immunosénescence et vih€¦ · immunosénescence et vih victor appay infectious diseases and...
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Immunosénescence et VIH
Victor Appay
Infectious Diseases and Immunology Research Center (CIMI)
INSERM, Hôpital Pitié Salpêtrière, Paris , France
Fight Aging and STress department (FAST)
SORBONNE Université, Paris, France
International Research Center for Medical Sciences (IRCMS)
Kumamoto University, Kumamoto, Japan
2
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Intervenant :Victor Appay
Titre : Immunosenescence et VIH
Course of HIV infection
Viral load
Acute infection Chronic infection
Asymptomatic period AIDS and death
6 - 12 weeks 1 - 25+ years 2 - 3+ years
CD4+ T-cells
HIV specific CD8+ T-cells
Neutralizing antibodies
? ?
=> better understand disease progression…
Innate immunity
HIV infection and replication Main target: activated CCR5+ CD4+ T lymphocytes
Chronic immune activation (Adaptive and Innate Immunity)
Collapse of the immune system / AIDS
Immune activation and progression towards AIDS
Causes Massive depletion of CD4+ T cells particularly in gut-associated lymphoid tissue
Bacterial
translocation TLR ligands
Viral
reactivation in particular CMV
Production of
HIV proteins gp120, nef, tat
Immune response
against HIV T, B and NK cells
HIV infection and replication Main target: activated CCR5+ CD4+ T lymphocytes
Chronic immune activation (Adaptive and Innate Immunity)
Collapse of the immune system / AIDS
Immune activation and progression towards AIDS
Consequences ? ?
Immune exhaustion /
Immunosenescence?
Naïve Antigen experienced
IL-2 IFN-
Early Late Intermediate
CD27+
CD28+
CCR7+
CD45RA+
CD27+
CD28+
CCR7+
CD45RA-
CD27+
CD28+
CCR7-
CD27+/-
CD28-/+
CCR7-
CD27-
CD28-
CCR7-
Central memory
Effector memory
Subpopulations of T lymphocytes (CD4 & CD8)
Ph
en
oty
pe
Differentiation
Naive
+Antigen +Ag -Antigen -Ag
“Effector” “Memory” “Effector” “Memory”
Pri
ma
ry
Infe
cti
on
Se
co
nd
ary
Infe
cti
on
Identification
by flow cytometry
CCR7+
CD28+
CD27+
CD57-
CCR7-
CD28+
CD27+
CD57-
CCR7-
CD28-
CD27+
CD57-
CCR7-
CD28-
CD27-
CD57+
Telomere lenght
Proliferative capacity
TCR repertoire diversity
CDR3
length
CFSE fluo Telomere
MFI
Earl
y
La
te
Inte
rmed
iate
C
entr
al
mem
ory
E
ffecto
r m
em
ory
Mem
ory
T c
ell
dif
fere
nti
ati
on
Memory T cell subsets & Aging features
1. Initial evidence of immune aging in HIV infected patients:
High frequency of highly differentiated / old memory T cells
Effros RB et al, AIDS, 1996
Brenchley et al, Blood, 2003
Papagno et al, Plos Biol, 2004
with age and HIV-1 infection
=> CD28- CD57+ T cells => “marker of immunosenesence”
Is increased frequency of CD28-/CD57+ T cells such a relevant marker of immune aging and HIV disease progression?
=> No clear association with disease progression
Absolute numbers Evolution with ART
Longitudinal Transversal
What drives the accumulation of CD57+ memory T-cells?
=> CMV specific CD8+ T cells
are highly differentiated
=> In HIV+ patients (specially ART+):
strong CMV specific responses
=> Memory inflation C
MV
in
fecti
on
Gated on tetramer+
CD8+ T cells
CMV => Confounding factor of the immune parallel
between Aging and HIV disease progression
-Impact on immune profile in HIV infected patients
=> Main driver of accumulation of CD28-/CD57+ T cells
CMV co-infection in HIV infected patients
CMV => Risk co-factor of multiple comorbidities in HIV infected patients
- Impact on mortality (non AIDS events)
=> Infants born from CMV/HIV coinfected women
=> CMV/HIV coinfected adult patients (Slyker, AIDS 2009; Lichtner, JID 2015)
- Impact on chronic immune activation and inflammation
(increased CD38+ T cells, IL-1b, IL-6, IP-10 levels) (Hunt, JID 2011, Freeman, CID 2016)
=> Influence of immune reconstitution capacity (Appay, AIDS 2011)
=> Co-infections e.g. HCV (Kuniholm, Plos One 2013)
=> Comorbidities: cardiovascular and neurocognitive disorders
Carotid artery intima-media thickness & CMV IgG or T cells (Hsue, AIDS 2006; Parrinello, JID 2012)
Premature immune aging in HIV infection
Old memory T cells
Healthy donor
HIV infected
Non progressor
Healthy elderly
HIV infected
Treated
Accumulation of
end stage memory
lymphocytes (CD28- / CD57+
Short Telomere,
Reduced proliferation)
Naive T cells
?
