DEPARTEMENT MICROBIOLOGIE

 
INHOUD

1. Inleiding
2. Focus on
PROJECTS
Publications in international peer-reviewed journals
Other publications and abstracts

1. Inleiding
Het hoofddoel van het Departement Microbiologie is het verbeteren van de kennis en de bestrijding van het Humaan Immuundeficiëntie-Virus (HIV), tuberculose (TB) en seksueel overdraagbare aandoeningen (SOA) in ontwikkelingslanden. Het bestaat uit de vier eenheden Virologie, Immunologie, Mycobacteriologie en SOA/HIV Research & Interventie. Het Departement als geheel en de verschillende eenheden apart kregen een zeer goede beoordeling van de externe audits en de Wetenschappelijke Raad van Advies eind 2001. De onderzoeksteams gaan dus met vernieuwde moed en zonder grote wijzigingen verder op de ingeslagen weg. Ze hopen wel dat deze evaluatie de overheden ervan zal overtuigen hun werk eindelijk afdoende te ondersteunen.
De belangrijkste onderzoekslijnen zijn de moleculaire variabiliteit van het HIV en de implicaties voor vaccinontwikkeling, therapie en diagnose; epidemiologisch en immunologisch onderzoek naar HIV en de interacties met andere infecties; de preventie, diagnose, behandeling en bestrijding van SOA en HIV in ontwikkelingslanden; de ontwikkeling en epidemiologische toepassing van diagnostica voor tuberculose en andere mycobacteriële aandoeningen, en medicamenteuze resistentie.
Het onderwijs concentreert zich op postuniversitaire professionele en wetenschappelijke opleidingen in de microbiologie, immunologie en ziektebestrijding, aan het ITG maar ook in universitaire en internationale programma’s (zie hoofdstuk Onderwijs).
De dienstverlening omvat nationale en internationale diagnostische referentietaken, en de ondersteuning van instellingen en bestrijdingsprogramma’s in de ontwikkelingslanden (zie hoofdstuk Ontwikkelingssamenwerking).

Administration of the Microbiology Department: a solid team.

De Eenheid Virologie heeft het voorbije jaar verder gesleuteld aan de optimalisatie van in vitro-testen voor het meten van HIV-neutralisatie. Deze testen zijn niet alleen van belang voor het basisonderzoek maar ook voor de toekomstige evaluatie van anti-HIV-vaccins. De Eenheid ontwikkelde ook een gelijkaardige test voor de kwantitatieve bepaling van neutraliserende antilichamen met gebruik van niet-infectieuze viruspartikels of zogenaamde pseudo-virussen. Hierdoor is het niet meer strikt noodzakelijk om dergelijke testen uit te voeren in hoogbeveiligde L3-laboratoria en kunnen ze op termijn hopelijk hun toepassing vinden in ontwikkelingslanden, waar de meeste vaccintrials zullen plaatsgrijpen. Ten slotte ontwikkelde de Eenheid ook een test voor de grootschalige genetische karakterisatie van de HIV-1 Circulerende Recombinante Vorm CRF02_AG, die predominant aanwezig is in West- en Centraal-West-Afrika. Deze test kan van belang zijn voor de evaluatie van kandidaat HIV-1-vaccins tegen dit specifieke subtype.
Eind 2002 ging Guido van der Groen, vele jaren leider en bezieler van de Eenheid Virologie, met emeritaat. Een waardige opvolger vinden is alvast de grote uitdaging voor het volgende jaar.

The Amplispot method. The Unit of Immunology and the “Gladstone Institute of Virology and Immunology”, UCSF, San Francisco, together developed a sensitive method for measuring cellular immune responses. The sensitivity of existing ELISPOT-methods is sharply increased by the addition of recombinant cytokines (IL-7 and IL-15).
This method could be useful in the follow-up of cellular immune response after vaccination with experimental (e.g. anti-HIV) vaccines .
A. The ELISPOT method without antigen. B. The ELISPOT method with PPD (protein purified derivative, an antigen derived from Mycobacterium tuberculosis). C. The Amplispot method without antigen. D. The Amplispot method with PPD. E. Summary graph.

De Eenheid Immunologie breidde in 2002 haar onderzoek in de cellulaire immunologie van HIV uit met projecten rond virale fitheid, immuuntherapie van HIV en in vitro-evaluatie van experimentele microbiciden. Het onderzoeksproject in Ivoorkust over de immunologie van HIV ondervond wel wat hinder van de politieke instabiliteit. Het onderzoek op HIV en het verwante Simian Immune Deficiency Virus (SIV) in chimpansees, in samenwerking met het primatencentrum in Rijswijk, Nederland, werd na tien jaar afgerond. Dit sociaal omstreden onderzoek bij primaten zal in de toekomst waarschijnlijk niet meer mogelijk zijn. Dank zij het AIDS-Impulsprogramma (zie hoofdstuk Ontwikkelingssamenwerking) kon de Eenheid aanzienlijk bijdragen tot de capaciteitsversterking van het laboratorium voor virologie en bacteriologie van het Centre Hospitalier Universitaire (Prof. S. Mboup) te Dakar, Senegal, en van het laboratorium voor immunologie van het Instituto de Medicina Tropical (Prof. E. Gotuzzo, Prof. J. Arevalho) te Lima, Peru. Verder verzorgden we opleidingen voor wetenschappers en technici voor andere projecten van het AIDS-Impulsprogramma (CHKU, Rwanda; PNLS, Kinshasa). Nu antiretrovirale behandeling ook in ontwikkelingslanden ingang vindt, neemt de vraag naar degelijke laboratoriumopleidingen sterk toe, onder meer voor de bepaling van CD4 T-cellen.


The Unit of Immunology developed an in-vitro laboratory model for testing new antiviral drugs for prevention of HIV. In this model, human dendritic cells (DC) and T cells (T4) are used. Both cell types are present in humans and play a crucial role during sexual HIV transmission. The HIV virus is pre-treated with the drug and afterwards used to infect DC. The infected DCs are then co-cultured with T cells with or without the addition of the antiviral drug. After 2 weeks the effect of the antiviral product on the HIV virus is analysed. Drugs which completely inhibit the virus are selected and further investigated.

Hôpital Le Dantec, CHU, Dakar, Senegal: from left to right: Luc Kestens, Chris Vereecken and Tandakha Dieye. A project of laboratory capacity strengthening.

De Eenheid SOA/HIV Research en Interventie startte een nieuw project in Kisumu, Kenia voor de preventie van HIV en andere reproductieve gezondheidsproblemen bij jongeren, als een logisch vervolg op het eerdere epidemiologische onderzoek naar HIV en het seksueel gedrag van jongeren in deze streek. De resultaten van het kwalitatieve onderzoeksluik werden zeer gunstig onthaald in lokale workshops, die we in 2003 zullen uitbreiden naar de drie andere steden waar dit vergelijkend onderzoek plaatsvond, namelijk Ndola in Zambia, Cotonou in Benin en Yaoundé in Kameroen.
Het SOA-laboratorium spitste zich in 2002 vooral toe op het ontwikkelen van de moleculaire differentiatie van trichomonas, een parasitaire SOA-verwekker. Deze test zal nu zijn toepassing vinden in een studie naar de hoge prevalenties van trichomonas bij jonge, nog niet seksueel actieve vrouwen in Zambia. In samenwerking met het Institut National de Santé Publique in Abidjan en de London School of Hygiene and Tropical Medicine startten we een nieuwe studie in Ivoorkust over de preventie van moeder-kindtransmissie van HIV in obstetrische diensten.

Yvan Best, a young Peruvian scientist working on the flow cytometer.

Een pas goedgekeurd project (ORISS, Operational Research in STI and Related Services for Women in High Risk Situations in Cambodia and Thailand) zal algoritmes voor de behandeling van SOA bij prostituees valideren, en de sociale achtergrond, mobiliteit en noden aan gezondheidszorg van deze doelgroep bestuderen.
De Eenheid vernieuwde ook haar contract met Family Health International voor het leveren van technische ondersteuning aan projecten van de Amerikaanse internationale samenwerking (IMPACT), vooral voor de diagnose en behandeling van SOA, en de antivirale behandeling van HIV.
We zetten ook de institutionele samenwerkingsverbanden voort in Ivoorkust (Institut National de Santé Publique) en in Cambodja (National STD/HIV Control Programme). Het AIDS-Impulsprogramma gaf een aanzienlijke bijkomende ondersteuning aan de lokale capaciteitsversterking, onder meer via investeringen in infrastructuur, opleidingen en technische ondersteuning. Dank zij het AIDS-Impulsprogramma konden we ook capaciteitsversterking en onderzoek aanvatten in Kinshasa (D.R. Congo). In samenwerking met het Programme National de Lutte contre le SIDA (PNLS) en Artsen Zonder Grenzen voerden we een enquête uit naar het gebruik van antiretrovirale middelen in Kinshasa, met een verlengstuk in Pointe-Noire (Congo-Brazzaville). Tien jaar na onze eerdere studies herhaalden we ook een HIV/SOA-enquête bij prostituees in de Matonge-kliniek in Kinshasa, en vulden deze aan met een gelijkaardige studie in een andere SOA-kliniek, het Centre Victoire.
De Eenheid organiseerde in mei 2002 het tweede wereldcongres over microbiciden, dat werd bijgewoond door meer dan 600 deelnemers van alle continenten (zie Focus on). Dit gaf een nieuw elan aan ons eigen microbicidenonderzoek, waarvoor drie klinische trials (fase 1) in voorbereiding zijn.

Luc Boel (second from the left) introduced an alternative and affordable CD4 T-cell counting method in the “Centre Hospitalier de Kigali”, Rwanda in 2002 with the support of the AIDS Impulse Programme.

De Eenheid Mycobacteriologie heeft zich, als spil van een internationaal netwerk, verder ingezet voor het volgen van de wereldwijde verspreiding van resistente tuberculose tegen eerste- en tweedelijnsgeneesmiddelen. De resultaten vertonen sterke regionale verschillen. Naast economische oorzaken blijken ook de efficiëntie van de bestrijdingsprogramma’s en het (soms chaotische) voorschrijfgedrag belangrijke factoren in de verspreiding van resistente TB. De Eenheid werkt ook aan de standaardisatie en vereenvoudiging van de technieken voor de bepaling van gevoeligheid voor geneesmiddelen, en de evaluatie van gestandaardiseerde schema’s voor de (her)behandeling van MDR-TB patiënten.
De intense samenwerking bij het onderzoek en de bestrijding van Buruli ulcus en tuberculose in Benin werd in 2002 bekroond met de oprichting van het ‘Laboratoire National de Référence de Mycobactériologie’ in Cotonou. De belangrijkste conclusie van het onderzoek naar Buruli ulcus in 2002 is dat een groot aantal gevallen voorkomt als een niet-ulcererende vorm, met osteomyelitis als de belangrijkste. Het blijkt ook dat het BCG-vaccin tegen tuberculose bescherming kan bieden tegen de ernstigere vormen van Buruli ulcus. In de zoektocht naar het reservoir van Mycobacterium ulcerans verkregen we verder inzicht in de rol van waterinsecten.

The members of the Aids Reference Laboratory: a highly motivatedteam with a strong emphasis on quality.

Het Departement was ook in 2002 zeer actief in de institutionele onderwijsprogramma’s, vooral in de Master of Disease Control (zie hoofdstuk Onderwijs). Samen met het Departement Parasitologie bouwden we ook mee aan de nieuwe oriëntatierichting Tropische Biomedische Wetenschappen aan de Universiteit Antwerpen, en droegen we bij aan de internationale masterprogramma’s van de Vrije Universiteit Brussel.

