Leishmania (Viannia) panamensis: An in vitro assay using the expression of GFP for screening of antileishmanial drug |
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| Authors: | Rubé n E. Varela M,Diana Lorena Muñ oz,Sara M. Robledo,Bala K. Kolli,Kwang Poo Chang |
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| Affiliation: | a Programa de Estudio y Control de Enfermedades Tropicales-PECET, Universidad de Antioquia, Calle 62 No 52-59, Lab. 632, Medellín, Colombia
b Department of Microbiology and Immunology, Chicago Medical School, Rosalind Franklin University, 3333 Green Bay Road, North Chicago, IL 60064, USA |
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| Abstract: | 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. |
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| Keywords: | L. (V.) panamensis L. (L.) amazonensis L. (L.) major L. (L.) donovani Viannia subgenus Leishmania subgenus Leishmaniasis Antileishmanial drug screening GFP, green fluorescent protein HIFBS, heat-inactivated fetal bovine serum PMA, phorbol myristate acetate PE, phycoerythrin nagt, N-acetylglucosamine-1-phosphate transferase gene |
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