?
?
HIV infected
Progressor
2. Collapse of naive T cell frequencies
with HIV disease progression and aging
Absolute numbers Evolution with ART
Longitudinal Transversal
=> Strong association with HIV disease progression
and immune aging
CD8+ T cells (effector functions: IFN-g+TNF
Perforin+Granzyme)
Control of HIV
replication and reservoir
Impact of immunosenescence on capacity to mount
new CD8+ T cell responses in HIV infected patients?
Vaccines
and
Immunotherapies
Clonal loss
& exhaustion
=> Key role of CD8+ T cell naïve pool and priming capacity to fight HIV
Genomic instability of HIV: Creation of neoantigens
Escape of immune recognition
CD8+ T-cell priming efficacy and HIV infection?
- Simple human setting
- Using little biological materials
- Large panel of donors (HLA-A2) => Large comparative studies of human samples
III. Advantages
Healthy HLA-A2 blood donor
High frequency of Melan-A specific naïve CD8+ T cells
• ~2.10-4 circulating Melan-A specific CD8+ T cells
• CCR7+ CD45RAhigh CD45RO- CD28+ CD27+
• High levels of TRECS
• Long telomeres
• Polyclonal
Me
lan
-A
HIV
Ga
g
HIV
Po
l
CM
V p
p6
5
HC
V N
S3
An
tig
en
sp
ecif
ic n
aiv
e
CD
8+
T c
ell f
req
uen
ecy
I. Founding element
Melan-A / ELA = antigen model
Measure the efficacy to induce a de novo CD8+ T cell response
=> use of a simple in vitro model of naïve CD8+ T cell priming
Analysis of attributes
of primed Melan-A specific CD8+
T cells:
- Expansion capacity
- Phenotype
- Functional attributes
- TCR avidity
II. Experimental principle
FLT3L
=> Quantitative reduction
of CD8+ T cell priming
in HIV infected patients
Frequency of in vitro expanded tetramer+ cells => Priming efficacy
Assessment of in vitro CD8+ T cell priming efficacy in HIV-1-infected donors (with low viral replication)
Qualitative analysis of primed CD8+ T cells
FLT3L
+ Cytokines
FLT3L
+ HIV ssRNA40
(= TLR8L)
HIV+ prog
PBMC
Tb
et
Gra
nz B
Pe
rf
% M
elA
+ C
D8
T c
ells
HIV-
PBMC
* ** ** *
=> Qualitative reduction of T cell priming efficacy
(similar to T cell priming with old donor PBMC)
Priming conditions:
Premature immune aging in HIV infection
Naive T cells
Old memory T cells
Healthy donor
HIV infected
Non progressor
Thymus
Healthy elderly
HIV infected
Treated
Hematopoietic Progenitors / Stem cells
Accumulation of
end stage memory
lymphocytes (CD28- / CD57+
Short Telomere,
Reduced proliferation)
HIV disease
progression
=> Exhaustion of
primary immune
ressources (naïve cells)
?
?
?