General Meeting of EC Concerted Action Project: “Improved diagnosis, drug resistance detection, and control of tuberculosis in Latin America”. The First General Meeting of this project was held at the ITM from 1-4 July 2002 and coordinated by Prof. Françoise Portaels.


2. Focus on: Microbiciden 2002, 12-15 mei, Antwerpen



Onze Eenheid HIV/SOA Research en Interventie viel in 2002 de eer te beurt om het tweede congres over Microbiciden te organiseren. Het eerste congres vond plaats te Washington in 2000. Voorzitter van het congres was Marie Laga. De wetenschappelijke coördinatie werd geleid door Lut Van Damme in samenwerking met verscheidene internationale Instituten. Het administratief en technisch personeel van de Eenheid onder leiding van Jan Vielfont en Yvette Baeten zorgde voor een geslaagde organisatie in het Vlaamse Congres en Concertgebouw van Antwerpen. De coördinatie van het secretariaat werd verzorgd door Yvette Jacob.

Zena Stein during the Microbicides 2002 conference in Antwerp.

Het congres werd geopend op 13 mei 2002 door Arnd Hoeveler (afgevaardigde EU), Peter Piot (Directeur UNAIDS, Zwitserland), Kim Dickson-Tetteh (Chris Hani Baragwanath Hospital, Zuid-Afrika) en Charlotte Watts (London School of Hygiene and Tropical medicine, VK). De opening werd gevolgd door een huldiging van Zena Stein door Helen Rees (University of Witwatersrand, Zuid-Afrika). We waren zeer vereerd om deze prille voorvechtster van het concept microbiciden te mogen verwelkomen en te bedanken voor haar jarenlange actieve inzet. Zena Stein is Professor in de Epidemiologie en Volksgezondheid aan de Universiteit van Colombia, New York. Zij blijft ook nu, aan het einde van haar professionele carrière, een voorbeeld voor alle wetenschappers in het microbicidenonderzoek. De heer Eddy Boutmans (Belgisch Staatssecretaris voor Ontwikkelingssamenwerking) sloot het congres af op 15 mei 2002.

Participants at the Microbicides 2002 conference.

Gedurende deze drie dagen werd de topic microbiciden vanuit verschillende invalshoeken toegelicht en geëvalueerd. Meer dan 600 wetenschappers, waarvan meer dan de helft van Afrika en Azië, van basiswetenschappen tot filosofie en economie en komende uit 42 verschillende landen namen deel aan het debat. Het belang van microbiciden voor de emancipatie van vrouwen in ontwikkelingslanden, het ontwikkelen van vertrouwen, respect, assertiviteit en betere communicatietechnieken werd onderstreept. Resultaten van preklinische en klinische studies werden voorgesteld en besproken. Nieuwe producten en evaluatiemethoden werden doorgelicht. De ethiek omtrent het uitvoeren van klinische studies was een belangrijk item op de agenda. Een vroege dialoog werd noodzakelijk geacht om alle partners te betrekken in het vinden van een consensus omtrent het opzetten van de studies. Microbiciden werden ook grondig geëvalueerd vanuit een farmaco-economisch standpunt. Er werd duidelijk naar voor gebracht dat er niet alleen een markt voor deze producten bestaat maar dat zij ook een belangrijk voordeel kunnen bieden op het gebied van volksgezondheid voor de ontwikkelingslanden.
Naast het volgen van de wetenschappelijke presentaties was het congres een zeer belangrijke ontmoetingsplaats en werden vele nieuwe contacten gelegd voor toekomstige samenwerkingsprojecten. Het ‘International Partnership for Microbicides’ werd officieel voorgesteld. Deze groep zal alles in het werk stellen om de ontdekking, ontwikkeling en bereikbaarheid voor vrouwen van een veilig en effectief microbicide te stimuleren. Verder werden er ook 2 symposia gehouden door de National Institutes of Health (NIH, USA), Family Health International (FHI, USA) en de Rockefeller Foundation (USA).

Zena Stein addressing the participants at the Microbicides 2002 conference.

Het congres bracht ons weer een stapje dichter bij een effectief microbicide en heeft duidelijk gemaakt dat nu ook de topic microbiciden op de politieke agenda staat en verdient te staan.
Voor de volledige besluiten en presentaties zie onze ITG-website:
http://www.itg.be/micro2002/


PROJECTS

UNIT OF VIROLOGY

Unit of Virology: L3, the place to be.

Study of the in vitro neutralisation of primary HIV-1 isolates by antibodies induced by vaccination and natural infection
The hypothesis is that there are two sets of epitopes on human immunodeficiency viruses: one which is exposed on the surface of the free virion, and the other which is more readily available to antibodies after the virus has bound to its target cell. The interaction of antibodies with the epitopes gives two different responses, which may be relevant for natural infection and vaccination. When free virions combine with neutralising antibodies, they lose their infectivity. By contrast, when antibodies bind to a cell-associated virus, the virus can still replicate, albeit at a slower rate. For a vaccine, antibodies which produce an all-or-nothing loss in viral infectivity are the ideal, since they can reasonably be expected to prevent the viremic phase following infection, irrespective of the route by which the virus enters the body. However, antibodies which can slow down virus replication may be sufficient for protection following heterosexual transmission, if they can delay the viremic phase until the host’s own cell-mediated immunity can eliminate the virus. An advantage of the latter is that the epitopes, which are revealed when the virus binds to the cell, are present on most virus isolates, while those on the free virion appear to be restricted within viral subtypes.
We are developing assays to quantify neutralising antibodies to the different sets of epitopes. These assays are being applied to plasma from monkeys and human volunteers from vaccine trials. Previously, it had not been possible to demonstrate neutralisation with the viruses most likely to be involved in transmission events using antibodies induced by candidate HIV vaccines. However, we have now demonstrated these antibodies, indicating that the first generation of HIV vaccines are potentially protective in human trials.
ITM promoter: G. van der Groen
ITM collaborators: D. Davis and H. Donners
External collaborators: J. Heeney (Biomedical Primate Research Centre, Rijswijk, The Netherlands); S. Barnett (The Chiron Corporation, California, USA); G. Voss (GlaxoSmithKline (GSK), Rixensart, Belgium); Aventis Pasteur, Pennsylvania, USA and Lyon, France; HIV Vaccine Trials Network, USA
Support: Fund for Scientific Research - Flanders (FWO)

Multi-component HIV vaccines using engineered envelopes: prime-boost strategies using ISCOMs
Previously we have demonstrated protection of rhesus macaques against infection with a recombinant form of simian immunodeficiency virus displaying HIV envelope glycoproteins on their surface (SHIV). We used a recombinant prime, peptide boost vaccination strategy, priming the monkeys’ antibody responses with recombinant HIV envelope glycoprotein and then boosting with peptides representing linear epitopes. The recombinant immunogens expose the monkey’s lymphocytes to the immunogen’s natural conformation, while the peptides focus the response onto the most important epitopes. Two epitopes were selected on the basis of our preliminary studies investigating the neutralising antibody responses of rats to synthetic peptides. However, targeting linear epitopes gives a very limited cross-neutralisation, and it would be better to have three epitopes, so that it is less likely that escape variants will emerge. Our objective is to broaden protection by boosting antibodies to conformational epitopes which are shared by many HIV isolates. Our collaborators in Italy have used phage display technology to select peptides which can specifically bind to antibodies able to neutralize the type of virus involved in natural transmission events. In the new project, these mimotopes will replace the peptides representing linear epitopes: the monkeys will be challenged by the natural transmission route, with SHIV recombinants having envelope glycoproteins from more pathogenic HIV isolates which are also resistant to neutralisation.
Promoters: J. Heeney, S. Barnett
ITM collaborators: G. van der Groen, D. Davis
External collaborators: J. Heeney (Biomedical Primate Research Centre, Rijswijk, The Netherlands); S. Barnett (Chiron Corporation, California, USA); B. Morein (ISCONOVA AB, Uppsala, Sweden); G. Scala (University Federico II, Naples, Italy)
Support: HIVRAD (HIV Vaccine Research and Design), National Institutes of Health, Washington (USA).

Study of HIV variability - Implications for vaccine development:
The epidemiology of HIV-1 subtypes and CRFs is characterised by their differential distribution and varying significance as a driving cause of the pandemic on regional and global bases. A major challenge in designing and evaluating effective subtype-dependent candidate HIV-1 vaccines is the development of techniques for large-scale HIV genetic characterisation for documenting the true prevalence rates of HIV subtypes in developing countries. We have developed and validated an oligonucleotide probe hybridisation assay with high sensitivity and specificity to subtype HIV-1 CRF02_AG. To be effective against the full antigenic spectrum of primary HIV-1 isolates, candidate vaccines should contain immunogens of primary isolates representative for the whole HIV-1 antigenic spectrum. There is an urgent need to identify these immunogens and to improve their immunogenicity. The folding of gp41 into its fusogenic conformation, an obligate step in virus entry into the target cell, implies that the conformational properties of both the pre-hairpin as well as the trimer-of-hairpins structures may play a critical role in driving membrane fusion. We performed in collaboration with AlgoNomics N.V. in silico a full single amino acid substitution analysis, resulting in a Fold Compatible Database (FCD) for each conformation. Our results suggest that FCD predictions are in good agreement with the sequence variation as observed in well-validated e-gp41 sequences taken from our patient data or from a large public database. The FCD concept appears to be an efficient tool to restrict the number of substitutions that must be tested experimentally. It can be used to identify a limited set of substitutions to engineer pre-hairpin e-gp41 structural variants for use in drug screening programmes. In vaccine development, antibodies to such fusion intermediates would target conserved regions of the envelope proteins and therefore would be likely to neutralise a broad range of viral strains.
ITM promoter: G. van der Groen
ITM collaborators: W. Janssens, S. Coppens
Support: VIB (Flanders Inter-university Institute for Biotechnology)

The molecular biology team of the Microbiology department workingtowards a universal HIV vaccine.

Quantitative monitoring of neutralising antibodies in sera of HIV-vaccinated and/or HIV-infected individuals
Development and evaluation of a high throughput assay as a support for clinical anti-HIV vaccine trials. Quantification of neutralising antibodies currently requires handling infectious viruses, is labour intensive and expensive. We have developed a new in vitro neutralisation assay in which non-infectious HIV virus particles (pseudoviruses), which contain the envelope protein of representative HIV viruses, are used to infect well-standardised host cells harbouring the appropriate cellular receptors and a reporter gene. The latter allows us to quickly monitor HIV infection of the host cell. We have demonstrated comparable neutralisation data using either biological virus or pseudovirus. This assay has the added advantage that it can be performed in a Biohazard Level 2 facility, as compared to the classical neutralisation assay which needs to be done in Biohazard Level 3 facilities. As such, quantification of neutralising antibodies can be more easily integrated into laboratories of developing countries where vaccine trials are ongoing.
ITM promoter: G. van der Groen
ITM collaborators: L. Heyndrickx, H. Donners, D. Davis, K. Vereecken,T. -Vermoesen
External collaborators: Tibotec-Virco Ltd.
Support: IWT (Flemish Institute for the Promotion of Scientific-Technological Research in Industry)

Capacity strengthening of the National Reference Laboratory for HIV/STD, DR Congo
The main objective of this collaboration is to provide capacity strengthening of the HIV/AIDS reference lab (LNLS = Laboratoire Nationale de Lutte contre le Sida) in Kinshasa. This programme provides assistance to the LNLS in the diagnosis and follow-up of persons infected with HIV in DRC. The lab has proven to be able to maintain a relatively good level of quality, even under politically unstable circumstances. Currently, the perspectives of the laboratory are hopeful and the lab has obtained the official status of the HIV/STD reference lab for the DRC. The laboratory could be further equipped with the extra funding from the framework agreement. During this 5-year agreement, several members of the technical staff were trained at ITM. The extra funding provided by the AIDS Impulse Programme 2002 was used to computerise data collection, budget and stock management via the installation of computer networking, internet and e-mail. Several members of the technical staff were trained in our Immunology and Virology laboratory to update and improve quality control in the laboratory for the follow-up of HIV patients under anti-retroviral treatment in DRC.
ITM promoter: K. Fransen
ITM collaborators: L. Kestens, O. Courteille
External collaborators: J. Lepira, M. Tahiri (PNLS, D.R. Congo)



Unit of Immunology

Unit of Immunology: The more the merrier!

