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Development of Luciferase Expressing Leishmania donovani Axenic Amastigotes as Primary Model for In Vitro Screening of Antileishmanial Compounds

Current Microbiology volume 65, pages 696–700 (2012)Cite this article

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.

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Acknowledgments

This work was supported by an in-house grant (SIP 0026) of the Central Drug Research Institute. We gratefully acknowledge CSIR for financial support to Ravinder. The manuscript carries CDRI communication no 8289.

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  1. Division of Biochemistry, CSIR-Central Drug Research Institute, Chattar Manzil Palace, Lucknow, 226001, UP, India

    Ravinder,  Bhaskar, Sonali Gangwar & Neena Goyal

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  1. Ravinder
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  2. Bhaskar
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  3. Sonali Gangwar
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  4. Neena Goyal
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Correspondence to Neena Goyal.

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Ravinder, Bhaskar, Gangwar, S. et al. Development of Luciferase Expressing Leishmania donovani Axenic Amastigotes as Primary Model for In Vitro Screening of Antileishmanial Compounds. Curr Microbiol 65, 696–700 (2012). https://doi.org/10.1007/s00284-012-0209-1

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  • DOI: https://doi.org/10.1007/s00284-012-0209-1

Keywords

  • Visceral Leishmaniasis
  • Leishmaniasis
  • Luciferase Expression
  • Miltefosine
  • Antileishmanial Activity