HIV infected
Progressor
Immunological failure with ART
Aim => Characterize the circulating HPC compartment in HIV-1 infected
patients with good or poor immune reconstitution under ART
Co
ntr
ol
Lo
w C
D4
Lo
w C
D4
Hig
h C
D4
0
20
40
60
80
100
Lo
w C
D4
Lo
w C
D4
Hig
h C
D4
101
102
103
104
105
106
107
Lo
w C
D4
Lo
w C
D4
Hig
h C
D4
0
200
400
600
800
1000
Co
ntr
ol
Lo
w C
D4
Lo
w C
D4
Hig
h C
D4
0
200
400
600
800
1000
1200
1400
1600
HIV- HIV+
Ctrl No Tx NIR IR
HIV+
No Tx NIR IR
HIV- HIV+
Ctrl No Tx NIR IR
HIV+
No Tx NIR IR
Age
(yea
rs)
Vir
al lo
ad
(co
pie
s/m
l)
CD
4 n
adir
(cells
/ul)
CD
4 c
ount
(cells
/ul)
IR: Immune Responder
(good immune reconstitution
under ART)
NIR: Non Immune Responder
(poor immune reconstitution
under ART)
Disruption of the hematopoietic compartment in treated HIV-1 infected patients
lymphoi
d
myeloi
d
CD45RA
**
CLPcount(cell/µl)
Y M O0.01
0.1
1
10
**
HIV-
Age
HIV+
CD4
IRNIR Y M O0.01
0.1
1
10*
RatioLymphoid/M
yeloid
HIV-
Age
*
HIV+
IRNIR
CD4
B
=> NIR = low CD34 cell frequency, in particular CLP (lymphopoiesis)
CD34+ cell counts
NIR IR
Phenotype (lymphoid vs myeloid)
CLP
CMP
NIR
IR
HIV- HIV+
Mid Old NoTx NIR IR
CD
34 c
ount
(cells
/ul)
* **
CLP
count
(cells
/ul)
HIV- HIV+
Mid Old NoTx NIR IR
* **
Clonogenic potential of CD34+ hematopoietic progenitors
=> Lymphopoietic and reconstitution capacity of donors
+IL-7 +Flt-3L +SCF +TPO
FACS
analysis of
phenotype
Coculture for 4 weeks
Day 7, 14, 21, 28
Day 0
CD45RA
CD
7
CD34+
TLP
=> Reduced and skewed lymphopoietic capacity and increased
mortality (pyroptosis) of hematopoietic progenitors from NIR
Impaired lymphopoietic capacity of CD34+ cells from treated HIV-1 infected patients
Altered hematopoiesis and inflammation
P < 0.01
r = -0.42
% C
D3
8+
in
CD
8+
me
m T
ce
lls
CD34+ cell count (cell/µl)
0 1 2 3 4 50
20
40
60
80
Pla
sm
a s
CD
14
(p
g/m
l)
CD34+ cell count (cell/µl)
0 1 2 3 4 50
1000
2000
3000
4000
P < 0.0001
r = -0.51P < 0.01
r = -0.42
% C
D3
8+
in
CD
8+
me
m T
ce
lls
CD34+ cell count (cell/µl)
0 1 2 3 4 50
20
40
60
80
Pla
sm
a s
CD
14
(p
g/m
l)
CD34+ cell count (cell/µl)
0 1 2 3 4 50
1000
2000
3000
4000
P < 0.0001
r = -0.51
14 28 14 28 14 28 14 280.0
0.5
1.0
1.5
2.0
2.5
3.0
Control IL-6 IP-10 LPS
* * ns ns
Exp
an
sio
n le
ve
l o
f C
LP
(/ D
7)
Differentiation Altereddifferentiation
A
P < 0.01
r = -0.42
% C
D3
8+
in
CD
8+
me
m T
ce
lls
CD34+ cell count (cell/µl)
0 1 2 3 4 50
20
40
60
80
Pla
sm
a s
CD
14
(p
g/m
l)
CD34+ cell count (cell/µl)
0 1 2 3 4 50
1000
2000
3000
4000
P < 0.0001
r = -0.51P < 0.01
r = -0.42
% C
D3
8+
in
CD
8+
me
m T
ce
lls
CD34+ cell count (cell/µl)
0 1 2 3 4 50
20
40
60
80
Pla
sm
a s
CD
14
(p
g/m
l)
CD34+ cell count (cell/µl)
0 1 2 3 4 50
1000
2000
3000
4000
P < 0.0001
r = -0.51
14 28 14 28 14 28 14 280.0
0.5
1.0
1.5
2.0
2.5
3.0
Control IL-6 IP-10 LPS
* * ns ns
Exp
an
sio
n le
ve
l o
f C
LP
(/ D
7)
Differentiation Altereddifferentiation
A
Reduced in vitro TLP induction in the presence of inflammatory molecules
Ex vivo association between CD34+ cell numbers and inflammation markers
