Flow cytofluorimetric analysis of drug accumulation by multidrug-resistant Trypanosoma brucei brucei and T. b. rhodesiense.
Dual laser flow cytofluorimetry has been used to compare accumulation of compounds representing three major classes of trypanocidal drugs by drug sensitive and drug resistant clones of Trypanosoma brucei brucei and T. b. rhodesiense. Clones selected for resistance to melarsoprol were shown to be cross-resistant in vivo to two diamidines, pentamidine and Berenil, but not to suramin. At 35 degrees C, bloodstream forms of these multidrug-resistant clones accumulated lower intracellular concentrations of the diamidines 4',6-diamidino-2-phenyl-indole (DAPI) and Hoechst 33342, the phenanthridine ethidium bromide, and the acridine acriflavine than drug sensitive parasites. Accumulation of all four drugs was saturable. Drug concentrations giving half-maximal rates of accumulation were increased in the resistant clones relative to the sensitive parent clones. The rate of DAPI accumulation by both resistant and sensitive parasites was strongly temperature dependent and increased sharply above 27 degrees C. Two distinct populations were resolved in mixtures of sensitive and resistant clones exposed to DAPI. Resistant and sensitive cells accumulated identical intracellular concentrations of DAPI following brief treatment with the detergent Triton X-100. The results suggest that alterations in the surface membrane of multidrug-resistant trypanosomes reduce accumulation of several drugs relative to drug sensitive parasites.
Frommel
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Department of Microbiology and Immunology, Duke University Medical Center, Durham, NC 27710.
Balber
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D000167:Acriflavine / Q000378:metabolism; D000818:Animals; D001562:Benzimidazoles / Q000378:metabolism; D004133:Diminazene / Q000031:analogs & derivatives / Q000494:pharmacology; D004351:Drug Resistance; D004996:Ethidium / Q000378:metabolism; D005434:Flow Cytometry; D005456:Fluorescent Dyes / Q000378:metabolism*; D006801:Humans; D007211:Indoles / Q000378:metabolism; D008549:Melarsoprol / Q000494:pharmacology; D051379:Mice; D010419:Pentamidine / Q000494:pharmacology; D013498:Suramin / Q000494:pharmacology; D014344:Trypanocidal Agents / Q000378:metabolism* / Q000494:pharmacology; D014346:Trypanosoma brucei brucei / Q000187:drug effects* / Q000378:metabolism
D001562:Benzimidazoles; D005456:Fluorescent Dyes; D007211:Indoles; D014344:Trypanocidal Agents; D000167:Acriflavine; C007293:DAPI; D013498:Suramin; D010419:Pentamidine; D004996:Ethidium; C003915:diminazene aceturate; C017807:bisbenzimide ethoxide trihydrochloride; D004133:Diminazene; D008549:Melarsoprol