Leishmania (Viannia) panamensis: an in vitro assay using the expression of GFP for screening of antileishmanial drug

Exp Parasitol. 2009 Jun;122(2):134-9. doi: 10.1016/j.exppara.2009.02.012. Epub 2009 Mar 20.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amphotericin B / pharmacology
  • Animals
  • Antiprotozoal Agents / pharmacology*
  • Flow Cytometry
  • Gene Expression
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism*
  • Humans
  • Leishmania guyanensis / drug effects*
  • Leishmania guyanensis / genetics
  • Leishmania guyanensis / metabolism
  • Luminescent Agents / metabolism*
  • Macrophages / parasitology
  • Meglumine / pharmacology
  • Meglumine Antimoniate
  • Microscopy, Fluorescence
  • Microscopy, Phase-Contrast
  • Organometallic Compounds / pharmacology
  • Transfection
  • U937 Cells

Substances

  • Antiprotozoal Agents
  • Luminescent Agents
  • Organometallic Compounds
  • Green Fluorescent Proteins
  • Meglumine
  • Meglumine Antimoniate
  • Amphotericin B