Inflammation => impaired lymphopoiesis = EXTRINSIC FACTOR
M O IR NIR0
5
10
15
20
25ns
***
**
***R
TL
CD
34
+ C
ell (K
b)
M O IR NIR0
1
2
3
4*
***
**
***
RT
A C
D3
4+
ce
lls (R
atio
)
C
Evidence of senescence in hematopoietic progenitors from HIV infected patients
M O IR NIR0
5
10
15
20
25ns
***
**
***
RT
L C
D3
4+
Ce
ll (K
b)
M O IR NIR0
1
2
3
4*
***
**
***
RT
A C
D3
4+
ce
lls (R
atio
)
CTelomere length
(qPCR)
Telomerase activity (TRAP assay)
Hematopoietic progenitors in NIR => approach senescence = INTRINSIC FACTOR
Co
ntr
ol
Lo
w C
D4
Lo
w C
D4
Hig
h C
D4
0
100
200
300
400
500
600
700
800
HIV- HIV+
Ctrl No Tx NIR IR
Co
un
t (c
ells
/ul)
Co
ntr
ol
Lo
w C
D4
Lo
w C
D4
Hig
h C
D4
0
100
200
300
400
500
600
HIV- HIV+
Ctrl No Tx NIR IR
Co
un
t (c
ells
/ul)
Impact on naïve T cell compartement in HIV infected patients
Naïve CD4+ T cells
Naïve CD8+ T cells
=> Legacy from HPC to naïve T cell compartment in NIR
Absolute counts Telomere length
Premature immune aging in HIV infection
Hematopoietic progenitors
Old memory T cells
Healthy donor
HIV infected
Non progressor
Bone marrow
Thymus
Healthy elderly
HIV infected
Treated
Accumulation of
end stage memory
lymphocytes (CD28- / CD57+
Short Telomere,
Reduced proliferation)
HIV disease
progression
=> Exhaustion of
primary immune
ressources (naïve cells &
hematopoietic
progenitors)
Naive T cells
HIV infected
Progressor
Causes Massive depletion of CD4+ T cells particularly in gut-associated lymphoid tissue
Bacterial
translocation TLR ligands
Viral
reactivation in particular CMV
Production of
HIV proteins gp120, nef, tat
Immune response
against HIV T, B and NK cells
HIV infection and replication Main target: activated CCR5+ CD4+ T lymphocytes
Chronic immune activation (Adaptive and Innate Immunity)
Collapse of the immune system / AIDS
Immune activation and progression towards AIDS
Consequences
Consequences
Causes Massive depletion of CD4+ T cells particularly in gut-associated lymphoid tissue
Bacterial
translocation TLR ligands
Viral
reactivation in particular CMV
Production of
HIV proteins gp120, nef, tat
Immune response
against HIV T, B and NK cells
HIV infection and replication Main target: activated CCR5+ CD4+ T lymphocytes
Chronic immune activation (Adaptive and Innate Immunity)
Collapse of the immune system / AIDS
Appay and Sauce, J Pathol, 2008
Secretion of
proinflammatory
cytokines
e.g. IL-6, TNF, IL-1, IP-10
Immunosenescence / Systemic aging
Inflammation
related disorders osteoporosis
atherosclerosis
neurocognitive deterioration
frailty
Inflam-aging
Sustained cell death
Turnover and Senescence
Homeostasic mechanisms
Immune cell renewal
Hematopoiesis
Exhaustion of
immune resources decline of regenerative capacity
loss of effective anti-HIV immunity
Immune activation and progression towards AIDS
Implications:
“Unhealthy vs. healthy” HIV infection
Viral replication &
Chronic immune
activation
Waning
immune
resources
Weakened immune
responses &
Imm reconstitution
VICIOUS CIRCLE
Low VL &
Limited immune
activation
Preserved
immune
resources
Potent immune
responses &
Imm reconstitution