Endogenous mechanisms that protect HIV-exposed seronegative female sex workers against infection in Abidjan, Côte d’Ivoire:
In Abidjan, Côte d’Ivoire, 32% of female sex workers (FSWs) and 13% of their male clients are infected with HIV. HIV-negative female sex workers are frequently exposed to the virus based on reported numbers of clients per day, consistency in using condoms and duration of sex work, and are estimated to have on average 57 unprotected sexual exposures to HIV per year. FSWs with a duration of sex work of more than 3 years are unlikely to remain HIV-negative by chance alone, and possibly display an HIV-protective phenotype by one or several endogenous factors. Immunological factors associated with protection against HIV infection are studied in this population of HIV-exposed seronegative sex workers. Potential protective factors include both innate and adaptive immune responses. Expression levels of the HIV-1 coreceptor CXCR4, but not CCR5, were decreased among sex workers compared to controls. CXCR4 down-regulation was associated with a prolonged duration of sex work. However, an HIV-protective role for the decreased CXCR4 expression remains unclear, since HIV strains that are sexually transmitted predominantly use CCR5 as a coreceptor. Neither the CCR5 binding -chemokines nor Th1 and Th2 cytokines were found to differ among the sex workers, but they were found to have slightly increased T-cell activation levels compared to controls. Low-level HIV-specific T-helper responses were detected in a minority of sex workers. Levels of HIV-specific T cells were directly associated with the frequency of HIV exposure among the sex workers. Exposed sero-negative sex workers also had increased alloantigen-reactive humoral responses together with decreased cellular responses compared to controls.
ITM promoter: L. Kestens
ITM collaborators: W. Jennes, B. Vuylsteke
External collaborators: J. Nkengasong (Projet RETRO-CI, Abidjan, Côte d’Ivoire - Centers for Disease Control and Prevention, Atlanta, USA); L. Bélec (Retro-Ci Unité INSERM U430, Immunopathologie Humaine, Hôpital Broussais, Paris, France)
Support: Fund for Scientific Research - Flanders (FWO); Centers for Disease Control and Prevention, Atlanta, USA

Relative Viral Fitness in T-cells and Dendritic Cells in HIV Pathogenesis
Genetic variability is a hallmark of Human Immunodeficiency Viruses. Like any other RNA-virus, HIV exists as a quasi-species, i.e. a large set of distinct but related genomes. The error-prone nature of HIV’s reverse transcriptase enzyme is to blame for the rapid adaptation of the virus to immune pressure or antiretroviral treatment. Interest in viral fitness has increased in recent years. Fitness is a parameter that describes the replicative adaptation of a virus to its environment. Previously we demonstrated, together with our American colleagues, that relative viral fitness can be related to HIV pathogenesis. Now we are focusing on possible differences in viral fitness among HIV types, groups and subtypes. More specifically, we performed head-by-head dual infections/competitions with HIV-1 subtypes A, B, C, D and A/E, a selection of group O viruses and a variety of HIV-2 strains. The results are currently being analysed and written up in a paper. Furthermore, we are looking at the possible fitness-differences between syncytium-inducing and non syncytium-inducing viral clones and the fitness of circulating recombinant forms (more specifically CRF02_AG).
As the problem of drug resistance in HIV pathogenesis is reaching new heights, we have also planned to study the evolution of viral fitness in multidrug-resistant patients.
ITM promoter: G. Vanham
ITM collaborators: K. Ariën, L. Kestens, G. van der Groen, R. Colebunders, W. Janssens
External collaborators: E. J. Arts, M. E. Quinones-Mateu
Collaborating institutes: Case Western Reserve University, Cleveland, Ohio, USA; Lerner Research Institute, Cleveland Clinics, Cleveland, Ohio, USA
Support: Fund for Scientific Research - Flanders (FWO), Center For AIDS Research (CFAR-NIH) at Case Western Reserve University, Cleveland, Ohio, USA.

Development of a two-chamber system of epithelial cells and -dendritic – T cell co-cultures
We developed a co-culture system of dendritic cells (generated from monocytes with GM-CSF and IL-4) and autologous T cells to model early targets during sexual HIV transmission. This model seems well-suited for testing antiviral compounds like nucleoside and non-nucleoside reverse transcriptase inhibitors (N-RTI, or NN-RTI), acting between penetration of HIV into the target cell and integration into the host DNA. Other drugs, especially fusion and binding inhibitors, should act rather on the level of the epithelial cells of the vagina, cervix or rectum. To mimic this situation in our in vitro model, addition of epithelial cells to our present system is important. A two-chamber model will be evaluated, consisting of a lower chamber with dendritic cells and T cells and an upper chamber with epithelial cells. Preliminary experiments indicated that a confluent monolayer of endocervical or endometrial epithelial cells was a very potent barrier for HIV transmission, since no virus could be detected in the lower chamber even after 24 hours. This probably reflects the in vivo situation, in which the risk for sexual HIV transmission is rather low (less then 0.1%). However, in vivo, the vaginal epithelium is also often damaged through inflammation and infection with other pathogens, e.g. herpes simplex viruses, resulting in a higher transmission of HIV. To evaluate this situation, we will “damage” the in vitro epithelial layer by infecting it with a herpes simplex virus (HSV-2), followed by an infection with HIV. Transmission of HIV and subsequent infection of target cells (dendritic cells and T cells) will be evaluated, together with the impact of the dual infection on the viral replication in these target cells and the influence on their immune function. Once we have established this new model, several antiviral compounds can be evaluated for their potency in preventing or blocking infection.
ITM promoter: G. Vanham
ITM collaborators: Y. Van Herrewege, J. Michiels, P. Hertsens
External collaborators: R. Snoeck, G Andrei
Collaborating institutes: Rega Institute for Medical Research –Virology and Chemotherapy Lab
Support: CONRAD Organisation

Evaluation of antiviral drugs as microbicides for prevention of sexual HIV transmission
There is an urgent need for HIV prevention, mainly in resource-poor settings. Condoms are not always acceptable, and therefore vaginal microbicidal drugs may be an acceptable alternative.
We used an in vitro model, based on monocyte (MO)-derived dendritic cells (DC) and CD4 T cells, to evaluate the potency of several antiviral drugs as microbicides. The MO-DC constitute a model of mucosal DC, early targets during sexual transmission, playing a crucial role in the transfer of HIV to T cells, but also in the induction of a beneficial immune response against HIV.
In preliminary studies, we showed that polyanionic binding-inhibitors, fusion inhibitors (T-20) and co-receptor blockers could suppress viral growth at concentrations of 10-100 µg/ml. However, these products were unable to really prevent HIV infection, unless they were used in concentrations that produced immune-suppression. By contrast, the NN-RTI UC-781 and TMC-120 apparently could prevent infection at nanomolar concentrations, whereas immune suppression was only present in the micromolar range. Therefore, we further explored the use of several other, new NN-RTIs, discovered at the Centre for Molecular Design of Janssen Pharmaceutics. The results of these experiments were very promising and showed that these new drugs were amongst the most potent ones we have tested so far. Complete suppression of viral replication was possible at concentrations of 10-100 nanomolar, whereas immune suppression only occurs at concentrations of 1-10 micromolar. A selection of these compounds was evaluated for their potency to prevent infection if treatment was limited to 24 hours. In these experiments, prevention of infection was possible at drug concentrations of 10-100 nanomolar. These results indicate that a short treatment, at the time of incubation of target cells with virus, can prevent infection of these target cells.
ITM promoter: G. Vanham
ITM collaborators: Y. Van Herrewege, H. Njai, J. Michiels
External collaborators: P. Lewi
Collaborating institutes: Centre for Molecular Design – Janssen Pharmaceutics
Support: Centre for Molecular Design – Janssen Pharmaceutics.

Biological follow-up of HIV/AIDS patients in Senegal:

The general aims of this project are to strengthen the capacity of the Virology and Bacteriology laboratory of the Faculty of Medicine and Pharmacy at the University “Cheikh Anta Diop“ in Dakar, to train scientific and technical personnel, and to evaluate new and mobile methods for the immunological follow-up of HIV/AIDS patients. The introduction of antiretroviral treatment for HIV in Africa requires affordable virological and immunological follow-up. Today such follow-up is difficult, particularly in remote reference centres. Simplified techniques are not always reliable and are often labour intensive. Sophisticated methods such as flow cytometry to perform CD4 counts are too expensive and require expensive reagents. In this project we evaluated a new, affordable and mobile cytometer to measure CD4 counts. The results of the evaluation are encouraging and show that follow-up can indeed be affordable, although well-trained personnel are required to ensure good quality.
ITM promoter: L. Kestens
ITM collaborators: P. Ondoa, C. Vereecken
External collaborators: S. Mboup, T. Ndeye (Laboratoire de Virologie & Bactériologie, Faculté de Médecine et Pharmacie de l’Université Cheikh Anta Diop, Dakar and Centre Hospitalier Universitaire (CHU))
Support: DGDC: AIDS Impulse Programme 2002.

Product development for and with developing countries
The main objectives of this project are to identify and optimise laboratory tests to demonstrate protective immune responses (virus neutralisation) and cellular immune responses; to adapt these methods for use in developing countries; and to train personnel from developing countries for laboratory monitoring after antiretroviral therapy.
ITM has done a great deal of research on the diversity of HIV in developing countries which has given us knowledge which is essential for vaccine development. Current laboratory tests are unsuitable or insufficiently adapted to evaluate the protective capacity of anti-HIV vaccines under field conditions.
With the financial support of this project we were able to cover the salary cost of the scientific staff of the department, renew the “state-of-the-art” equipment of the laboratories, and further develop laboratory tests to measure virus neutralisation and cellular immune responses. With regard to laboratory testing, special attention was also paid to the evaluation of a screening test to monitor recovery of the immune system after antiretroviral therapy. In that regard, the HIV p24 antigen test has been assessed as a surrogate marker of HIV mRNA viral. Alternative tests for CD4 counting were evaluated. Laboratory training was provided to 6 technicians and 2 scientists from D.R. Congo, Côte d’Ivoire, Senegal and Rwanda.
ITM promoters: L. Kestens, G. van der Groen
Support: DGDC - AIDS Impulse Programme

Clinical management of HIV/AIDS in Peru
Within the existing Institutional collaboration between the Institutes of Tropical Medicine of Lima (IMTAVH) and ITM Antwerp, the following additional objectives were pursued through the Aids Impulse Programme: acceleration and upscaling of clinical studies to improve the management and treatment of AIDS patients in Lima and Peru; linking these activities to prevention of HIV transmission in Lima, with adolescents and pregnant women as special target groups, and to social action; and establishment of an immunological laboratory at IMTVAH for the diagnostic follow-up of HIV infection, AIDS treatment, and related research on HIV and infectious diseases. The laboratory was fully equipped in 2002 (FACSCalibur, incubators, laminar flow) and Guido Vanham spent two months (October-November 2002) at the Institute to train scientific personnel in laboratory immunology and immunological follow-up of HIV/AIDS patients under antiretroviral treatment.
ITM promoter: G. Vanham
ITM collaborators: T. Verdonck
External collaborators: E. Gotuzzo (IMTAVH, Lima, Peru)
Support: DGDC - AIDS Impulse Programme


