title: Development of luciferase expressing Leishmania donovani axenic amastigotes as primary model for in vitro screening of antileishmanial compounds.
abstract: The development of new therapeutic leads against leishmaniasis relies primarily on screening of a large number of compounds on multiplication of clinically irrelevant transgenic promastigotes. The advent of the successful in vitro culture of axenic amastigotes allows the development of transgenic axenic amastigotes as a primary screen which can test compounds in a high throughput mode like promastigotes, still representative of the clinically relevant mammalian amastigotes stage. The present study reports the development of luciferase-tagged axenic amastigotes of Leishmania donovani, the causative agent of Indian Kala-azar, for in vitro drug screening. Luciferase expressing promastigotes were transformed to axenic amastigotes at a low pH and high temperature without the loss of luciferase expression. As compared to transgenic promastigotes, the luciferase expressing axenic amastigotes exhibited more sensitivity to antileishmanial drugs, particularly to pentavalent antimony (~2.8-fold) and also to the test compounds. Hence, the developed luciferase expressing axenic amastigotes make an ideal choice for high throughput drug screening for antileishmanial compounds.
authors.affiliation: Division of Biochemistry, CSIR-Central Drug Research Institute, Chattar Manzil Palace, Lucknow 226001, UP, India.
authors.lastname: Ravinder
authors.lastname: Bhaskar
authors.forename: Sonali
authors.initials: S
authors.lastname: Gangwar
authors.forename: Neena
authors.initials: N
authors.lastname: Goyal
mesh_terms: D000818:Animals; D000965:Antimony / Q000494:pharmacology; D000967:Antimony Sodium Gluconate / Q000494:pharmacology; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D003470:Culture Media; D004353:Drug Evaluation, Preclinical; D006801:Humans; D007893:Leishmania donovani / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics / Q000254:growth & development*; D007898:Leishmaniasis, Visceral / Q000469:parasitology; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D021261:Parasitic Sensitivity Tests
chemical_list: D000981:Antiprotozoal Agents; D003470:Culture Media; D000965:Antimony; D008156:Luciferases; D000967:Antimony Sodium Gluconate