Presented at the 6th Int. Workshop on HIV Transmission, 14 – 15 July 2011, Rome, Italy

HIV-1 and Reactivated Coinfecting Herpesviruses Rearrange the Cytokine Network in Semen

of Infected Individuals

Andrea Introini,

National Institute of Child Health and Human Development

NIH

Bethesda, MD, U.S.A.

Presented at the 6th Int. Workshop on HIV Transmission, 14 – 15 July 2011, Rome, Italy

SEMEN

• Is a complex fluid that contains spermatozoa and

soluble factors, including cytokines

• Natural carrier of many pathogens

• Pathogens replication is affected by cytokines and

also affect cytokine expression

Do HIV-1 infection and coinfecting herpesviruses

affect cytokine profile in semen?

Presented at the 6th Int. Workshop on HIV Transmission, 14 – 15 July 2011, Rome, Italy

OUR STUDY

• Viral load of:

HIV-1

Herpes Simplex Virus-2 (HHV-2)

Epstein Barr Virus (HHV-4)

Cytomegalovirus (HHV-5)

HHV-6 A and B

HHV-7

HHV-8

Blood and seminal plasma from:

• 74 HIV-1-infected individuals therapy naïve

• 33 uninfected individuals

• Cytokines

Presented at the 6th Int. Workshop on HIV Transmission, 14 – 15 July 2011, Rome, Italy

HIV-1 LOAD L

og

10 (

co

pie

s/m

l)

Blood Semen

100%

positive

94%

positive

HIV-1 blood load Log10 (copies/ml)

HIV

-1 s

em

en load

Log

10 (

co

pie

s/m

l)

R=0.64 p<0.0001

4.5 4.0

Presented at the 6th Int. Workshop on HIV Transmission, 14 – 15 July 2011, Rome, Italy

0

20

40

60

80

HERPESVIRUSES REACTIVATION

p<0.001

HIV-1-infected HIV-1-uninfected

Semen

Blood

po

sitiv

e s

am

ple

s (

%)

CMV ML vs log10(copies/ml)

* *

positiv

e s

am

ple

s (

%)

EBV ML vs log10(copies/ml)

0

5

10

Presented at the 6th Int. Workshop on HIV Transmission, 14 – 15 July 2011, Rome, Italy

1

2

3

4

5

6

7

8

9

CMV

Vira

l lo

ad

Log

10

(copie

s/m

l)

Blood Semen

HIV-1-CMV coinfected

77% semen only

19% both semen and blood

3% blood only

CMV REACTIVATION IS COMPARTMENTALIZED

Presented at the 6th Int. Workshop on HIV Transmission, 14 – 15 July 2011, Rome, Italy

CONCURRENT EBV AND CMV INFECTION IS ASSOCIATED WITH AN

HIGHER HIV-1 SHEDDING IN SEMEN

p<0.05 V

ira

l lo

ad

Log

10 (

copie

s/m

l)

HIV-1 *

Presented at the 6th Int. Workshop on HIV Transmission, 14 – 15 July 2011, Rome, Italy

CYTOKINE PANEL

• Pro-inflammatory cytokines: IL-1α, IL-1β, IL-6,

IFN-γ, TNF-α

• Anti-inflammatory cytokines: TGF-β

• CCR5 ligands: RANTES, MIP-1α, MIP-1β, Eotaxin

• CXCR4 ligands: SDF-1β

• Chemokines: MCP-1, MIP-3α, IP-10, MIG, IL-8

• Growth factors: IL-2, IL-7, IL-15, IL-16, GM-CSF

Presented at the 6th Int. Workshop on HIV Transmission, 14 – 15 July 2011, Rome, Italy

SEMEN AND BLOOD ARE SEPARATE IMMUNE COMPARTMENTS (UNINFECTED MEN)

Semen/Blood MEDIAN RATIO

Enriched

in semen

Enriched

in blood

*

* *

* *

* * *

*

*

* *

*

= p<0.05 *

Presented at the 6th Int. Workshop on HIV Transmission, 14 – 15 July 2011, Rome, Italy

HIV-1 UPREGULATES FEW CYTOKINES IN BLOOD

N-f

old

0

2

4

6

8

HIV-1+/HIV-1- RATIO

0.1

= p<0.05

1

* * *

* *

*

*

*

Presented at the 6th Int. Workshop on HIV Transmission, 14 – 15 July 2011, Rome, Italy

1

3

5

7

N-f

old

= p<0.05

HIV-1 UPREGULATES MANY CYTOKINES IN SEMEN

HIV-1+/HIV-1- RATIO

* *

*

*

* * *

*

*

*

* * * *

*

*

*

Presented at the 6th Int. Workshop on HIV Transmission, 14 – 15 July 2011, Rome, Italy

HIV-1 INFECTION REARRANGES THE CYTOKINE NETWORK IN SEMEN

HIV-1-infected

HIV-1-noninfected

Spearman’s rank

correlation coefficient

Presented at the 6th Int. Workshop on HIV Transmission, 14 – 15 July 2011, Rome, Italy

CONCLUSIONS

• EBV and CMV reactivate in the course of HIV-1 infection

predominantly in semen

• HIV-1 infection increases the number and the strength of

correlations between cytokines in semen

• Concurrent shedding of CMV and EBV in semen is

associated with higher HIV-1 seminal load

• HIV-1 infection upregulates more cytokines in semen than

in blood

• Blood and semen are two separate immunological

compartments with different cytokine profiles

Presented at the 6th Int. Workshop on HIV Transmission, 14 – 15 July 2011, Rome, Italy

CONCLUSION

HIV-1 together with its copathogens significantly change the

immunological landscape of semen, probably affecting HIV-1

evolution in this compartment and selecting for transmissible

HIV-1 variants.

Presented at the 6th Int. Workshop on HIV Transmission, 14 – 15 July 2011, Rome, Italy

Leonid Margolis

Andrea Lisco

Christophe Vanpouille

Jean-Charles Grivel

Wendy Fitzgerald

Anush Arakelyan

Sarman Singh

Arshi Munawwar