Leishmania (Viannia) panamensis: an in vitro assay using the expression of GFP for screening of antileishmanial drug.
Promastigotes of Leishmania (Viannia) panamensis were successfully transfected with p6.5-egfp to express green fluorescent protein. The transfectants remained infective to macrophages, providing an in vitro model for screening antileishmanial drugs. This was demonstrated by flow cytometry of macrophage-associated GFP after exposure of infected cultures to known antileishmanial drugs, i.e. amphotericin B and glucantime. Fluorescence of GFP diminished progressively from infected cells with increasing drug concentrations used in both cases. The availability of this fluorescent assay for infection of macrophages by L. (V.) panamensis facilitates drug discovery program for the Viannia species, which differ significantly from those of the Leishmania subgenus.
Varela M
Rubén E
RE
Programa de Estudio y Control de Enfermedades Tropicales-PECET, Universidad de Antioquia, Calle 62 No 52-59, Lab. 632, Medellín, Colombia.
Muñoz
Diana Lorena
DL
Robledo
Sara M
SM
Kolli
Bala K
BK
Dutta
Sujoy
S
Chang
Kwang Poo
KP
Muskus
Carlos
C
D000666:Amphotericin B / Q000494:pharmacology; D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D005434:Flow Cytometry; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D018334:Leishmania guyanensis / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D049408:Luminescent Agents / Q000378:metabolism*; D008264:Macrophages / Q000469:parasitology; D008536:Meglumine / Q000494:pharmacology; D000077485:Meglumine Antimoniate; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D009942:Organometallic Compounds / Q000494:pharmacology; D014162:Transfection; D020298:U937 Cells
D000981:Antiprotozoal Agents; D049408:Luminescent Agents; D009942:Organometallic Compounds; D049452:Green Fluorescent Proteins; D008536:Meglumine; D000077485:Meglumine Antimoniate; D000666:Amphotericin B