatite C chronique
PCR +
Évolution sous traitement
Guérison sous traitement
PCR -
Évolution spontanée
Guérison spontanée
PCR -
0
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
Mois
Analyses immunologiques
15
Les cibles antigéniques des cellules T anti-HCV
Analyse par:
- test de prolifération : cellules T centrales mémoire
- Elispot-IFN-g : cellules T mémoires et effectrices
- Cytolines intra-cytoplasmiques ; IFN-g, IL-2, TNF-a, TGF-b, IL-10
16
Correlation between anti-HCV memory proliferative
T cell responses and recovery
A. Schnuriger et al. AIDS 2009
HCV load
6,2
6
5
4
3
2
1
0
6
5,8
1st Study:
N=38 patients
Spontaneous recovery (N)
5,6
5,4
5,2
Resp /
HCV
Non Resp
/ HCV
Resp /
HCV
Non
Resp /
HCV
Resp /
NS4
Non
Resp /
NS4
#  Prolifera%ve('Responses((centralHmemory(type)''
• 'Weak'in'frequency'and'intensity:'
• 'But'associated(with(spontaneous(clearance:((against(all(HCV(proteins(
(
(
(
(
(
(
(
(
(
(
(
(
(
(
(
((p=0,02)'
(((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((( (
(
(( ((((((((against(NS4((p=0,03)'
#  (Th1'(IFNFγ)'Responses(:(Not(correlated(with(clearance'
• 'weak'and'mostly'directed'against'NS3,'NS5'
• 'amplifiables'ajer'in'vitro'culture'(mostly(CD4+'(ILF2,'IFNFγ)
17
TGF-β, Immune responses and HCV
!  Produit'par:'cellules'T'CD4'et'CD8,'T'reg,'''
''''' '
'
''''''monocytes,'cellules'de'kupffer''
!  Régulateur(majeur(des(fonc%ons(immunitaires((
! ''Inhibe:''
•  Réponses'Th1,''
•  Fonc9ons'des'CTL'et'cellules'NK''
'''''des'cellules'dendri9ques'(DC)'
! 'Impliqué'dans'la'généra9on'des'cellules'T'reg'
! 'Induit'la'généra9on'des'cellules'Th17'en'présence'de'l'ILF6'
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%(Bhagelu%Ram%Achyut%et%al%%Gastroenterology%2011)%%
%
!  Rôle(proHfibro%que'(impliqué'dans'la'fibrose'hépa9que)'
!  Inhibe(la(réponse(an%HVHC(chez'les'pa9ents'VHC+'en'phase'chronique'(Rowan,%AG%J%
Immunol%2008,%Alatrakchi%N%et%al%J%%Virol%2007)%
18
Inhibition of anti-HCV Th1 responses during HIV co-infection ?
C-Gue Study 2
((
((
((
((
Total((n=24)(
CD4/mm3(médian((IQR)(((((((((((((((((((((((((((((((((((((((((((554((470H643)(
VIH(Viral(Load(VL(médian((IQR)((((((((((((((((((((((((((((((2,6((2,6H4,6)(
HAART(No((%)((((((((((((((((((((((((((((((((((((((((((((((((((((((((((19((83%)(((((((((((((((((((((((((((((((((((((
Infec%on(VHC(asymptoma%que(No((%)(((((((((((((((((((8((35%)((((((((
Peak(ALT(UI/mL(médian((IQR)(((((((((((((((((((((((((((((((((422((187H542)(
Log(VHC(VL(médian((IQR)(((((((((((((((((((((((((((((((((((((((6,1((5,3H6,8)(
Génotype(No((%)(
((
((
((
((
génotype(1H3((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((6((22%)(
((
génotype(4(((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((17((74%)(
((
NA((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((((1(
((
Groupe A (16)
Guéris (G)
5 spontaneous
recovery
11 responses
to therapy
Groupe B (8)
Chronique (CH)
4 patients chronic
hepatitis
4 non responders
to therapy
19
Th1 responses against HCV are inhibited by TGF-beta and IL-10
!  Weakness( of( IFNHγ( producing( Th1( cells( an%Hcore,( NS2,( NS4,( E1( lead( to( inves%gate(
the(inhibitory(role(of((TGFHβ(and/or(ILH10(of(T(cells(directed(against(Core,(NS2,(NS4,(E1(
IFN-γ SFC/106 PBMC
ELISpot!IFN+γ!with%Neutralizing!Abs!against!IL+10!and!TGF+β!!a?er!S0mula0on%with%HCV%pep0des%%%
150
250
E1
200
NS2
200
100
1
Isotypes
Isotypes mAbs
100
1
1
mAbs
200
0
0
2
300
50
50
0
400
100
100
P24
500
150
150
50
600
NS4
2
0
1
2
Isotypes mAbs
Isotypes mAbs
2
Isotypes
mAbs
Répondeurs:
0/10
2/10
1/10
3/10
0/9
2/9
2/10
2/10
4/8
5/8
!  (Blocking(ILH10(and(TGFHβ(:(
!  allows(produc%on'of'IFNHγ'against'HCV'in'4/11'pa9ents'tested,''
' '
'
'
''
'
'
'
'
'''''3/4'progressing'towards''chronic(infec%on'(
'
S Harfouch
et al. J Hepatol. 2012
20
TGFHβ(producing(an%HHCV(T(Cells(are(significantly(increased(in(pa%ents(progressing(
towards(chronicity(than(in(spontaneous(or(therapeu%c(recovery(
! 'TGFHβ'(but'not'ILF10)'producing(T(CD4+(and(CD8+'cells'against'HCV'Core'and'NS2:'
! 'are'detectable'in'8/24'tested'pa9ents'(33%)':'Median:''CD4+TGFFβ+'0.58(%'(IQR:'0.2F0.81)'
0.
1
HCV
core/NS2
'
medium
0
.
7
9
3
0
.
8
'
'
'
HCV
'
0
.
9
8
0
.
8
3
'''''''''CD8+TGFFβ+'0.64(%'(IQR:'0.27F0.78)'
p=0.03
p=0.04
1.0
0
.
2
0.
1
'
0.6
0.4
0.2
0.6
0.4
0.2
0.0
HC: 8
R:16
Healthy Recovery
S Harfouch et al. J Hepatol. 2012
p=0.01
0.8
0.8
0.0
TGF-β
p=0.04
1.0
%CD8+ TGFβ+
CD4
0
.
2
'
CD8
medium
'
%CD4+ TGFβ+
'''''''''''''''''''''''''' '
Controls
Ch: 8
Chronic
HC:8
R:16
Healthy Recovery
Ch:8
Chroni
Controls
21
Lack of TGF-β producing anti-HCV T cells (core/NS2) are associated with
N patients
N patients
spontaneous or therapeutic recovery, independently of the IL-28B polymorphism
TGF-β+
TGF-β-
TGF-β+ T Cells anti-HCV
S Harfouch et al. J Hepatol. 2012
C/C
C/T+T/T
IL-28B Polymorphism
22
Conclusion: T cells specific for HCV associated with
recovery during acute HCV hepatitis
in HIV co-infection
1H(Presence(of((centralHmemory)(prolifera%ng(CD4(T(cells(an%HHCV((NS4)
! 'is'associated'with'spontaneous(HCV(clearance(
!
!
!
!
!
!
!
!
!
!
!
!
!
!A!Schnuriger!AIDS!2009!
2H(Lack(of(TGFHβ(producing(T(cells(an%HHCV(core/NS2('predict(
spontaneous(or(therapeu%c(HCV(clearance((independently(from(
ILH28B((C/C)'polymorphism(and(plasma'levels'of'TGFHβ'
S Harfouch et al. J Hepatol. 2012
23
Can T cells control
the HIV and HCV infections?,
•  HIV : In the ALT/ Elite Controllers: Yes but only a
–  Positive Impact of
• 
Immune response genes on HIV reservoirs
•  HIV-specific CD8 T cells on HIV reservoirs
–  Positive Impact of HIV-specific CD8 T cells on the
distribution of the HIV reservoirs among CD4 T cell sub-populations
•  HCV: during acute HCV hepatitis in HIV co-infected patients:
– 
Impact on the HCV clearance :
•  Positive for HCV-specific CD4 memory T cells
•  Negative for HCV-specific T cells producing TGF-beta
•  The Janus Head: Why the same cells have different effects ???
24
The C-Gue Studies
Département(d’Immunologie(Cellulaire((
Pi%éHSalpêtrière(URMS(945:'
B.'Autran,''
A.'Samri'
I.'Theodorou'
A.'Schnuriger'
S'Harfouch'
'
Laboratoire(de(Virologie(Pi%éHSalpêtrière(:'
V.'Thibault'
'
'
'
Unité(de(sta%s%ques(Pi%éHSalpêtrière(:'
M.'Guiguet'
Service(de(Maladies(Infec%euses(Pi%éHSalpêtrière(:(
S.'Dominguez,'MA.'Valan9n'et'R.'Tubiana'
'
'
'
Service(de(Médecine(Interne(Pi%éHSalpêtrière(:'
A.'Simon'
'
'
'
Service(de(Maladies(Infec%euses(St(Antoine(:'
PM'Girard,'Z.'Ouazène'et'P.'Campa.'
'
'
'
Service(de(Maladies(Infec%euses(Tenon(:'
L.'Slama'et'P.'Bonnard.''
'
'
25