Unit of Mycobacteriology

TUBERCULOSIS (TB)

Tuberculosis: Drug resistance surveillance.
The general aim of this project is to document the prevalence of drug-resistant TB in various continents and regions, both to provide useful information for treatment regimens and to evaluate TB control programmes. Our studies made clear that two approaches could be applied for this purpose: representative sample surveys that can be repeated every 5 years, and follow-up of drug resistance in relapse and failure cases. The focus lies on first-line drugs, but second-line drugs are also included if possible and appropriate in the settings. Furthermore, this project aims to evaluate various factors influencing the outcome of treatment regimens, such as the use of intermittent DOT regimens versus non-DOT daily regimens in the Philippines, and to evaluate different MDRTB regimens in Bangladesh. Besides resistance surveillance, operational research is included to improve direct microscopy, optimise specimen transport for culture, and optimise cheap, fast and easy alternatives to the classical drug-susceptibility tests.
ITM promoter: F. Portaels
ITM collaborators: A. Van Deun, L. Rigouts, C. Shamputa
External collaborators: DF TB Control Projects from Bangladesh, India and Congo-Kinshasa; National TB Programmes of Burundi, Rwanda and the Philippines; ICRC Geneva; MSF Belgium
Support: Damien Foundation, Belgium

Tuberculosis: Investigation of the development of resistance
The general aim of this project is to gain a better understanding of the creation and amplification of resistance to first-line drugs by TB bacilli in the course of treatment. Secondly, we want to estimate to what extent reinfection and initial multiple infection with various M. tuberculosis strains play a role in cases of apparent failure or relapse after treatment. Important types of resistance are always man-made, due to therapeutic errors, and especially frequent where serious economic and management problems occur. Original short-course treatment regimens were based on daily dosing, and rifampicin was used exclusively in a quadruple combination in the intensive phase. The replacement of daily by intermittent, twice or thrice weekly dosing, and the use of R in combination with Isoniazide in the second phase, are two recent trends meant to make directly observed therapy (DOT) more feasible. In strictly-controlled trials this yields results which are just as good as daily dosing, but it is far from clear to what extent these regimens may also cause resistance under field conditions. To achieve these objectives, drug-susceptibility testing (including estimation of minimal inhibitory concentrations) and DNA-fingerprinting techniques will be applied on serial isolates from patients under various treatment regimens, as well as on duplicate pre-treatment isolates.
ITM promoter: F. Portaels
ITM collaborators: A. Van Deun, L. Rigouts, C. Shamputa
External collaborators: DF TB Control Projects from Bangladesh, India and Congo-Kinshasa; National TB Programmes of Burundi, Rwanda and the Philippines
Support: Damien Foundation, Belgium

Tuberculosis: Search for mixed infections in patients from the penitentiary hospital in Tbilissi, Georgia
As the prevalence of MDRTB was found to be high (9% of isolates) in previous investigations, it was decided to treat all patients in the hospital with the standardised re-treatment regimen (Category 2) from the WHO, regardless of the patients’ treatment histories. The three main objectives of this study are, firstly, to determine the rate of re-infection with an MDRTB strain in this high-risk population. Secondly, to determine the risk of the ‘amplifier effect’, i.e. the creation of additional resistance during a standardised treatment in this setting with a high level of initial resistance. And thirdly, to determine the rate of mixed infections, i.e. the simultaneous infection with different M. tuberculosis isolates, and the extent to which treatment outcome is influenced by this phenomenon. Therefore, a cohort follow-up study including at least 300 smear-positive TB patients will be organised. Preliminary results have shown the presence of possible mixed infections in about 8% of pulmonary TB patients in this setting. Furthermore, this study will make it possible to determine the prevalence of resistance to second-line drugs in this setting with a relatively high MDRTB rate.
ITM promoter: F. Portaels
ITM collaborators: L. Rigouts, C. Shamputa
External collaborators: ICRC Geneva
Support: European Community

Joint research project on tuberculosis control in The Free State, South Africa: from infection to cure
Most importantly, the envisaged study will take the researchers deep into the unknown world of TB patients, and take up the daunting challenge of achieving cure through a protracted treatment schedule closely impacted by health service policy and organisation, and within a context of scarce resources. The study is to follow the broad path of the TB ‘career’ from diagnosis through the intensive and follow-up phases up to the variable outcomes of a treatment process in South Africa provided almost exclusively by the government-run primary health care (PHC) services. The study is to be comprehensive in its scope, covering - in addition to the social and psychological experience - patterns of adherence to treatment among patients, costs to TB patients and their households, and a diverse range of health system factors. Parallel microbiological analyses will document the findings. All these will be investigated in the light of the South African government’s policy and programme to strengthen PHC and devolve resources and decision-making to district and local levels. Although the research has not run its full course, some weak points and strengths of the local TB programmes could already be identified, leading to the first practical interventions. The decision was taken to (re-)train all clinic TB coordinators in the National Tuberculosis Control Program, and to (re-)train 90% of all staff at the clinics. Furthermore, stocktaking of the positive elements in TB control in this province of South Africa provided useful information for other provinces.
Promoter: H. Meulemans (Political and Social Sciences, UIA, Antwerp, Belgium)
ITM collaborators: F. Portaels, L. Rigouts
External collaborators: J. Vanhoutte, D. De Graeve, L. Pauwels,C. Timmerman (University of Antwerp, Belgium, Departments of Political and Social Sciences and Economics); D. van Rensburg, C. Heunis, E. Janse van Rensburg, Z. Matebesi, Center for Health Systems Research &Development); R. Chapman, A. Vanderspoel (Department of Health, University of Free State, Bloemfontein, South Africa)
Support: Bilateral collaboration funded by the Flemish Government.

Concerted Action Project: Improved diagnosis, drug resistance detection and control of tuberculosis in Latin America
Tuberculosis (TB), which had been considered a disease of the past, has re-emerged as a serious problem in Europe as well as in the developing world. Even more alarming is the appearance of drug resistance. There are foci in Eastern Europe where one in four cases are caused by multidrug-resistant TB (MDR-TB) strains. A strategic obstacle for improving TB control is the lack of easy and inexpensive techniques that can replace the slow and laborious conventional methods for diagnosing TB and detecting drug resistance. However, there are newer, more appropriate methodologies that warrant further evaluation. This Concerted Action will develop, adapt and evaluate some of these promising new tools. It will build upon collaborations that have developed between a network of European researchers and their Latin American colleagues. Preliminary studies by this group have begun to address some of the objectives in this project, and the work outlined will expand these collaborations to develop useful and appropriate methodologies and test them in prospective clinical trials in high TB prevalence areas. It will also gather information on the epidemiology of MDR-TB in Latin America.
The first general meeting held in Antwerp (July 2002) to launch the project was a great success.
ITM promoter: F. Portaels
ITM collaborators: J.C. Palomino, A. Martin, L. Rigouts
External collaborators: A. Bernardelli (SENASA); M. Camacho (Instituto Nacional de Laboratorios de Salud); A. Cataldi (Institute of Biotechnology CICVyA-INTA); L. Chacon (Ministerio de Salud de Nicaragua); A.P. da Silva (Universidade do Rio Grande); M.A. da Silva Telles (Instituto Adolfo Lutz); P. del Portillo (Corporacion CORPOGEN); J. de Waard (Instituto de Biomedicina); I. Estrada (Escuela Nacional de Ciencias Biologicas); C. Espitia (Inst. Investigaciones Biomédicas UNAM); J. Fernandez, M. Velasco (Inst. de Salud Publica de Chile); M.J. Garcia (Universidad Autonoma de Madrid); B. Gicquel (Institut Pasteur); G. Gonzalez y Merchand, J. Luna-Herrera (Instituto Politécnico Nacional); M.I. Guerrero, C. Leon (Instituto Nacional de Salud); P.W.M. Hermans (Erasmus University Rotterdam); R. Hernandez-Pando(Salvador Zubiran); A. Kritski (Universidad Federal de Rio de Janeiro);S.C. Leão (Universidade Federal de São Paulo) ; J.L. Maestre Mesa,E. Montoro (Instituto de Medicina Tropical Petro Kouri); C. Martin, I. Otal,S. Samper (Universidad de Zaragoza); J. Robledo, G.I. Mejia (Corporacion para Investigaciones Biologicas); F. Mello (Federal University of Rio de Janeiro – UFRJ); N. Morcillo (Hospital Centrangolo); V. Ritacco (National Institute for Infectious Diseases); M. Romano (Institute of Biotechnology INTA); E. Roxo (Instituto Biologico); L. Salazar (Inst. Venezolano de Invest. Cientificas IVIC); P. Suffys (Fundacion Oswaldo Cruz); H. Takiff (Inst. Venezolano de Invest. Cientificas IVIC); D. van Soolingen (National Institute of Public Health and the Environment); J. Zuria (Laboratorio Clinico)
Support: INCO Programme of the European Commission

BURULI ULCER

Development of new methods for identifying the reservoir of Mycobacterium ulcerans and for better understanding the mode of infection of Buruli ulcer.
One approach to improving our understanding of the ecology of Mycobacterium ulcerans has been to study the molecular epidemiology of M. ulcerans, for which several methods have been used: IS2404-RFLP, analysis of 16S rRNA, AFLP and DR analysis (spoligotyping). The discriminatory power of these fingerprinting analyses was limited, since isolates from different continents showed different patterns, but it was not possible to differentiate isolates from the same country.
However, fingerprinting methods have been successfully used to clarify the phylogenetic position of M. ulcerans in relation to its closely-related species e.g. M. marinum. This study has been completed and found the presence of intermediary forms between M. marinum and M. ulcerans, which confirms the hypothesis of Stinear et al. that M. ulcerans acts as a progenitor of M. marinum by acquiring DNA elements from the environment.
ITM promoter: F. Portaels
ITM collaborators: A. Ablordey, K. Chemlal
Support: Damien Foundation, Belgium, Fund for Scientific Research - Flanders

Buruli ulcer in Benin
The general aim of this project is to better control Buruli ulcer (BU) in Benin by using a multidisciplinary approach. Trends in BU patients, as seen in a rural hospital of southern Benin, were evaluated on 1700 cases of Mycobacterium ulcerans disease cases (Buruli ulcer) treated at the “Centre Sanitaire et Nutritionnel Gbemonten” in Zagnanado (Benin). The patients lived in the 4 regions of Southern Benin: Atlantique, Mono, Oueme and Zou, with the largest number coming from the Zou Region, where the centre is located. In this region, the 1999 detection rate of Buruli ulcer (21.5/100 000 pop.) exceeds those of leprosy (13.4/100 000 pop.) and tuberculosis (20.2/100 000 pop.).
The median age of patients is 15 years (q1=7, q3=30). There is a significant difference in the age and sex composition, with more males in the under-20 year-old patients, while females are more frequent in those over 20 years old.
More than 13% of the patients have osteomyelitis. The form of the disease is related to the period of delay in presentation to the hospital. The median hospitalisation time was reduced from 267 days in 1989 to 32 days in 2002. It was shown that effective BCG vaccination at birth protected BU patients against developing severe forms of this disease such as osteomyelitis.
The influence of HIV infection on BU was also studied. It was confirmed that HIV infection plays an important role in the development and extent of dissemination of BU.
A multidisciplinary approach (e.g. education, training and treatment) is required for optimal management of M. ulcerans disease to decrease morbidity and socio-economic impact of Buruli ulcer on rural populations in Benin.
ITM promoter: F. Portaels
ITM collaborators: M. Debacker, C. Zinsou
External collaborators: A. Guédénon (PNLUB, Benin)
Support: DGDC - AIDS Impulse Programme, Damien Foundation, Belgium

