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Oliver Balmer

Publications and source records attributed to Oliver Balmer.

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New developments in human African trypanosomiasis.

PURPOSE OF REVIEW: To review recent literature on human African trypanosomiasis, focussing on genome sequencing, diagnosis and drug discovery, and typing of trypanosomes. RECENT FINDINGS: The most important recent development has been the completion of the Trypanosoma brucei genome which will greatly facilitate the discovery of new drug targets and genetic markers. Correct staging of the disease is of key importance for treatment. The analysis of sleep patterns is a promising new method to this end and has advanced enough to begin thorough clinical trials. In terms of novel drug candidates, dicationic molecules show the most promise with one oral diamidine in phase 3 clinical trials. New targets and classes of molecules which show in vitro trypanocidal activity are also described. Two new methods - MGE-PCR and microsatellites - allow analyses without parasite cultivation, eliminating a major impediment to efficient sampling for population studies. The finding that several wild animal species harbour T. b. gambiense, and that parasite transmission is efficient even from very low parasitaemias, sheds a new light on the importance of animal reservoirs. SUMMARY: The use of T. brucei as model system for molecular and cell biology is regularly producing new technologies exploitable for diagnosis and new drugs. Drug discovery and development experience a revival through new public-private partnerships and initiatives. The challenge remains to translate this progress into improvements for affected people in disease endemic areas.

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New fluorescence markers to distinguish co-infecting Trypanosoma brucei strains in experimental multiple infections.

Multiple-genotype infections are increasingly recognized as important factors in disease evolution, parasite transmission dynamics, and the evolution of drug resistance. However, the distinction of co-infecting parasite genotypes and the tracking of their dynamics have been difficult with traditional methods based on various genotyping techniques, leaving most questions unaddressed. Here we report new fluorescence markers of various colours that are inserted into the genome of Trypanosoma brucei to phenotypically label live parasites of all life cycle stages. If different parasite strains are labelled with different colours they can be easily distinguished from each other in experimental studies. A total of 10 T. brucei strains were successfully transfected with different fluorescence markers and were monitored in culture, tsetse flies and mice, to demonstrate stability of marker expression. The use of fluorescence activated cell sorting (FACS) allowed rapid and accurate identification of parasite strains labelled with different markers. Cell counts by FACS were virtually identical to counts by traditional microscopy (n=75, Spearman's rho: 0.91, p<0.0001) but were considerably faster and had a significantly lower sampling error (66% lower, d.f.=73, t=-17.1, p<0.0001). Co-infecting strains transfected with fluorescence genes of different colour were easily distinguished by eye and their relative and absolute densities were reliably counted by FACS in experimental multiple infections in mice. Since the FACS can simultaneously determine the population sizes of differently labelled T. brucei strains or subspecies it allows detailed and efficient tracking of multiple-genotype infections within a single host or vector individual, enabling more powerful studies on parasite dynamics. In addition, it also provides a simple way to separate genotypes after experimental mixed infections, to measure responses of the single strains to an applied treatment, thus eliminating the need for laborious cloning steps. The markers presented broaden the spectrum of tools available for experimental studies on multiple-genotype infections. They are fundamentally different from isoenzyme analysis and other genotyping approaches in that they allow the distinction of parasite genotypes based on an easily recognizable phenotypic trait. They will be of specific interest to researches addressing ecological, evolutionary and epidemiological questions using trypanosomes as an experimental system.

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