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Increased levels of thiols protect antimony unresponsive Leishmania donovani field isolates against reactive oxygen species generated by trivalent antimony

Published online by Cambridge University Press:  05 July 2007

G. MANDAL ,
S. WYLLIE ,
N. SINGH ,
S. SUNDAR ,
A. H. FAIRLAMB  and
M. CHATTERJEE
G. MANDAL
Affiliation:
Department of Pharmacology, Institute of Post Graduate Medical Education and Research, Kolkata, 244B Acharya JC Bose Road, Kolkata-700 020, India
S. WYLLIE
Affiliation:
Division of Molecular Microbiology and Biological Chemistry, Wellcome Trust Biocentre, University of Dundee, Dundee, Scotland, UK
N. SINGH
Affiliation:
Drug Target Discovery and Development Division, Central Drug Research Institute, Lucknow, India
S. SUNDAR
Affiliation:
Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India
A. H. FAIRLAMB
Affiliation:
Division of Molecular Microbiology and Biological Chemistry, Wellcome Trust Biocentre, University of Dundee, Dundee, Scotland, UK
M. CHATTERJEE*
Affiliation:
Department of Pharmacology, Institute of Post Graduate Medical Education and Research, Kolkata, 244B Acharya JC Bose Road, Kolkata-700 020, India
*
*Corresponding author: Department of Pharmacology, Institute of Postgraduate Medical Education and Research, 244B Acharya JC Bose Road, Kolkata-700 020, India. Tel: +91 33 2223 4135. Fax: +9133 2280 1807. E-mail: ilatim@vsnl.net
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Summary

The current trend of antimony (Sb) unresponsiveness in the Indian subcontinent is a major impediment to effective chemotherapy of visceral leishmaniasis (VL). Although contributory mechanisms studied in laboratory-raised Sb-R parasites include an up-regulation of drug efflux pumps and increased thiols, their role in clinical isolates is not yet substantiated. Accordingly, our objectives were to study the contributory role of thiols in the generation of Sb unresponsiveness in clinical isolates. Promastigotes were isolated from VL patients who were either Sb responsive (n=2) or unresponsive (n=3). Levels of thiols as measured by HPLC and flow cytometry showed higher basal levels of thiols and a faster rate of thiol regeneration in Sb unresponsive strains as compared with sensitive strains. The effects of antimony on generation of reactive oxygen species (ROS) in normal and thiol-depleted conditions as also their H2O2 scavenging activity indicated that in unresponsive parasites, Sb-mediated ROS generation was curtailed, which could be reversed by depletion of thiols and was accompanied by a higher H2O2 scavenging activity. Higher levels of thiols in Sb-unresponsive field isolates from patients with VL protect parasites from Sb-mediated oxidative stress, thereby contributing to the antimony resistance phenotype.

Keywords

visceral leishmaniasisantimonial resistancethiolsoxidative stressreactive oxygen species
Type
Research Article
Information
Parasitology , Volume 134 , Issue 12 , November 2007 , pp. 1679 - 1687
Copyright
Copyright © Cambridge University Press 2007

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