NON-TUBERCULOSIS MYCOBACTERIA

Development of diagnostic tools and medical prophylaxis to control bovine paratuberculosis
This collaboration seeks to improve direct and indirect diagnostic tests for bovine paratuberculosis, a disease caused by Mycobacterium avium subsp paratuberculosis. Direct detection of these bacilli in stool will be emphasised by the development of an immunocapture assay to detect specific antigens and the optimisation of a PCR assay. Improvement of indirect diagnostic tests to estimate the TH2 and TH1 cellular responses by quantifying IL-5 and IL-10 or GM-CSF, respectively, will make it possible to determine the type of response and to see whether they can help in predicting the stage of infection. Furthermore, the immunological capacity and efficiency of plasmid vaccines based on genes encoding for antigens A85A and A85B will be evaluated in existing mouse models and newly developed models in small ruminants. Finally, a newly developed DNA-fingerprinting method based on the insertion sequence IS900 (Flanking Sequence Polymorphism, FSP) will be optimised and evaluated on a panel of well-documented M. paratuberculosis isolates originating from wild and captive animals.
Promoter: J. Godfroid, Centre d’Etude et de Recherches Vétérinaires et Agrochimiques (CERVA), Belgium
ITM collaborators: F. Portaels, L. Rigouts
External collaborators: J. Godfroid, K. Walravens (CERVA); K. Huygens (Pasteur Institute Brussels, Belgium)
Support: Flemish Government.


Unit of STD/HIV Research and Intervention

STD Unit: together we test for gonorrhoea and much, much more…


Study on adolescents’ sexual behaviour and on contacts with sex workers in four African cities with different levels of HIV infection.
This project is a follow-up to The Multicentre Study on Factors determining the Differential Spread of HIV in four African Cities, which had left a number of questions unanswered. The aim of this project is to gain better insights into the sexual behaviour of young people and into networking between commercial sex workers and their clients in four cities in sub-Saharan Africa. While the multicentre study employed a standardised questionnaire with closed questions, this study employed qualitative methods. In each town, in-depth interviews on adolescent sexuality were conducted with 75 young men and 75 young women (aged 15 to 20) using purposive quota sampling, with age, educational and socio-economic variables. These interviews were transcribed verbatim and processed for analysis with qualitative analysis software (Atlas.ti). Open-ended questions dealt with current and past sexual experiences and contextual risk factors. At the same time, interviews were also done with 75 men (aged 20 to 49) in the general population and 50-100 clients of sex workers; though data from these interviews do not focus directly on adolescent behaviour, they may contain important information on men’s contacts with young adolescents or their own experiences as adolescents. 8 focus group discussions on perceptions of adolescent sexuality were also conducted in each town, as well as formal and informal observations of situations related to adolescent sexuality (such as youth attendance in video halls, dance parties, funerals, bars, etc.).
Data analysis is nearly completed for two sites (Kisumu in Kenya and Ndola in Zambia) and is ongoing for the other two sites (Cotonou in Benin and Yaoundé in Cameroon). In December 2002, results of the study in Kisumu were presented to stakeholders, including the provincial medical authorities, NGOs and the young people themselves through a number of workshops.
ITM promoter: A. Buvé
ITM collaborators: P. Remes
External collaborators: Provincial Medical Office, Kisumu (Nyanza Province, Kenya); Tropical Diseases Research Centre (Ndola, Zambia); Centre de Recherche en Reproduction Humaine et en Démographie (Cotonou, Benin); Université Catholique d’Afrique Centrale (Yaoundé, Cameroon)
Support: The Wellcome Trust.

Impact of a comprehensive sexual and reproductive health programme for adolescents in Kenya
In 2001 the Unit of STD/HIV Research and Intervention conducted a youth needs assessment in Nyanza Province, Western Kenya, which is the region in Kenya which has been hardest hit by the HIV/AIDS epidemic (HIV infection prevalence is 25% among adults in the capital Kisumu). This assessment highlighted the inadequate responses of government and community to young people’s most pressing needs. A multi-component programme was designed based on the proposals that the young themselves brought up and on current knowledge of what has proven successful. It aims to provide appropriate sex and reproductive health education, training in life skills, personal counselling and customised health services to young people aged 10 to 20 years in two districts in Nyanza Province. It will also offer opportunities in vocational training and livelihood skills, and support income-generating activities. A network of youth peer educators will conduct outreach activities in the communities and refer young people to the youth centre and a youth-friendly clinic. The main objective of the youth programme is to reduce the spread of HIV infection by reducing unsafe sexual behaviour. The effects of the programme will be assessed by a comparison between a baseline survey among youth in the general population and a follow-up survey.
The project started in August 2002 and the first youth centre is expected to open in March 2003. The baseline survey is planned for March-April 2003.
ITM promoter: A. Buvé
ITM collaborators: H. Vandenhoudt
External collaborators: Centers for Disease Control and Prevention (Atlanta, USA); Kenya Medical Research Institute (Kisumu, Kenya); Provincial Medical Office, Kisumu (Nyanza Province, Kenya).
Support: Centers for Disease Control and Prevention

Opportunities and vulnerabilities in the lives of very young adolescents.
This project consists of three parts:
1. A literature review is being conducted to explore published studies relating to very young adolescents. The review will focus on interrelated issues and domains surrounding first sexual experience. This includes: age and circumstances of first sex, relationship characteristics, sexual networking patterns and partner selection, age differences between sexual partners, use of health services, substance use and other contextual factors that either strengthen protective behaviours or enhance risky behaviours.
2. Through the “Study on Adolescents’ sexual behaviour and on contacts with sex workers in four African cities with different levels of HIV infection” qualitative research was conducted on sexual interactions of groups at high risk of HIV infection in four urban populations in Africa to reach a better understanding of their role in the dynamics of HIV infection and to identify prevention strategies (see above). The qualitative interviews with 150 young people aged 15-19 in each of the 4 cities are analysed to retrospectively check sexuality-related experiences which occurred when these youth were 10-to-14 years old, and to describe contextual factors that strengthen protective behaviour or enhance risky behaviours during this period and later in adolescence.
3. During site visits to Kisumu and Ndola, results from the qualitative study of youth sexuality are discussed with stakeholders and community members. During the site visits, an assessment will also be made of interventions that are already being carried out to address the specific conditions of 10-to-14 year-olds and/or explore which kinds of interventions local organisations/institutions can implement with and among very young adolescents in order to delay first sex, reduce sexual partners, promote faithfulness and monogamous relationships, and promote condom use.
ITM promoter: A. Buvé
ITM collaborators: P. Remes, J. Van Wingerden
External collaborators: Provincial Medical Office, Kisumu (Nyanza -Province, Kenya); Tropical Diseases Research Centre (Ndola, Zambia).
Support: UNAIDS.

Study on the utilisation of antiretroviral therapy in Kinshasa (Democratic Republic of Congo) and Pointe-Noire (Congo-Brazzaville) and financial feasibility study of an ambulatory clinicfor HIV-infected patients
The objective of this study was to assess the utilisation of antiretroviral drugs (ART) in the private sector in Kinshasa, the drugs not yet being available in the public sector or in NGO projects. Interviews were held with physicians, patients coming to the reference laboratory of the Programme National de Lutte contre le SIDA for a CD4 count and clients of NGOs that care for HIV-infected patients. Altogether, the physicians who were interviewed had more than 300 patients on ART. Most prescriptions were adequate, but there was a major problem for patients to access the drugs. Twenty-eight samples of patients from Kinshasa on ART were tested for resistance. Testing of samples from patients in Pointe-Noire is still ongoing.
Based on data from the Centre de Traitement Ambulatoire at Pointe-Noire, which is supported by the French Red Cross, and a spreadsheet model, estimates were made – under varying assumptions – of the costs of caring for HIV-infected patients at the CTA in Kinshasa which is being set up by Médecins sans Frontières Belgium.
Preliminary data from these studies as well as the spreadsheet model have been presented in Kinshasa at a workshop on care for HIV-infected patients.
ITM promoter: A. Buvé
ITM collaborators: B. Ostyn , R. Colebunders, K. Fransen
External institutes/collaborators: Programme National de Lutte contre le SIDA (Kinshasa, DRC); GTZ (Kinshasa, DRC); MSF Belgium
Support: DGDC - AIDS Impulse Programme.

Study on the prevalence of HIV infection among women attending the clinic of Matonge and study on the aetiology of STDs in women and men attending the clinic of Matonge and the Centre Victoire in Kinshasa (Democratic Republic of Congo)
In the 1980s and up to 1991, the Department of Microbiology was a partner in Projet SIDA. One of the responsibilities of the department was to run a dedicated clinic for sex workers in Matonge, the “hot area” of Kinshasa. Following the political turmoil of 1991, the running of the clinic of Matonge was taken over by MSF. This organisation also opened an STD clinic on the Place de la Victoire in the same neighbourhood. The clinic of Matonge is attended by sex workers, while the Centre Victoire is open to men and women with STI-related complaints. The objectives of the present study was to assess the prevalence of HIV infection in sex workers, 10 years after the last study had been conducted, and to measure the prevalence of other STIs in sex workers and in men and women with STD-related complaints. Preliminary data suggest that the prevalence of HIV infection among sex workers remains relatively low in Kinshasa.
ITM promoter: A. Buvé
ITM collaborators: B. Ostyn, R. Colebunders, K. Fransen
External collaborators: Programme National de Lutte contre le SIDA (Kinshasa, DRC); GTZ (Kinshasa, DRC); MSF Belgium
Support: DGDC - AIDS Impulse Programme

Operational research in STI and related services for women in high-risk situations in Cambodia and Thailand (The ORISS Project Cambodia-Thailand)
The European Commission at the end of 2002 approved this project. Project activities will begin in February 2003. The Project aims to provide the National Health Authorities in Cambodia and Thailand with a better understanding of high-risk situations among vulnerable women, and of reproductive health needs, in order to help them develop appropriate policies and strategies.
The project consists of three inter-related studies:
1. Validation of the management protocol of vaginal discharge in Cambodia.
2. Assessment of circumstances of sex work and mobility among female sex workers in Cambodia.
3. Assessment of circumstances of mobility and of reproductive health needs among vulnerable women in Thailand.
The latter two studies will employ qualitative research methods as well as classical epidemiological methods.
ITM promoter: A. Buvé
ITM collaborators: F. Crabbé
External collaborators: R. Coutinho (Municipal Health Service, Amsterdam, The Netherlands); S. Hean (The Centre for Advanced Study, Phnom Penh, Cambodia); B. Pattanaik (Alliance against Traffic in Women Foundation, Bangkok, Thailand)
Support: European Commission.

