Rapid fluorescent assay for screening drugs on Leishmania amastigotes.
A rapid fluorescent viability assay employing alamarBlue was optimized for use with Leishmania axenic amastigotes, the stage of the parasite responsible for disease pathology. The activity of two protein kinase inhibitors, Staurosporine and H-89, as well as Amphotericin B, on promastigotes and amastigotes of Leishmania donovani and Leishmania tropica was compared. Both protein kinase inhibitors inhibited promastigote growth at lower concentrations than amastigotes, while the GI(50) for Amphotericin B on both stages was similar. This assay only requires a limited number of axenic amastigotes (50,000 cells/well) and can be used to rapidly screen large chemical or natural product libraries for activity against amastigotes.
The Kuvin Centre for the Study of Infectious and Tropical Diseases, Hebrew University - Hadassah Medical School, P.O. Box 12272, Jerusalem 91220, Israel.
Orly
O
Shimony
Charles L
CL
Jaffe
D000666:Amphotericin B / Q000494:pharmacology; D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005453:Fluorescence; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry; D007546:Isoquinolines / Q000494:pharmacology; D007893:Leishmania donovani / Q000187:drug effects* / Q000254:growth & development*; D007895:Leishmania tropica / Q000187:drug effects* / Q000254:growth & development*; D016773:Leishmaniasis, Cutaneous / Q000469:parasitology; D007898:Leishmaniasis, Visceral / Q000469:parasitology; D010078:Oxazines / Q000737:chemistry; D021261:Parasitic Sensitivity Tests / Q000379:methods*; D019311:Staurosporine / Q000494:pharmacology; D013449:Sulfonamides / Q000494:pharmacology; D013997:Time Factors; D014966:Xanthenes / Q000737:chemistry
D000981:Antiprotozoal Agents; D004791:Enzyme Inhibitors; D007202:Indicators and Reagents; D007546:Isoquinolines; D010078:Oxazines; D013449:Sulfonamides; D014966:Xanthenes; C005843:resazurin; D000666:Amphotericin B; D019311:Staurosporine; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide