Cellular Markers for the Identification of Chemoresistant Isolates in Leishmania.

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