Care and Prevention of Sexually Transmitted Diseases in Cambodia
In early 1997 the Unit was contracted by the European Commission to implement a 4-year project, “Care and Prevention of Sexually Transmitted Diseases in Cambodia”, in close collaboration with the National Centre for HIV/AIDS, Dermatology and STD (the National HIV/AIDS Control Programme in Cambodia). This project ended in January 2001 and activities that had started under the EC project were maintained in Phnom Penh and in Sihanoukville, thanks to funding from the Institute of Tropical Medicine through an agreement between the Institute and the Ministry of Cooperation and Development. The project consists of the provision of technical assistance in matters of STD control, logistical support and capacity building, as well as support with supplies. In 2001 the project benefited from extra financial inputs through the AIDS Impulse Programme.
ITM promoters: M. Laga, A. Buvé
ITM collaborators: F. Crabbé
External collaborators: C.V. Mean (National Centre for HIV/AIDS, Dermatology and STD, Phnom Penh, Cambodia)
Support: DGDC

Institutional strengthening and collaboration with the Institut National de Santé Publique (Abidjan, Côte d’Ivoire)
Since 1998 the Unit has a collaboration with the Institut National de Santé Publique (INSP) in Abidjan, Côte d’Ivoire. This collaboration is funded by DGDC and in 2001 an additional budget was secured within the AIDS Impulse Programme. This additional funding permitted further strengthening of the infrastructure at the INSP (library and computer network). In addition, the collaboration consists of training staff of the INSP (in computer skills and project management), the organisation of regional courses in epidemiology (in collaboration with Projet Retro-CI of the Centers for Disease Control and Prevention) and joint research projects. The research activities which the Unit carries out in collaboration with the INSP and Projet Retro-CI are concentrated around the Clinique de Confiance, a dedicated clinic for sex workers in Abidjan. The clinic conducts HIV surveillance among sex workers and in 2002 initiated a study on the acceptability of diaphragms by sex workers.
However, research activities had to be temporarily suspended in the last quarter of 2002 because of political instability.
ITM promoters: M. Laga / A. Buvé
ITM collaborators: B. Vuylsteke (STD/HIV Research and Intervention Unit)
External collaborators: J. Diarra (INSP, Abidjan, Côte d’Ivoire); Projet Retro-CI, Abidjan
Support: DGDC

Prevention and care of STIs and HIV infection among African migrants in Flanders
The activities of this project used to be concentrated on Antwerp and Ghent. However, in 2002 an agreement was made with the Province of Flemish-Brabant to initiate activities in Leuven as well. The objectives of the programme are to improve knowledge about STIs and HIV and to create networks that can provide culturally-adapted STI/HIV prevention and care services. These networks also enable contact with hard-to-reach groups of migrants and ensure the sustainability of the activities. Prevention activities include meetings with African migrants and training of peer educators. The self-help group Muungano targets HIV-infected migrants. The members of the group meet on a monthly basis to discuss subjects such as therapy adherence and to strengthen solidarity within the group.
On World AIDS Day (December 1st) a meeting was organised where religious leaders of different denominations from the African community were invited to talk about AIDS. The opening speech of the meeting was held by Mrs M Vogels, Flemish Minister of Health and Social Affairs. The meeting was very well attended and drew our attention to another potentially important stakeholder in HIV prevention: religious leaders (see Focus on). In 2003 we will explore the possibility of having more frequent meetings with these community leaders and to involve them more closely in HIV prevention in Belgium, but also in their countries of origin.
ITM promoters: M. Laga, A. Buvé
ITM collaborators: R. Struelens, L. Manirankunde, T. Alou, I. Kint
External collaborators: African community, Belgium
Support: Flemish Government, Provinces of East-Flanders, Antwerp and Flemish-Brabant, European Commission (AIDS and mobility), DGDC

Policy Research on integration of reproductive health services in primary health care services and on integrated AIDS policy
Thérèse Delvaux is responsible for policy research on integration of reproductive health services in primary health care services. She spent most of her time on the topic of new developments in the area of prevention of mother-to-child transmission of HIV. In collaboration with Carine Ronsmans of the London School of Hygiene and Tropical Medicine, the INSP in Abidjan and Projet Retro-CI, she worked on a study of the effects of introducing a programme of mother-to-child transmission prevention on the quality of obstetric services in San Pedro, a coastal town in Côte d’Ivoire. The baseline assessment has been done but the introduction of the intervention is now delayed because of the political problems in Côte d’Ivoire.
Hedwig Maex, who has extensive experience with DGDC and VVOB, was contracted to write a document on the mainstreaming of HIV/AIDS in Belgian co-operation programmes. The document has been discussed with the staff of DGDC. Hedwig went to present the need for an integrated approach to AIDS and for mainstreaming to a meeting of attachés in -Ouagadougou (Burkina Faso), where her presentation was very favourably received.
ITM promoters: M. Laga, A. Buvé
ITM collaborators: T. Delvaux, H. Maex
External collaborators: J. Diarra (INSP, Abidjan, Côte d’Ivoire)
Support: DGDC

Technical assistance to USAID-funded projects in developing countries
Since May 1998 the Unit has been subcontracted by FHI (Family Health International) to provide technical assistance to STI control projects in several African countries. A new agreement was signed in 2002. FHI has asked the Unit to extend the scope of its work. Technical assistance will be provided in the areas of HIV care with antiretrovirals and STI control. Two staff members from the Unit are working part-time for FHI. Bart Ostyn has done a first evaluative mission to Rwanda, where FHI intends to start an ART-access programme.
François Crabbé, who is based in Cambodia, has also been asked by FHI to provide technical assistance for a survey on sexually-transmitted infections in East Timor. He made a first visit to East Timor and has written a protocol for the survey.
ITM promoters: M. Laga, A. Buvé
ITM collaborators: B. Ostyn, J. Vandepitte, R. Colebunders
External collaborators: Family Health International.
Support: Family Health International

Research on microbicides
In the past year, the STI laboratory of the Unit has provided lab support (training, quality control, evaluation) to several projects on microbicides, including a WHO study on cellulose sulphate in Uganda, India and Nigeria and a CONRAD study in India. The Unit has made an agreement with CONRAD, a US-based NGO, for further collaboration. The Unit will provide technical assistance to STI laboratories in developing countries and will conduct phase 1 trials in Antwerp.
The Unit of Immunology and the Unit of STD/HIV Research and Intervention are also collaborating in a European consortium that will submit a major grant application to the 6th Framework Programme of the European Commission.
In May 2002 the Unit organised and hosted the Microbicides 2002 Conference. Marie Laga was the conference chair and Lut Van Damme the scientific coordinator (see also Focus on).
ITM promoters: M. Laga, A. Buvé
ITM collaborators: V. Jespers, G. Vanham
External collaborators: WHO; L. Van Damme, M. Callahan CONRAD
Support: WHO, CONRAD

Epidemiology of Trichomonas vaginalis: prevalence of Trichomonas sp., and sexual and hygienic risk factors associated with genital trichomoniasis
Trichomonas vaginalis (Tv) is highly prevalent among women in Africa. Tv is believed to be sexually transmitted. However, the Multicentre study on factors determining the differential spread of HIV in four African Cities showed a high prevalence of Tv infection among girls in Ndola (Zambia) who denied ever having had sex. Data from a study in Mwanza on young girls also indicates a high prevalence of Tv, while the prevalence in men is relatively low. This led us to postulate that not all of these vaginal infections in young girls were due to Tv, but that at least a certain proportion of them are caused by related species such as P. hominis and/or T. tenax, which colonise the vagina from the digestive tract. These parasites do grow on conventional media (own data, unpublished) and cannot be distinguished from Tv by direct microscopy. The objectives of this project are to develop PCR techniques for differentiating different species of trichomonads and to use these techniques to study the epidemiology of the different species and risk factors for acquiring the parasite.
There is no funding for this project yet, and in 2002 the work concentrated on developing the PCR techniques.
ITM promoters: A. Buvé, E. Van Dyck
ITM collaborators: T. Crucitti
External collaborators: R. Musonda (Tropical Diseases Research Centre, Ndola, Zambia); Y. Adu-Sarkodie (School of Medical Sciences, Kwame Nkrumah University of Science and Technology, Kumasi, Ghana); D. Mabey (London School of Hygiene and Tropical Medicine)
Support: Pending


Kengo, Kenya. Renovation of an old building that will serve as a youth centre.


