Resistance to pentamidine in Leishmania mexicana involves exclusion of the drug from the mitochondrion.
The uptake of [(3)H]pentamidine into wild-type and drug-resistant strains of Leishmania mexicana was compared. Uptake was carrier mediated. Pentamidine-resistant parasites showed cross-resistance to other toxic diamidine derivatives. A substantial decrease in accumulation of the drug accompanied the resistance phenotype, although the apparent affinity for pentamidine by its carrier was not altered when initial uptake velocity was measured. The apparent V(max), however, was reduced. An efflux of pentamidine could be measured in both wild-type and resistant cells. Only a relatively small proportion of the total accumulated pentamidine was available for efflux in wild-type cells, while in resistant cells the majority of loaded pentamidine was available for release. Pharmacological reagents which diminish the mitochondrial membrane potential reduced pentamidine uptake in wild-type parasites, and the mitochondrial membrane potential was shown to be reduced in resistant cells. A fluorescent analogue of pentamidine, 4',6'-diamidino-2-phenylindole, accumulated in the kinetoplast of wild-type but not resistant parasites. These data together indicate that diamidine drugs accumulate in the Leishmania mitochondrion and that the development of the resistance phenotype is accompanied by lack of mitochondrial accumulation of the drug and its exclusion from the parasites.
Basselin
Mireille
M
Division of Infection & Immunity, Institute of Biomedical and Life Sciences, University of Glasgow, United Kingdom.
Denise
Hubert
H
Coombs
Graham H
GH
Barrett
Michael P
MP
D000818:Animals; D000981:Antiprotozoal Agents / Q000493:pharmacokinetics*; D018432:Drug Resistance, Multiple / Q000502:physiology*; D005434:Flow Cytometry; D007894:Leishmania mexicana / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism*; D010419:Pentamidine / Q000493:pharmacokinetics*; D013045:Species Specificity
D000981:Antiprotozoal Agents; D010419:Pentamidine