A colorimetric assay for trypanosome viability and metabolic function.
We have adapted a tetrazolium salt (MTT) colorimetric cytotoxicity assay to the assessment of viability and metabolic function in cultured African trypanosomes. Trypomastigotes of Trypanosoma congolense and T. brucei rhodesianse were harvested from the blood of parasitemic rats and cultured under axenic conditions that support trypanosome viability and growth. Analysis of serial dilutions of these bloodstream forms indicated that the assay could detect 10(4) parasites. To assess the effect of lymphoid cytokines on trypomastigote viability, 10(5) freshly harvested parasites were cultured with a wide range of dilutions of human recombinant IL-1, IL-3, IL-6, interferon-gamma (IFN gamma) or tumor necrosis factor-alpha (TNF alpha), or bovine recombinant IFN gamma or TNF alpha for 24, 48 or 96 h. These cytokines had no apparent growth enhancing or inhibitory effect on the trypomastigotes compared with growth in supplemented medium alone. This assay has several advantages over traditional counting methods, including increased sensitivity and rapid, repeatable quantitation. This adaptation of the MTT colorimetric assay should be useful in screening drugs and host-derived factors for growth-modulating effects on trypanosomes and other extracellular protozoan parasites.
Ellis
J A
JA
International Laboratory for Research on Animal Diseases, Nairobi, Kenya.
Fish
W R
WR
Sileghem
M
M
McOdimba
F
F
D000818:Animals; D002417:Cattle; D003124:Colorimetry; D004396:Coloring Agents; D016207:Cytokines / Q000494:pharmacology; D008297:Male; D010084:Oxidation-Reduction; D051381:Rats; D013778:Tetrazolium Salts; D013844:Thiazoles; D016833:Trypanosoma brucei rhodesiense / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism; D014348:Trypanosoma congolense / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism
D004396:Coloring Agents; D016207:Cytokines; D013778:Tetrazolium Salts; D013844:Thiazoles; C022616:thiazolyl blue