Publications in international peer-reviewed journals

Balzarini J, Van Herrewege Y, Vanham G. Metabolic activation of nucleoside and nucleotide reverse transcriptase inhibitors in dendritic and Langerhans cells. AIDS 2002; 16: 2159-2163.
Beelaert G, Vercauteren G, Fransen K, Mangelschots M, De Rooy M, Garcia-Ribas S, van der Groen G. Comparative evaluation of eight commercial enzyme linked immuno-sorbent assays and 14 simple assays for detection of antibodies to HIV. J Virol Methods 2002; 105: 197-206.
Bortolotti V, Buvé A. Prophylaxis of opportunistic infections in HIV-infected adults in sub-Saharan Africa: opportunities and obstacles. AIDS 2002; 16: 1309-1317.
Boutonnet N, Janssens W, Boutton C, Verschelde JL, Heyndrickx L, Beirnaert E, van der Groen G, Lasters I. Comparison of predicted scaffold-compatible sequence variation in the triple-hairpin structure of human immunodeficiency virus type 1 gp41 with patient data. J Virol 2002; 76: 7595-7606.
Buvé A. AIDS in Africa [reply]. Lancet 2002; 360: 1423-1424.
Buvé A. HIV epidemics in Africa: what explains the variations in HIV prevalence. IUBMB Life 2002; 53: 193-195.
Buvé A, Bishikwabo-Nsarhaza K, Mutangadura G. The spread and effect of HIV-1 infection in sub-Saharan Africa. Lancet 2002; 359: 2011-2017.
Camps K, Jorens PG, Demey HE, Pattyn SR, Ieven M. Clinical significance of herpes simplex virus in the lower respiratory tract of critically ill patients. Eur J Clin Microbiol Infect Dis 2002; 21: 758-759.
Chemlal K, Huys G, Laval F, Vincent V, Savage C, Gutierrez C, Laneelle MA, Swings J, Meyers WM, Daffe M, Portaels F. Characterization of an unusual mycobacterium: a possible missing link between Mycobacterium marinum and Mycobacterium ulcerans. J Clin Microbiol 2002; 40: 2370-2380.
Debacker M, Zinsou C, Aguiar J, Meyers W, Portaels F. Mycobacterium ulcerans disease (Buruli ulcer) following human bite [clinical picture]. Lancet 2002; 360: 1830.
Donners H, Willems B, Beirnaert E, Colebunders R, Davis D, van der Groen G. Cross-neutralizing antibodies against primary isolates in African women infected with HIV-1 [letter]. AIDS 2002; 16: 501-503.
Espey DK, Djomand G, Diomande I, Dosso M, Saki MZ, Kanga JM, Spiegel RA, Marston BJ, Gorelkin L, Meyers WM, Portaels F, Deming MS, Horsburgh CR. A pilot study of treatment of Buruli ulcer with rifampicin and dapsone. Int J Infect Dis 2002; 6: 60-65.
Florence E, Dreezen C, Desmet P, Smets E, Fransen K, Vandercam B, Pelgrom J, Clumeck N, Colebunders R. Ritonavir/saquinavir plus one nucleoside reverse transcriptase inhibitor (NRTI) versus indinavir plus two NRTIs in protease inhibitor-naive HIV-1-infected adults (IRIS study). Antiviral Ther 2002; 6: 255-262.
Ghys PD, Diallo MO, Ettiègne-Traoré V, Kalé K, Tawil O, Caraël M, Traoré M, Mah-bi G, De Cock KM, Wiktor SZ, Laga M, Greenberg AE. Increase in condom use and decline in HIV and sexually transmitted diseases among female sex workers in Abidjan, Côte d’Ivoire, 1991-1998. AIDS 2002; 16: 251-258.
Gómez-Marin JE, León Franco CI, Inirida Guerrero M, Rigouts L, Portaels F. IS6110 fingerprinting of sensitive and resistant strains (1991-1992) of Mycobacterium tuberculosis in Colombia. Mem Inst Oswaldo Cruz 2002; 97: 1005-1008.
Hargreaves JR, Morison LA, Chege J, Rutenberg N, Kahindo M, Weiss HA, Hayes R, Buvé A. Socioeconomic status and risk of HIV infection in an urban population in Kenya. Trop Med Int Health 2002; 7: 793-802.
Jennes W, Kestens L, Nixon DF, Shacklett BL. Enhanced ELISPOT detection of anti-genspecific T cell responses from cryopreserved specimens with addition of both IL-7 and IL-15: the Amplispot assay. J Immunol Methods 2002; 270: 99-108.
Jennes W, Sawadogo S, Koblavi-Dème S, Vuylsteke B, Maurice C, Roels TH, Chorba T, Nkengasong JN, Kestens L. Positive association between beta-chemokine-producing T cells and HIV type 1 viral load in HIV-infected subjects in Abidjan, Côte d’Ivoire. AIDS Res Hum Retroviruses 2002; 18: 171-177.
Johnson RC, Ifebe D, Hans-Moevi A, Kestens L, Houessou R, Guédénon A, Meyers WM, Portaels F. Disseminated Mycobacterium ulcerans disease in an HIV-positive patient: a case study [letter]. AIDS 2002; 16: 1704-1705.
Lagarde E, Caraël M, Auvert B, Buvé A. Concurrency and sexual transmission: a response to the letter by Rothenberg et al. [letter]. AIDS 2002; 16: 679-680.
Lowndes CM, Alary M, Meda H, Gnintoungbé CAB, Mukenge-Tshibaka L, Adjovi C, Buvé A, Morison L, Laourou M, Kanhonou L, Anagonou S. Role of core and bridging groups in the transmission dynamics of HIV and STIs in Cotonou, Benin, West Africa. Sex Transm Infect 2002; 78(Suppl.1): i69-i77.
Mijs W, de Haas P, Rossau R, Van der Laan T, Rigouts L, Portaels F, van Soolingen D. Molecular evidence to support a proposal to reserve the designation Mycobacterium avium subsp. avium for bird-type isolates and‘M. avium subsp. hominissuis’ for the human/porcine type of M. avium. IntJ Syst Evol Microbiol 2002; 52: 1505-1518.
Mijs W, De Vreese K, Devos A, Pottel H, Valgaeren A, Evans C, Norton J, Parker D, Rigouts L, Portaels F, Reischl U, Watterson S, Pfyffer G, Rossau R. Evaluation of a commercial line probe assay for identification of Mycobacterium species from liquid and solid culture. Eur J Clin Microbiol Infect Dis 2002; 21: 794-802.
Mukenge-Tshibaka L, Alary M, Lowndes CM, Van Dyck E, Guédou A, Geraldo N, Ana-gonou S, Lafia E, Joly JR. Syndromic versus laboratory-based diagnosis of cervical infections among female sex workers in Benin. Sex Transm Dis 2002; 29: 324-330.
Nyambi P, Heyndrickx L, Vereecken K, Burda S, De Houwer K, Coppens S, Urbanski M, Williams C, Ndumbe P, Janssens W. Predominance of infection with HIV-1 circulating recombinant form CRF02_AG in major Cameroonian cities and towns [letter]. AIDS 2002; 16: 295-296.
Ondoa P, Davis D, Kestens L, Vereecken C, Garcìa Ribas S, Fransen K, Heeney J, van der Groen G. In vitro susceptibility to infection with SIVcpz and HIV-1 is lower in chimpanzee than in human peripheral blood mononuclear cells. J Med Virol 2002; 67: 301-311.
Ondoa P, Vingerhoets J, Vereecken C, van der Groen G, Heeney JL, Kestens L. In vitro repli-cation of SIVcpz is suppressed by beta-chemokines and CD8+ T cells but not by natural killer cells of infected chimpanzees. AIDS Res Hum Retroviruses 2002; 18: 373-382.
Palomino JC, Martin A, Camacho M, Guerra H, Swings J, Portaels F. Resazurin microtiter assay plate: simple and inexpensive method for detection of drug resistance in Mycobacterium tuberculosis. Antimicrob Agents Chemother 2002; 46: 2720-2722.
Pattyn S, Grillone S. Relapse rates and a 10-year follow-up of a 6-week quadruple drug regimen for multibacillary leprosy. Lepr Rev 2002; 73: 245-247.
Portaels F, Aguiar J, Debacker M, Steunou C, Zinsou C, Guédénon A, Meyers WM. Prophylactic effect of Mycobacterium bovis BCG vaccination against osteomyelitis in children with Mycobacterium ulcerans disease (Buruli ulcer). Clin Diagn Lab Immunol 2002; 9: 1389-1391.
Scientific considerations for the regulation and clinical evaluation of HIV/AIDS preventive vaccines; report from a WHO-UNAIDS Consultation 13-15 March 2001, Geneva, Switzerland. AIDS 2002; 16: W15-W25.
Servais J, Hainaut M, Schmitz V, Maes P, Fransen K, Vaira D, Brichard B, Arendt V, Schneider F, Hemmer R, Schmit JC. Resistance testing in children changing human immunodeficiency virus type 1 protease inhibitor. Pediatr Infect Dis J 2002; 21: 214-220.
Van Damme L, Jespers V, Van Dyck E, Chapman A. Penile application of dextrin sulphate gel (Emmelle). Contraception 2002; 66: 133-136.
Van Damme L, Ramjee G, Alary M, Vuylsteke B, Chandeying V, Rees H, Sirivongrangson P, Mukenge-Tshibaka L, Ettiègne-Traoré V, Uaheowitchai C, Abdool Karim SS, Mâsse B, Perriëns J, Laga M. Effectiveness of COL-1492, a nonoxynol-9 vaginal gel, on HIV-1 transmission in female sex workers: a randomised controlled trial. Lancet 2002; 360: 971-977.
Van Deun A. Periodic or routine susceptibility testing in tuberculosis control [reply]. Int J Tuberc Lung Dis 2002; 6: 177-179.
Van Deun A, Salim AH, Cooreman E, Hossain A, Rema A, Chambugonj N, Hye A, Kawria A, Declercq E. Optimal tuberculosis case detection by direct sputum smear microscopy: how much better is more? Int J Tuberc Lung Dis 2002; 6: 222-230.
Van Herrewege Y, Penne L, Vereecken C, Fransen K, van der Groen G, Kestens L, Balzarini J, Vanham G. Activity of reverse transcriptase inhibitors in monocyte-derived dendritic cells: a possible in vitro model for postexposure prophylaxis of sexual HIV transmission. AIDS Res Hum Retroviruses 2002; 18: 1091-1102.
Watson-Jones D, Changalucha J, Gumodoka B, Weiss H, Rusizoka M, Ndeki L, Whitehouse A, Balira R, Todd J, Ngeleja D, Ross D, Buvé A, Hayes R, Mabey D. Syphilis in pregnancy in Tanzania. I. Impact of maternal syphilis on outcome of pregnancy. J Infect Dis 2002; 186: 940-947.
Watson-Jones D, Gumodoka B, Weiss H, Changalucha J, Todd J, Mugeye K, Buvé A, Kanga Z, Ndeki L, Rusizoka M, Ross D, Marealle J, Balira R, Mabey D, Hayes R. Syphilis in pregnancy in Tanzania. II. The effectiveness of antenatal syphilis screening and single-dose benzathine penicillin treatment for the prevention of adverse pregnancy outcomes. J Infect Dis 2002; 186: 948-957.
Zhong P, Burda S, Urbanski M, Kenfack H, Tongo M, Heyndrickx L, Nanfack A, Shang J, Agyingi L, Zolla-Pazner S, Zekeng L, Nyambi P. HIV type 1 group M clades infecting subjects from rural villages in Equatorial rain forests of Cameroon. J Acquir Immun Defic Syndr 2002; 31: 495-505.


