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A Parasite Rescue and Transformation Assay for Antileishmanial Screening Against Intracellular Leishmania donovani Amastigotes in THP1 Human Acute Monocytic Leukemia Cell Line
12:22

A Parasite Rescue and Transformation Assay for Antileishmanial Screening Against Intracellular Leishmania donovani Amastigotes in THP1 Human Acute Monocytic Leukemia Cell Line

Published on: December 30, 2012

Development and characterization of paromomycin-resistant Leishmania donovani promastigotes

M Maarouf1, M T Adeline, M Solignac

  • 1University of Damascus, Faculty of Pharmacy, Dept of Biochemistry & Microbiology, Syria.

Parasite (Paris, France)
|October 1, 1998
PubMed

Insights

Researchers developed paromomycin-resistant Leishmania donovani. This resistance was stable and did not affect parasite infectivity, suggesting altered drug uptake as the primary resistance mechanism.

Area of Science:

  • Parasitology
  • Molecular Biology
  • Drug Resistance

Background:

  • Paromomycin is a key chemotherapeutic agent against leishmaniasis.
  • Understanding drug resistance mechanisms is crucial for effective treatment.

Purpose of the Study:

  • To investigate the characteristics and mechanisms of paromomycin resistance in Leishmania donovani.

Main Methods:

  • Selection of resistant promastigotes through incremental drug pressure.
  • Cloning and characterization of resistant parasites.
  • Assessment of drug resistance stability in vitro and in vivo.
  • Development of HPLC method for intracellular drug quantification.

Main Results:

  • Stable paromomycin resistance (up to 800 microM) was achieved in Leishmania donovani promastigotes.
  • Resistant parasites did not exhibit multidrug resistance.
  • Resistance remained stable in the absence of paromomycin and in amastigotes after mouse passage.
  • Resistant parasites maintained infectivity for macrophages and mice.
  • Decreased intracellular paromomycin uptake, likely due to altered membrane composition, was identified as the primary resistance mechanism.

Conclusions:

  • Paromomycin resistance in Leishmania donovani is stable and does not compromise parasite infectivity.
  • Altered membrane composition leading to reduced drug uptake is the principal mechanism of resistance.
  • These findings have implications for managing leishmaniasis treatment strategies.

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