Schistosomiasis and HIV co-infection in fishing communities along Lake Victoria, Uganda

Pontiano KaleebuMedical Research Council (MRC UK)

Uganda Virus Research Institute (UVRI)ENTEBBE

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Introduction• A wide geographic overlap in occurrence NTDs and

HIV

• Infection with S.mansoni may be associated with increased susceptibility to HIV infection and disease progression (earlier studies inconclusive)

• Fishing communities in Uganda with high HIV infection rates and high S. mansoni prevalence (50%), suitable for investigation of interactions of HIV and worms 2

Division of labour in the fishing community

3

Project goals

• Impact of S. mansoni on HIV acquisition and associated immune responses in co-infected individuals (Case control study)

• Impact of praziquantel treatment on HIV disease progression and immunological responses among individuals (Intervention study)

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Study areaIn 2009, 1000 HIV- at risk volunteers were enrolled from 5 fishing sites along lake Victoria to assess HIV incidence and study retention.- They were followed six monthly for 18

months- HIV prevalence 29%, incidence

5/100pyo) and high S. mansoni prevalence (50%)

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Case control study Objectives

• To determine the odds of S.mansoni infection among 50 HIV incident cases compared to 150 HIV negative controls identified from a fisher folk cohort

• To compare prevalence of S. mansoni infection status from stored blood samples at enrollment and at 18 months among 50 HIV incident cases and 150 HIV negative controls

• To investigate innate and adaptive immune responses among HIV incident cases with worm infections 6

Study design-nested case control

1 2 3 4 (5)

0 mo 6 mo 12 mo 18 mo

(Recalled volunteers for schistosomiasis Case-control study nested within the prospective cohort

50 HIV+ incident cases150 HIV- controls) were enrolled

Enrol

1000 HIV-1 negative 13-49 yrs at high risk of infection,

HIV VCTPlasmaSerum

HIV VCTPlasmaSerum

HIV VCTPlasmaSerum

HIV VCTPlasmaSerum

HIV VCTPlasmaSerum

Kato Katz on 3 consecutive stool samples to ascertain S.mansoni infection among cases and controls at study exit 35 ml blood :2.5 ml PAXgene tube~30 ml PBMC + plasma0.6 ml whole blood assay

Circulating Anodic Antigen (CAA) test (van Dam / Corstjens) on stored plasma (visits 1,2,3,4)

visit:

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Results

S. Mansoni + S. Mansoni - Total

HIV + cases 26 24 50

HIV - controls

76 74 150

102 98 200

OR 1.05 95%CI (0.53 – 2.10) No evidence of association between S. mansoni infection and HIV sero-conversion

CAA serum analysis is on-going to establish if S.mansoni infection occurred before HIV sero-conversion

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Innate signalling patterns in HIV – Schistosome co-infection

Whole blood:RPMI (1:1) + innate stimuli:

24h

sups

medium

LPS (100 ng/ml) TLR4

PAM3Cys (100 ng/ml) TLR1/2

FSL-1 (50 ng/ml) TLR2/6

Mannan (100 ug/ml)DC-SIGN, MR

LPS + Mannan

CL097 (1 ug/ml) TLR7/8

CpG (5 ug/ml) TLR9

SEA (10 ug/ml)DC-SIGN, MR, xx

LPS + SEA

Curdlan (100 ug/ml) Dectin-1

cytokines (Luminex) TNF-

α

IL-1

0

IL-1

3

IFN

-γ

IFN

-α

Fisher Folk HIV-1 incidence study – schistosomiasis substudy

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Fisher Folk HIV-1 incidence study – schistosomiasis substudyAcquired responses

HIV PTE pools GAG (~1 μg/ml, 24 h, costim + BFA from start) POL

ENVNEFNEG controlSEB

Schistosome antigens SEA(10 μg/ml, 24h*, BFA added AWAfor last 6 h) NEG control

If cells are sufficient:(2nd priority) pos/neg regulators panel[live CD3 CD4 CD8 CD160 LAG-3 PD-1 2B4 CTLA-4](3rd priority) Tregs panel[live CD3 CD4 CD25 CD45RO FOXP3 CD127](4th priority) differentiation panel[live CD3 CD4 CD8 CD27 CD45RA CD127 CCR7](5th priority) cytotoxic panel[live CD8 Grm A Grm B GrmK Pf CD 45RA CD127]

Flow cytometry

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Cytotoxicity genes and Treg associated genes: enhanced in HIV+SM+

Marielle Haks et al. unpublished

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0.2

.4.6

.81

% c

ells

CD4 - ENV

0.5

11.

52

% c

ells

CD8 - GAG

01

23

% c

ells

CD8 - ENV

0.5

11.

5%

cel

ls

CD4 - GAG

+ + +

+ +

+ +

+

+

+

+

+

SM+SM- SM+SM- SM+SM- SM+SM- SM+SM- SM+SM- SM+SM-

+ + +

+ +

+ +

+

+

+

+

+

SM+SM- SM+SM- SM+SM- SM+SM- SM+SM- SM+SM- SM+SM-

TNF-αIL-2IFN-γ

HIV+ S. m.-HIV+ S. m.+

S. mansoni & HIV co-infected individuals have higher Th1 type responses to HIV than HIV mono infected

Andrew Obuku 2012 (unpublished)

P=0.04

P=0.03

P=0.04

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Intervention studyObjectives• Compare HIV progression (viral loads, CD4

count, clinical) among HIV+/S.mansoni+ patients treated with intensive (quarterly) Vs standard (annual) treatment with praziquantel

• Compare immunological changes between the treatment arms over the study period

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Study design- RCT

360 HIV+

226 SM+

113 pzqannually

113 pzqquarterly

134 SM-

follow up 3 monthly

Participant recruitment starting this month and will be followed 3 monthly for 15 months

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Immunological and gene expression profile studies

Innate and adaptive assaysLuminex assaysFlow cytometryRNA expression profiling (RT-MLPA;

Illumina)

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Funders: EDCTP; IAVI; MRC-UK; IDEA (EU)Fisherfolk and Intervention study investigators: P. Kaleebu, A. Kamali, J.Seeley, A. Gershim; L. Nielsen; J Mpendo; A. Elliott; P. Pala ; J Levin; J Nakiyinji-Miiro; G Pantaleo; S. DingSchistosomiasis /HIV Immunology studies: P. Pala; A. Elliott; A. Obuku ; P Kaleebu; R Sekaly; M. Cameron; M. Haks; H. Smits; M. Perrau; A. Harari; S. Ding; G. Pantaleo

Acknowledgements

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