Other publications and abstracts

Ackah AN, Akaki AA, Abouya YL, Traore M, Kamate M, Maurice C, Ekpini ER, Wiktor SZ, Laga M. Implementation of a program of cotrimoxazole prophylaxis to prevent opportunistic infections among tuberculosis patients with HIV infection attending outpatient TB clinics of Côte d’Ivoire [abstract]. In: XIV International AIDS Conference: “Knowledge and commitment for action”, Barcelona, July 7-12 2002; abstract book. [s.l.]: [s.n.], 2002: 262, Abstract Nr. WePeF6712.
Anoma C, Tiemélé A, Ettiègne-Traoré V, Koffi S, Vuylsteke B, Laga M. Limiting loss to follow-up rates during a microbicide trial: experiences during the COL-1492 trial in Abidjan, Côte d’Ivoire [abstract]. In: Microbicides, 12-15 May 2002, Antwerp, Belgium; conference abstracts. [s.l.]: [s.n.], 2002: 38, Abstract Nr. B-246.
Auer C, Montejo L, Lagahid J, Ang C, Van Deun A, Tanner M, Weiss MG. Randomized controlled trial of daily vs thrice weekly anti-TB therapy in public health centres of Manila, Philippines [abstract]. Int J Tuberc Lung Dis 2002; 6(Suppl.1): S149, Abstract Nr. 211-OP.
Ayokoin M, Vuylsteke B, Ettiègne-Traoré V, Coulibaly M, Danho-Bassimbié J, Tamany S. Advocacy to reinforce co-ordination of interventions targeted mobile and sexually vulnerable populations in Côte d’Ivoire [abstract]. In: XIV International AIDS Conference: “Knowledge and commitment for action”, Barcelona, July 7-12 2002; abstract book. [s.l.]: [s.n.], 2002: 226, -Abstract Nr. WePeE6573.
Buvé A. When RTC’s may not be the best design [abstract]. In: Microbicides, 12-15 May 2002, Antwerp, Belgium; conference abstracts. [s.l.]: [s.n.], 2002: 40, Abstract Nr. B-295.
Debacker M, Aguiar J, Meyers WM, Portaels F. Clinical and laboratory diagnosis of M. ulcerans disease [abstract]. Acta Trop 2002; 83(Suppl.1): S74, Abstract Nr. WEPS015.
Debacker M, Aguiar J, Steunou C, Zinsou C, Meyers WM, Guédénon A, Dramaix M, Portaels F. Trends in Mycobacterium ulcerans disease (Buruli ulcer) patients as seen in a rural hospital of southern Benin, 1997 through 2000 [abstract]. Acta Trop 2002; 83(Suppl.1): S74, Abstract Nr. WEPS014.
Delvaux T, Laga M, Buvé A. Integration of STI/HIV in reproductive health services [abstract]. In: Integration and disease control; international colloquium, Antwerp, 26-27-28 November 2002; abstract book. Antwerp: Institute of Tropical Medicine, 2002: 8.
Derdelinckx I, Van Laethem K, Maes B, Schrooten Y, Dewit S, Florence E, Fransen K, Ribas SG, Marissens D, Moutschen M, Van Wijngaerden E, Vaira D, Zissis G, Van Ranst M, Vandamme AM. Drug resistance among therapy-naive HIV-infected patients studied by sequencing and VERSANT (TM) HIV-1 resistance assays (LiPA) has limited impact on treatment response [abstract]. Antiviral Ther 2002; 7(Suppl.1): S181, Abstract Nr. 167.
Diomande F, Bissagnene E, Eholie S, Maurice C, Nkengasong J, Konan R, Kadio A, Laga M, Barouan MC. Towards simplified laboratory monitoring of patients on antiretroviral treatment (arv): experience from Côte d’Ivoire’s drug access initiative (dai) [abstract]. In: XIV International AIDS Conference: “Knowledge and commitment for action”, Barcelona, July 7-12 2002; abstract book. [s.l.]: [s.n.], 2002: 258, Abstract Nr. WePeF6694.
Donners H, Willems B, Heyndrickx L, Davis D, van der Groen G. ‘Extended incubation phase’ neutralization assays: insights for AIDS vaccine development [abstract]. J Hum Virol 2002; 5: 94, Abstract Nr. 264.
Donners H, Willems B, Heyndrickx L, Davis D, van der Groen G. Neutralization of HIV-1 primary isolates has a subtype specific component in assays quantifying a reduction in infectivity [abstract]. In: 7TH European Conference on Experimental AIDS Research (ECEAR 2002), Genoa, Italy June 8-11, 2002; book of abstracts. [s.l.]: [s.n.], 2002: 105, Abstract Nr. AB42.
Godfroid J, Pirard M, Fonteyne PA, Bughin J, Walravens K, Portaels F, Gala JL. Mortalities due to mycobacterial infections in wild red deer (Cervus elaphus) in Belgium [abstract]. In: 7th International Colloquium on Paratuberculosis, Bilbao, Spain, June 11-14, 2002. [s.l.]: [s.n.], 2002: 110.
Ir Por P, Van Damme W, Kheang ST, Coppé JAG, Ettema L, Crabbé F. Regular visits by female sex workers for STI control at special clinics are achievable on a voluntary basis; MSF’s experience in Cambodia [abstract]. In: XIV International AIDS Conference: “Knowledge and commitment for action”, Barcelona, July 7-12 2002; abstract book. [s.l.]: [s.n.], 2002: 234, Abstract Nr. WePpF2117.
Janssens W, Njai H, Ngong C, Van der Auwera G, van der Groen G. Development and evaluation of a probe hybridization assay for monitoring the prevalence of HIV-1 CRF02_AG in west and west central Africa [abstract]. In: 7TH European Conference on Experimental AIDS Research (ECEAR 2002), Genoa, Italy June 8-11, 2002; book of abstracts. [s.l.]: [s.n.], 2002: 53, Abstract Nr. WS30.
Jennes W, Vuylsteke B, Maurice C, Roels TH, Chorba T, Nkengasong J, Kestens L. Low levels of HIV-1-specific T helper cells in the peripheral blood of HIV-exposed seronegative female sex workers in Abidjan, Côte d’Ivoire [abstract]. In: XIV International AIDS Conference: “Knowledge and commitment for action”, Barcelona, July 7-12 2002; abstract book. [s.l.]: [s.n.], 2002: 358, Abstract Nr. ThPeA7096.
Jennes W, Vuylsteke B, Maurice C, Roels TH, Chorba T, Nkengasong J, Kestens L. Screening for HIV-1-specific T helper cell responses in the peripheral blood of HIV-exposed seronegative female sex workers in Abidjan, Côte d’Ivoire [abstract]. In: 7TH European Conference on Experimental AIDS Research (ECEAR 2002), Genoa, Italy June 8-11, 2002; book of abstracts. [s.l.]: [s.n.], 2002: 59, Abstract Nr. WS40.
Jespers V, Van Damme L, Chapman A. Penile application of dextrin sulphate gel (Emmelle) to HIV+VE and HIV-VE men [abstract]. In: Microbicides, 12-15 May 2002, Antwerp, Belgium; conference abstracts. [s.l.]: [s.n.], 2002: 38, Abstract Nr. B-235.
Johnson RC, Ifebe D, Hans-Moevi A, Kestens L, Houessou R, Guédénon A, Meyers WM, Portaels F. Disseminated Mycobacterium ulcerans disease in an HIV-positive patient: a case study [abstract]. Acta Trop 2002; 83(Suppl.1): S73, Abstract Nr.WEPS013.
Linden A, Canivet P, Mousset B, de Rijk P, Rigouts L, Portaels F. Evidence of paratuberculosis in wild red deer (Cervus elaphus) in Belgium [abstract]. In: 7th International Colloquium on Paratuberculosis, Bilbao, Spain, June 11-14, 2002. [s.l.]: [s.n.], 2002: 112.
Low-Beer N, Jespers V, McCormack S, Gabe R, Chapman A, Nunn AJ, Van Damme L, Lacey CJN. A dose-ranging I/II study of dextrin sulphate gel as a novel vaginal microbicide: data from 50 HIV-negative women [abstract]. Int J STD AIDS 2001; 12(Suppl.2): 36.
Low-Beer N, Jespers V, McCormack S, Gabe R, Chapman A, Nunn AJ, Van Damme L, Lacey CJN. A safety study of dextrin sulphate gel as a novel vaginal microbicide: data from HIV negative and positive women [abstract]. In: Microbicides, 12-15 May 2002, Antwerp, Belgium; conference abstracts. [s.l.]: [s.n.], 2002: 119, Abstract Nr. B-095.
Mean CV, Crabbé F, Godwin P. Managing HIV/AIDS interventions at national, provincial and operational district level in Cambodia [abstract]. In: Integration and disease control; international colloquium, Antwerp, 26-27-28 November 2002; abstract book. Antwerp: Institute of Tropical Medicine, 2002: 42.
Meyers WM, Aguiar J, Guédénon A, Debacker M, Maleombho-Usher M, Abalos FM, Portaels F. Spectrum of clinicopathologic features of Mycobacterium ulcerans disease (Buruli ulcer) [abstract]. Acta Trop 2002; 83(Suppl.1): S74, Abstract Nr. WEPS016.
Meyers WM, Portaels F. Mycobacterium ulcerans diseases (Buruli ulcer disease) [abstract]. In: Fifth WHO Advisory Group Meeting on Buruli Ulcer, March, 11-14, 2002, Geneva, Switzerland. [s.l.]: [s.n.], 2002.
Ngabonziza M, Lafort Y, Karita E. The methodology for evaluating STI case management in clinics in Rwanda [abstract]. In: XIV International AIDS Conference: “Knowledge and commitment for action”, Barcelona, July 7-12 2002; abstract book. [s.l.]: [s.n.], 2002: 274, Abstract Nr. MoPeF4046.
Njai HF, Janssens W, Van der Auwera G, De Vos E. Development of a probe hybridization assay to subtype human immunodeficiency virus-1 (HIV-1) circulatory recombinant form CRF02_AG(IbNg) [abstract]. Infection 2002; 30(Suppl.1): 21, Abstract Nr. 102.
Noba AV, Angoran H, Billy D, Maurice C, Coulibaly-Djeneba T, Mangle H, Tanoe JM, Shaffer N, Nolan M, Ekpini ER, Wiktor SZ, Laga M. Prevention of mother-to-child transmission of HIV (pmtct) including community mobilization and enhanced care and support: experience from Abidjan, Côte d’Ivoire [abstract]. In: XIV International AIDS Conference: “Knowledge and commitment for action”, Barcelona, July 7-12 2002; abstract book. [s.l.]: [s.n.], 2002: 266, Abstract Nr. WePeF6730.
Paranjothy S, Weiss H, Buvé A, Morison L. Factors associated with HIV infection among sex workers in four cities in sub-Saharan Africa [abstract]. In: XIV International AIDS Conference: “Knowledge and commitment for action”, Barcelona, July 7-12 2002; abstract book. [s.l.]: [s.n.], 2002: 133, Abstract Nr. MoPeC3493.
Portaels F, Aguiar J, Debacker M, Steunou C, Zinsou C, Guédénon A, Meyers WM. Prophylactic effect of BCG vaccination in children against osteomyelitis in Mycobacterium ulcerans disease (Buruli ulcer) [abstract]. Acta Trop 2002; 83(Suppl.1): S73, Abstract Nr. WEPS012.
Portaels F, Aguiar J, Debacker M, Steunou C, Zinsou C, Guédénon A, Meyers WM. Protective effect of BCG vaccination in children against severe forms of Mycobacterium ulcerans disease (Buruli ulcer) [abstract]. In: Fifth WHO Advisory Group Meeting on Buruli
Ulcer, March, 11-14, 2002, Geneva, Switzerland. [s.l.]: [s.n.], 2002.
Remes P, Kaona F, Kanhonou L, Njue C, Abega S. Urban African youth’s first and subsequent sexual experiences [abstract]. In: XIV International AIDS Conference: “Knowledge and commitment for action”, Barcelona, July 7-12 2002; abstract book. [s.l.]: [s.n.], 2002: 214, Abstract Nr. WePeE6519.
Rivera AB, Rigouts L, Grimaldo ER, Tupasi TE, Portaels F. The genetic diversity of multidrug resistant Mycobacterium tuberculosis from Filipino TB patients by spoligotyping [abstract]. In: Meeting of the Working Group on DOTS-Plus for MDR-TB, Tallinn, Estonia, 10-12 April 2002. [s.l.]: [s.n.], 2002.
Sajduda A, Dela A, Dziadek J, Augustynowicz-Kopec E, Portaels F. Molecular typing of drug-resistant Mycobacterium tuberculosis strains isolated in Poland during 2000 [abstract]. Int J Tuberc Lung Dis 2002; 6(Suppl.1): S197-S198, Abstract Nr. 257-PD.
Scott JT, Diakhaté M, Vereecken K, Fall A, Diop M, Ly A, Declercq D, de Vlas SJ, Berkvens D, Kestens L, Gryseels B. Age dependent resistance to Schistosoma mansoni infection; from water contact studies to cytokine-endocrine interactions [abstract]. Acta Trop 2002; 83(Suppl.1): S60, Abstract Nr.TUPS029.
Scott JT, Johnson CR, Guédénon A, Gryseels B, Portaels F. Buruli ulcer strikes independently of age, sex or Schistosoma haematobium infection in Benin [abstract]. Acta Trop 2002; 83(Suppl.1): S75, Abstract Nr. WEPS018.
Scott JT, Johnson CR, Guédénon A, Gryseels B, Portaels F. Risk factors for Buruli ulcer: age, sex and Schistosoma haematobium infection [abstract]. In: Fifth WHO Advisory Group Meeting on Buruli Ulcer, March, 11-14, 2002, Geneva, Switzerland. [s.l.]: [s.n.], 2002.
Van Deun A, Salim AH, Das PK, Bastian I, Portaels F. Outcome of a standardised regimen for multidrug-resistant tuberculosis in a resource-poor country [abstract]. Acta Trop 2002; 83(Suppl.1): S145, Abstract Nr. P099.
Van Herrewege Y, Penne L, Vereecken C, Van Roey J, De Bethune MP, Balzarini J, Kestens L, Vanham G. Microbicidal activity of dapivirine (TMC-120) and UC-781 in an in vitro model for prevention of sexual HIV transmission [abstract]. In: Microbicides, 12-15 May 2002, Antwerp, Belgium; conference abstracts. [s.l.]: [s.n.], 2002: 20, Abstract Nr. A-274.
Vandebosch A, Goetghebeur E, Van Damme L. Acceptability of vaginal gel as a protective barrier against HIV in African and Asian sex workers [abstract]. In: Microbicides, 12-15 May 2002, Antwerp, Belgium; conference abstracts. [s.l.]: [s.n.], 2002: 60, Abstract Nr. C-233.
Voisin C, Portaels F. Emergence of multidrug resistant tuberculosis in Eastern Europe. In: De Thé GB, Challoner D, Auquier L, editors. Confronting infections, antibiotic resistance and bioterrorism around the world; the role of Academies of Medicine; proceedings of the IAMP Conference, Paris 20-22 March 2002. [Paris]: Elsevier, 2002: 85-88.