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Differential gene expression analysis in antimony-unresponsive Indian kala azar (visceral leishmaniasis) clinical isolates by DNA microarray

Published online by Cambridge University Press:  19 February 2007

N. SINGH ,
R. ALMEIDA ,
H. KOTHARI ,
P. KUMAR ,
G. MANDAL ,
M. CHATTERJEE ,
S. VENKATACHALAM ,
M. K. GOVIND ,
S. K. MANDAL  and
S. SUNDAR
N. SINGH*
Affiliation:
Central Drug Research Institute, Lucknow, India
R. ALMEIDA
Affiliation:
Cambridge Institute for Medical Research, Wellcome Trust/MRC Building, Addenbrooke's Hospital, Hills Road, Cambridge
H. KOTHARI
Affiliation:
Central Drug Research Institute, Lucknow, India
P. KUMAR
Affiliation:
Central Drug Research Institute, Lucknow, India
G. MANDAL
Affiliation:
Department of Pharmacology, Institute of Post Graduate Medical Education and Research, Kolkata, India
M. CHATTERJEE
Affiliation:
Department of Pharmacology, Institute of Post Graduate Medical Education and Research, Kolkata, India
S. VENKATACHALAM
Affiliation:
Connexios Life Sciences, Bangalore, India
M. K. GOVIND
Affiliation:
Connexios Life Sciences, Bangalore, India
S. K. MANDAL
Affiliation:
Central Drug Research Institute, Lucknow, India
S. SUNDAR
Affiliation:
Banaras Hindu University, Varanasi, India
*
*Corresponding author: Central Drug Research Institute, Lucknow, India. E-mail: [email protected]
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Summary

In this study, cDNA microarray analysis of a closely related species, Leishmania major, was used as a screening tool to compare antimonial-resistant and susceptible clinical isolates of Leishmania donovani in order to to identify candidate genes on the basis of antimony resistance. Clinically confirmed resistant isolate 39 and sensitive isolate 2001 were used in this study. Many differentially regulated genes were identified whose expression levels differ in sodium antimony gluconate (SAG)-treated patients. Interestingly, genes on the array, showing changes in expression of over 2-fold revealed the identity of ABC transporters, which are known determinants of drug resistance in laboratory mutants. The functionality of the transporters was validated by flow cytometry which, being biologically informative, provides direct clues to gene function. The results suggest that isolate 39 could have developed resistance by an increased multidrug resistance protein (MRP)-like pump. This study provides preliminary clues to the role of a thiol-dependent efflux system in antimonial resistant clinical isolates of Leishmania donovani.

Keywords

Leishmaniamicroarraydrug resistanceclinical isolatesflow cytometry
Type
Research Article
Information
Parasitology , Volume 134 , Issue 6 , June 2007 , pp. 777 - 787
Copyright
Copyright © Cambridge University Press 2007

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