VIRTUOUS CIRCLE
vs. Non progression (HIV-1 controller or
HIV-2 infection)
Disease progression (Classic HIV-1 Infection
+ co-infections)
=> Promote preservation of primary immune resources in HIV-1 infected patients:
- Early immune control (vaccine) - Early ART initiation
- Restore hematopoietic resources (pharma, reprogramming)
Collaborators Acknowledgements
Delphine Sauce
Laura Papagno
Mariela Piccin
Helene Vallet
Valérie Martinez
Alumni:
Solene Fastenackels
Kris Ragon
Véronique Fabre
Charles Bayard
Tinhinane Fali
Anna Lissina
Olivia Briceno
Francesco Nicoli
VA team INSERM Paris
INSERM Cochin, Paris
✓Roberto Mallone
✓Georgia Afonso
Institute of Infection & Immunity, UK
✓David Price
✓Emma Grant
Institut Pasteur, Paris
✓Asier Saez Cirion
✓Mathieu Angin
✓Jacques Boddaert
✓Judith Cohen Bittan
Geriatrics, Hôpital Pitié
Salpêtrière, Paris
Patients and
volunteers
Kumamoto University, Japan
✓Masafumi Takiguchi
✓Nozomi Kuze
Infectious diseases, Paris
✓ Christine Katlama
✓ Roland Tubiana
✓ Olivier Lambotte
✓ Jean Paul Viard
✓ Sophie Matheron
✓ Yasmine Dudoit
✓ Dominique Costagliola
Colliding cell cycle/mTOR, cell death, and senescence pathways
in hematopoietic progenitors from the elderly (HIV-)
Telomere shortening
+
Inflammatory factors (cytokines, PAMPs)
=> Cell cycle arrest /
Senescence
ROS
P2X7
Caspase 1 ATP => Pyroptosis
related programmed
cell death
NF-kB
(Fali et al, JCI insight, in press)
Impaired
lymphopoiesis
Cell cyling
Old
he
ma
top
oie
tic
pro
ge
nit
ors
Clonogenic potential of CD34+ hematopoietic progenitors
=> Lymphopoietic and reconstitution capacity of donors
+IL-7 +Flt-3L +SCF +TPO
FACS
analysis of
phenotype
Coculture for 4 weeks
Day 7, 14, 21, 28
Day 0
CD45RA
CD
7
CD34+
TLP
=> Reduced and skewed lymphopoietic capacity and increased
mortality (pyroptosis) of hematopoietic progenitors from NIR
Impaired lymphopoietic capacity of CD34+ cells from treated HIV-1 infected patients
CD1a
CD
5
PreT1 ProT2
ProT1
Pre-T
immature
7 14 21 28 7 14 21 28 7 14 21 280
5
10
15
20
25
30
HIV- HIV+
IR NIR
** ns
*
% o
f C
ells
M
7 14 21 28 7 14 21 28 7 14 21 280
20
40
60
80
100
120
ProT1
ProT2
PreTi
PreT1
HIV- HIV+
IR NIR
% o
f C
ells
M
CLPinduction CLPdistribution
7 14 21 28 7 14 21 280
20
40
60
80
100
120
PreTimmature
Old
B C
Differentiation
+
-
CLPdistributionatday14
31%
18%
35%
16%
14
ProT1
ProT2
PreTimmature
PreT1
8%
15%
28%
49%
14
ProT1
ProT2
PreTimmature
PreT1
17%
19%
35%
29%
14Y+IL-6
ProT1
ProT2
PreTimmature
PreT1
19%
15%
35%
31%
14Y+IP-10
ProT1
ProT2
PreTimmature
PreT1
18%
15%
40%
27%
14Y+TNF
ProT1
ProT2
PreTimmature
PreT1
Young Old
IL-6 IP-10 LPS
Young
Control
Day14
Differentiation
-
+
Middle-aged Old Middle-aged(HIV-)D28
8%
26%
29%
37%
7 14 21 28 7 14 21 280
20
40
60
80
100
120
ProT1
ProT2
7 14 21 28 7 14 21 280
20
40
60
80
100
120
ProT2
PreTimmature
7 14 21 28 7 14 21 280
20
40
60
80
100
120
PreTimmature
7 14 21 28 7 14 21 280
20
40
60
80
100
120
PreT1
Differentiation
+
-D28
7%
11%
21%61%
IR NIR
HIV+
B
CLPdistributionatday14
31%
18%
35%
16%
14
ProT1
ProT2
PreTimmature
PreT1
8%
15%
28%
49%
14
ProT1
ProT2
PreTimmature
