Leishmania infantum promastigotes: flow cytometry as a possible tool for assessing the effects of drugs on cellular functions.
The capacity of flow-cytometric techniques to detect drug-specific biochemical targets and side effects in Leishmania infantum promastigotes was estimated by assessing the effects of three antileishmanial drugs (pentamidine, allopurinol, and amphotericin B) on parasite metabolism. Cell cycle and total protein content were estimated by staining cells with propidium iodide and fluorescein isothiocyanate, nonprotein thiols were stained by mercury orange, and membrane potential was measured by the accumulation of 3,3'-dipenthyloxacarbocyanine iodide inside the cell. Results showed that dynamic studies in parasites treated with subtoxic concentrations of drugs allowed the detection of drug-specific targets: pentamidine primarily affected nonprotein thiol contents and DNA synthesis, allopurinol primarily affected intracellular protein contents, and amphotericin B primarily affected membrane potential. Moreover, the assessment of cellular functions in parasites treated with increasing concentrations of drugs certified the capacity of these techniques to establish dose-response curves and to permit the detection of side effects.
Azas
N
N
Laboratoire de Parasitologie, Faculté de Pharmacie, Marseille, France.
Di Giorgio
C
C
Delmas
F
F
Gasquet
M
M
Timon-David
P
P
D000493:Allopurinol / Q000494:pharmacology*; D000666:Amphotericin B / Q000494:pharmacology*; D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D002232:Carbocyanines; D002453:Cell Cycle / Q000187:drug effects; D004396:Coloring Agents; D016054:DNA, Protozoan / Q000032:analysis / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D018314:Leishmania infantum / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D008564:Membrane Potentials / Q000187:drug effects; D010419:Pentamidine / Q000494:pharmacology*; D010663:Phenylmercury Compounds; D011419:Propidium; D015800:Protozoan Proteins / Q000032:analysis / Q000187:drug effects; D013438:Sulfhydryl Compounds / Q000032:analysis; D013439:Sulfhydryl Reagents
D000981:Antiprotozoal Agents; D002232:Carbocyanines; D004396:Coloring Agents; D016054:DNA, Protozoan; D005456:Fluorescent Dyes; D010663:Phenylmercury Compounds; D015800:Protozoan Proteins; D013438:Sulfhydryl Compounds; D013439:Sulfhydryl Reagents; D011419:Propidium; C018528:3,3'-dipentyl-2,2'-oxacarbocyanine; D000493:Allopurinol; D010419:Pentamidine; D000666:Amphotericin B; D016650:Fluorescein-5-isothiocyanate; C031504:mercury orange