human immuno deficiency virus type 1 (HIV-1) in relation to placental malaria in Yaounde, Cameroon.

In utero mother-to-child transmission (MTCT) of the

Anfumbom KFUTWAH MIM-Yaounde November 2005

Risks factors associated with Mother-To-Child Transmission ( MTCT) of HIV-1, in Yaounde, Cameroon
Pilot study (2000): Nevirapine reduced MTCT of HIV by about 50% Failure to Prevent MTCT of HIV after nevirapine in a pilot study: 1. High maternal viral load (p<0.05) 2. Low birth weight (p<0.01), the female sex (p<0.05) 3. Deliveries 3 months after the peaks of the local rains (p<0.01)
350
PPT MTCT

60

300

50

R=0.634; p<0.001
250 40 200 30

Mm rainfall

150 20 100 10 50 0
mars00 avr00 mai00 juin00 juil00 aot00 sept00 oct00 nov00 dc00 janv01 fvr01 mars01 avr01 mai01 juin01 juil01 aot01 sept01 oct01 nov01 dc01 janv02 fvr02

Role of malaria?

-50

-10

Months

Role of Malaria?

Ayouba et al 2003

MTCT RATE

Malaria, HIV-1 and Placental Environment

P. falciparum infection stimulates the secretion of pro-inflammatory placental cytokines
Role of malaria in seasonal in utero HIV-1 transmission? HIV
LIF RANTES MIP-1 IL-4 IL-10 PROTECTION

P. Falciparum ?
TNF- IL-8 IL-1 IL-6 TRANSMISSION

Modifications of the placental environment by co-infections (P. falciparum) could play an important role in the in utero transmission of HIV-1

Hypothesis

Methods (1)
81HIV-1+ 50HIV-1-

Maternal blood

Placental Fragments

Thin and thick blood smears Blood count CD4 Viral Load HBV, HCV, Syphilis

Malaria

Cytokine profiles

Results (1) : HIV-1 and Malaria 1.

40 35 30 25 20 10 0

HIV-1 Positive HIV-1 Negative 14/50

15/78
Periphery

Malaria Positive [%]

15 5
16

9/48 9/48 8/45

Placenta

2.
Parasitemia

14

10 8 6 4 2 0

13 9 4 1
HIV positive

<1% parasitemia 1-2.5% parasitemia 2.5-5% parasitemia 8

4 0 0
HIV negative

HIV positive

HIV negative

Periphery

Placenta

3.

Reciprocal discrepancies of malaria infection between placenta/periphery (4/7: >50%)

4. Tendency of higher MTCT of HIV-1 in women with co-infections

Placenta Infections in vitro : TNF alpha and the histoculture system

A recently validated system (placental histoculture) : culture up to 10 days.

Previous studies
Cells of placental origin are resistant to cell-free viruses

9 HIV-1 genome bearing the envelope protein G of Vesicular Stomatitis

Virus (VSV-G) by-pass this resistance.

9Effects of TNF alpha on HIV infected placenta?

Methods (2): Histoculture (Luminometer and Imagery)

Extensive washing of placental fragments

1
Overnight infection with HIV-1(VSV-G) (0,2ng and 2ng) per fragment Luminometer

Luminescence in tissue lysate

1. 2. 3.

No TNF-alpha 5ng/ml of TNF-alpha 50ng/ml of TNF alpha

48hrs, 96hrs, 120hrs

2
A Faye et al Placenta 2005

Imagery

Xenogen IVIS(tm) and Living Image(r) Software.

Results (2): Histoculture (Luminometer and Imagery) TNF alpha significantly (p < 0.001) increases viral replication in the placenta
12

Ratio of infection

HIV-1(VSV-G) 0.2 ng/fragment

8

1 #1
TNF-alpha 0 ng/ml 5 ng/ml 50 ng/ml

3 #2

#3
0 48h 96h 120h

2
Time post-infection (hours)

#4 4

HIV (VSVG) #1: no TNF alpha 31501 # 2: 5ng/ml TNF alpha 93449 HIV envelope deficient (controls) #3: no TNF alpha #4: 5ng/ml alpha TNF

Preliminary Results: Co-infections (Plasmodium & HIV) in the Histoculture System Method Infect placenta fragments with HIV (VSV-G) Fragments in contact with iRBC and uRBCs in Shaker for 1 hour
to about 50% in Plasmagel
Cintique VSV-G/Pf.
4E+4

0,2 ng VSV-G/ 2x10 4

uRBC iRBC

3E+4

2E+4

1E+4

0E+0 J2 J3 J4 J5 Jours Post Pf Days post parasite introduction

Cintique VSV-G/Pf.
3E+5 2E+5 2E+5

2 ng VSV-G/ 2x10 4

-iRBCs (group O+) concentrated

uRBC iRBC

Histoculture Variability RBC

1E+5 5E+4 0E+0 J2 J3 J4 J5

Days post parasite introduction Jours post Pf

Conclusions
Cell free virus infection Cell to cell Infection Hofbauer cells X ?

Surface molecules

Activation by Coinfections (malaria)

TNF-alpha increases HIV-1 replication in the human placenta Increase of HIV-1 MTCT ? Decrease the efficiency of preventive strategies ?

HIV-1 MTCT and malaria prophylaxis adjusted according to the local environment

Collaborations
National
Centre Pasteur, Cameroun (A. Ayouba, M. Tejiokem, B. Njinku, E. Nerrienet, D. Rousset) Maternit de CASS Nkoldongo-Yaound Maternit de lHpital Central- Yaound Fondation Chantal Biya- Yaound

International
Unit de Rgulations des infections Rtrovirales, IPP (S. Pornprasert, A.Faye, G Dolcini, F BarrSinoussi, E Menu) Unit dimmunologie Molculaire des Parasites, IPP (Charlotte Behr, S loizon, O puijalon) INSERM U131 (G.Chaouat) INSERM U717 (Jean Yves Mary)

European Network for the study of in utero transmission of HIV-1 (Biomed)