PreT1
17%
19%
35%
29%
14Y+IL-6
ProT1
ProT2
PreTimmature
PreT1
19%
15%
35%
31%
14Y+IP-10
ProT1
ProT2
PreTimmature
PreT1
18%
15%
40%
27%
14Y+TNF
ProT1
ProT2
PreTimmature
PreT1
Young Old
IL-6 IP-10 LPS
Young
Control
Day14
Differentiation
-
+
Middle-aged Old Middle-aged(HIV-)D28
8%
26%
29%
37%
7 14 21 28 7 14 21 280
20
40
60
80
100
120
ProT1
ProT2
7 14 21 28 7 14 21 280
20
40
60
80
100
120
ProT2
PreTimmature
7 14 21 28 7 14 21 280
20
40
60
80
100
120
PreTimmature
7 14 21 28 7 14 21 280
20
40
60
80
100
120
PreT1
Differentiation
+
-D28
7%
11%
21%61%
IR NIR
HIV+
B
HIV
+
HIV
-
Altered hematopoiesis and inflammation
P < 0.01
r = -0.42
% C
D3
8+
in
CD
8+
me
m T
ce
lls
CD34+ cell count (cell/µl)
0 1 2 3 4 50
20
40
60
80
Pla
sm
a s
CD
14
(p
g/m
l)
CD34+ cell count (cell/µl)
0 1 2 3 4 50
1000
2000
3000
4000
P < 0.0001
r = -0.51P < 0.01
r = -0.42
% C
D3
8+
in
CD
8+
me
m T
ce
lls
CD34+ cell count (cell/µl)
0 1 2 3 4 50
20
40
60
80
Pla
sm
a s
CD
14
(p
g/m
l)
CD34+ cell count (cell/µl)
0 1 2 3 4 50
1000
2000
3000
4000
P < 0.0001
r = -0.51
14 28 14 28 14 28 14 280.0
0.5
1.0
1.5
2.0
2.5
3.0
Control IL-6 IP-10 LPS
* * ns ns
Exp
an
sio
n le
ve
l o
f C
LP
(/ D
7)
Differentiation Altereddifferentiation
A
CLPdistributionatday14
31%
18%
35%
16%
14
ProT1
ProT2
PreTimmature
PreT1
8%
15%
28%
49%
14
ProT1
ProT2
PreTimmature
PreT1
17%
19%
35%
29%
14Y+IL-6
ProT1
ProT2
PreTimmature
PreT1
19%
15%
35%
31%
14Y+IP-10
ProT1
ProT2
PreTimmature
PreT1
18%
15%
40%
27%
14Y+TNF
ProT1
ProT2
PreTimmature
PreT1
Young Old
IL-6 IP-10 LPS
Young
Control
Day14
Differentiation
-
+
Middle-aged Old Middle-aged(HIV-)D28
8%
26%
29%
37%
7 14 21 28 7 14 21 280
20
40
60
80
100
120
ProT1
ProT2
7 14 21 28 7 14 21 280
20
40
60
80
100
120
ProT2
PreTimmature
7 14 21 28 7 14 21 280
20
40
60
80
100
120
PreTimmature
7 14 21 28 7 14 21 280
20
40
60
80
100
120
PreT1
Differentiation
+
-D28
7%
11%
21%61%
IR NIR
HIV+
B
P < 0.01
r = -0.42
% C
D3
8+
in
CD
8+
me
m T
ce
lls
CD34+ cell count (cell/µl)
0 1 2 3 4 50
20
40
60
80
Pla
sm
a s
CD
14
(p
g/m
l)
CD34+ cell count (cell/µl)
0 1 2 3 4 50
1000
2000
3000
4000
P < 0.0001
r = -0.51P < 0.01
r = -0.42
% C
D3
8+
in
CD
8+
me
m T
ce
lls
CD34+ cell count (cell/µl)
0 1 2 3 4 50
20
40
60
80
Pla
sm
a s
CD
14
(p
g/m
l)
CD34+ cell count (cell/µl)
0 1 2 3 4 50
1000
2000
3000
4000
P < 0.0001
r = -0.51
14 28 14 28 14 28 14 280.0
0.5
1.0
1.5
2.0
2.5
3.0
Control IL-6 IP-10 LPS
* * ns ns
Exp
an
sio
n le
ve
l o
f C
LP
(/ D
7)
Differentiation Altereddifferentiation
A
7 14 21 28 7 14 21 28 7 14 21 280
5
10
15
20
25
30
HIV- HIV+
IR NIR
** ns
*
% o
f C
ells
M
7 14 21 28 7 14 21 28 7 14 21 280
20
40
60
80
100
120
ProT1
ProT2
PreTi
PreT1
HIV- HIV+
IR NIR
% o
f C
ells
M
CLPinduction CLPdistribution
7 14 21 28 7 14 21 280
20
40
60
80
100
120
PreTimmature
Old
B C
Differentiation
+
-
Reduced in vitro TLP induction in the presence of inflammatory molecules
In vitro TLP distribution in the presence of inflammatory cytokines
Ex vivo association between CD34+ cell numbers and inflammation markers
Inflammation => impaired lymphopoiesis = EXTRINSIC FACTOR