Transcript HIV persistence in the testis?

2014 “Towards an HIV Cure” symposium
Melbourne
SIV infection of the macaque testis –
an immune privileged site
Wendy Winnall, University of Melbourne
HIV persistence in the testis?
Does HIV persist in the testis?
Why should we care?
•
•
Transmission through seminal plasma?
?
Reservoir during cART?
Is the testis a sanctuary for HIV?
Possible mechanisms for HIV persistence in
the testis:
•
Blood testis barrier?
•
Testicular immune privilege?
?
Cells of the testis
Interstitium
Seminiferous tubule
Lumen
lymphatic
vessel
blood
vessel
Cells of the testis
Interstitium
Seminiferous tubule
Lumen
Spermatids
lymphatic
vessel
blood
vessel
Spermatocytes
Spermatogonia
Cells of the testis
Interstitium
Seminiferous tubule
Sertoli cells
Lumen
lymphatic
vessel
blood
vessel
Cells of the testis
Interstitium
Seminiferous tubule
Sertoli cells
Blood-testis
barrier
Lumen
Pgp
lymphatic
vessel
blood
vessel
Cells of the testis
Interstitium
Seminiferous tubule
Sertoli cells
Blood-testis
barrier
Lumen
Pgp
Leukocytes:
macrophages
T cells
lymphatic
vessel
blood
vessel
Leydig cells
Is the testis a sanctuary for HIV?
Possible mechanisms for HIV persistence in
the testis:
•
Blood testis barrier?
•
Testicular immune privilege?
?
Is the testis a sanctuary for HIV?
Possible mechanisms for HIV persistence in
the testis:
•
Blood testis barrier?
•
Testicular immune privilege?
•
•
Suppression of innate and acquired immune
responses in the testis of rodents
Predicted to protect immunogenic spermatids
from immune responses and inflammation
?
Pigtailed macaque model for HIV
infection
SIV infects the pigtailed macaque testis
Shehu-Xhilaga et al Retrovirology, 2007
Aims
•
•
•
Are immune responses suppressed in the
macaque testis?
How does SIV infection affect the testis?
Do testicular CD8 T cells respond to SIV
infection?
SIV infection of pigtailed macaques
Testes and blood taken from:
•
7 uninfected macaques
•
4 macaques infected with SIVmac239
•
•
10 weeks
24 to 30 weeks
Is there immunosuppression in the
macaque testis?
unstimulated
mitogen stimulation
Is there immunosuppression in the
macaque testis?
unstimulated
mitogen stimulation
Wilcoxon Signed Rank test
•
T cell responses in the testis are lower than in the blood
Aims
•
•
•
Are immune responses suppressed in the
macaque testis?
How does SIV infection affect the testis?
Do testicular CD8 T cells respond to SIV
infection?
SIV damages the macaques testis
uninfected
SIV week 10
VERALL T and B.jo
Layout: fig for poster
SIV alters testis leukocyte populations
uninfected testis CD3+
105
104
40.2
SIV week 10 CD3+
10
5
10
4
10
CD8
CD8
83
103
3
41
/7/14 1:09 PM
3.77
0
0
0
10
3
10
CD4
4
10
5
0
10
3
10
4
10
5
CD4
Page 1 of 1
(FlowJo v9.7.5)
VERALL T and B.jo
Layout: fig for poster
SIV alters testis leukocyte populations
uninfected testis CD3+
CD4 cells in the testis and blood
SIV week 10 CD3+
103
41
/7/14 1:09 PM
3.77
0
10
5
0
10
3
10
4
10
5
CD4
0
k3
0
k1
w
4
CD8 cells in the testis and blood
100
blood
WB
testis
testis
80
60
40
20
0
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0
10
CD4
w
3
k3
10
w
0
10
0
% CD8 out of CD3 cells
0
20
0
3
30
k1
10
CD8
CD8
83
40
w
10
4
WB
blood
testis
testis
un
in
fe
ct
ed
40.2
5
un
in
fe
ct
ed
104
10
% CD4 out of CD3 cells
50
105
(FlowJo v9.7.5)
VERALL T and B.jo
Layout: fig for poster
SIV alters testis leukocyte populations
uninfected testis CD3+
CD4 cells in the testis and blood
SIV week 10 CD3+
103
41
3.77
0
0
10
3
10
4
10
5
10
0
CD4
10
3
10
4
10
5
CD4
0
0
k3
0
20
w
3
30
0
10
CD8
CD8
83
40
k1
10
4
WB
blood
testis
testis
w
40.2
5
un
in
fe
ct
ed
104
10
% CD4 out of CD3 cells
50
105
CD8 cells in the testis and blood
/7/14 1:09 PM
20
k3
0
0
w
Shift towards effector memory and
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away from central memory
40
0
•
60
k1
Influx of CD8 T cells
80
w
Loss of CD4 T cells
blood
WB
testis
testis
un
in
fe
ct
ed
•
•
100
% CD8 out of CD3 cells
SIV infection lead to:
(FlowJo v9.7.5)
Aims
•
•
•
Are immune responses suppressed in the
macaque testis?
