A COLORIMETRIC ASSAY FOR TRYPANOSOME VIABILITY AND METABOLIC FUNCTION

被引:27
作者
ELLIS, JA
FISH, WR
SILEGHEM, M
MCODIMBA, F
机构
[1] UNIV SASKATCHEWAN,WESTERN COLL VET MED,DEPT VET MICROBIOL,SASKATOON S7N 0W0,SASKATCHEWAN,CANADA
[2] INT LAB RES ANIM DIS,NAIROBI,KENYA
关键词
D O I
10.1016/0304-4017(93)90015-F
中图分类号
R38 [医学寄生虫学]; Q [生物科学];
学科分类号
07 ; 0710 ; 09 ; 100103 ;
摘要
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 (IFNgamma) or tumor necrosis factor-alpha (TNFalpha), or bovine recombinant IFNgamma or TNFalpha 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.
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页码:143 / 149
页数:7
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