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Cellular Markers for the Identification of Chemoresistant Isolates in Leishmania

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Trypanosomatids

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2116))

Abstract

Markers to diagnose chemoresistance in infecting Leishmania parasites are urgently required. This is fundamental for patients who do not heal during or after treatment, as they are unresponsive, or patients who relapse at the end of the therapy, suffering from therapeutic failure. Glucose utilization is an indicator of cell viability that closely associates with metabolic activity. In Leishmania, glucose is a source of carbon atoms and is imported into the cell through specific transporters. In experimentally developed chemoresistant Leishmania parasites a significant decrease of the expression of glucose transporters as well as in the cellular accumulation glucose has been described. Alternatively, the electrical membrane potential is an essential parameter for the formation of the electromotive force needed for the acquisition of important nutrients and solutes (e.g., glucose) by cells, and changes in glucose concentration are suggested to constitute a physiological adaptation associated with a chemoresistant phenotype of Leishmania parasites. Here we describe easy methods to measure glucose uptake and the membrane potential in isolates from patient suffering leishmaniasis. Correlation between both parameters might be helpful to identify chemoresistant parasites. Results suggest that the measured kinetics of glucose utilization rate can be correlated with the plasma membrane potential and together used to differentiate between the performance of wild-type and reference parasites on the one hand and parasites isolated from patients with therapeutic failure on the other.

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Acknowledgments

The authors are grateful to Dr. Emilia Diaz, Med. Vet. Wilmer Alcazar, and Med. Doc. Michelle Giammarressi for details related to the methods herein described and to Mrs. Pilar Rodríguez for her technical assistance. Likewise, they are grateful for the support conferred by the Alexander von Humboldt Foundation and the Siebold-Collegium Institute for Advanced Studies, University of Würzburg, Germany, to Alicia Ponte-Sucre. The authors are grateful to the Universidad Central de Venezuela Council for Research, grants CDCH-UCV PI-09-8717-2013/1 and PG-09-8646-2013/1. This project was approved by the Ethical Committee of the Institute of Biomedicine, Universidad Central de Venezuela.

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Authors and Affiliations

  1. Laboratorio de Fisiología Molecular, Facultad de Medicina, Instituto de Medicina Experimental, Universidad Central de Venezuela, Caracas, Venezuela

    Maritza Padrón-Nieves & Alicia Ponte-Sucre

Authors
  1. Maritza Padrón-Nieves
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  2. Alicia Ponte-Sucre
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Editors and Affiliations

  1. School of Biological Sciences, University of Edinburgh, Edinburgh, UK

    Paul A. M. Michels

  2. School of Applied Sciences, University of Huddersfield, Huddersfield, UK

    Michael L. Ginger

  3. Faculty of Biology, Technion–Israel Institute of Technology, Haifa, Israel

    Dan Zilberstein

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Padrón-Nieves, M., Ponte-Sucre, A. (2020). Cellular Markers for the Identification of Chemoresistant Isolates in Leishmania. In: Michels, P., Ginger, M., Zilberstein, D. (eds) Trypanosomatids. Methods in Molecular Biology, vol 2116. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0294-2_44

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  • DOI: https://doi.org/10.1007/978-1-0716-0294-2_44

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0293-5

  • Online ISBN: 978-1-0716-0294-2

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