How does SIV infection affect the testis?
Do testicular CD8 T cells respond to SIV
infection?
Are there CD8 T cell responses in the
SIV-infected testis?
mer.jo
uninfected testis CD8+
5
10
0.0354
104
10
tetramer
KP9-ManeA*084
KP9 tetramer
tetramer
KP9-ManeA*084
KP9 tetramer
10
SIV week 10 CD8+
5
3.93
104
3
10
102
3
102
0
0
10
3
CD8
10
4
10
5
0
0
10
3
CD8
10
4
10
5
Are there CD8 T cell responses in the
SIV-infected testis?
mer.jo
102
0
0
10
3
CD8
•
10
4
10
5
2
0
0
0
10
3
10
4
10
5
0
102
3
4
k3
10
testis
testis
w
3
blood
WB
0
10
3.93
104
6
k1
0.0354
104
5
w
10
SIV-specific CD8 T cells in testis and blood
KP9 tetramer positive cells in blood and testis
% KP9+ out of CD8+ cells
5
tetramer
KP9-ManeA*084
KP9 tetramer
tetramer
KP9-ManeA*084
KP9 tetramer
10
SIV week 10 CD8+
un
in
fe
ct
ed
uninfected testis CD8+
CD8
SIV-specific CD8 T cells were detected in the testis
SIV inhibits CD8 T cell cytokine
production
layout for poster
SIV wk 10 testis CD8+
10
5
SIV wk 10 blood CD8+
0.0831
0
10
0.365
1.99
104
TNF
TNF
104
5
10
3
3
10
0
0
97.7
2.2
0
3
10
10
4
10
91.8
5
5.88
0
IFN
3
10
10
4
10
5
IFN
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(FlowJo v9.7.5)
SIV inhibits CD8 T cell cytokine
production
layout for poster
3
10
0
0
97.7
2.2
0
3
10
10
4
10
91.8
5
0
IFN
•
5.88
3
10
10
IFN
4
10
5
WB
blood
testis
testis
10
5
0
0
3
15
k3
1.99
w
10
0.365
104
TNF
TNF
104
5
10
0
0
k1
0.0831
w
5
un
in
fe
ct
ed
10
Cytokineproduction
productionafter
by CD8
cells stimulation
Cytokine
mitogen
SIV wk 10 blood CD8+
% CD8+ producing IFNg or TNF
SIV wk 10 testis CD8+
SIV infection inhibited cytokine production by CD8 T
cells in response to mitogens
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(FlowJo v9.7.5)
Summary
•
T cell responses to mitogens are relatively low in
the macaque testis
Evidence for immune privilege in the primate testis
•
SIV infection affects the macaque testis:
•
Damage to the testis architecture
•
Influx of effector memory CD8 T cells
•
Loss of CD4 T cells
Summary
•
•
SIV-specific CD8 T cells were detected in the testis
However cytokine production in response to
mitogens was inhibited by SIV infection
Immunosuppression in the macaque testis reduces
the ability of CD8 T cells to respond to infection,
making the testis vulnerable to persistent SIV infection
and damage.
•
Testicular immune privilege
Acknowledgements
Kent Lab, Doherty Institute
University of Melbourne
Stephen Kent
Rob De Rose
Sarah Lloyd
Vinca Alcantara
Thakshila Amarasena
Marit Kramski
Shayarana Gooneratne
Hyon-Xhi Tan
Monash University
Mark Hedger
Dept of Obstetrics and
Gynaecology,
University of Melbourne
Jane Girling
CSIRO AAHL Geelong
Megan Schepers
Lisa Reid