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Parasitology Research

, Volume 111, Issue 1, pp 223–230 | Cite as

Biomarkers of antimony resistance: need for expression analysis of multiple genes to distinguish resistance phenotype in clinical isolates of Leishmania donovani

  • Dhiraj Kumar
  • Ruchi Singh
  • Vasundhra Bhandari
  • Arpita Kulshrestha
  • Narendra Singh Negi
  • Poonam SalotraEmail author
Original Paper

Abstract

Resistance to antimony is a major cause of failure to therapy in a large proportion of visceral leishmaniasis cases. Methods to distinguish resistant and sensitive parasite are urgently needed as the standard in vitro intracellular drug susceptibility assays are cumbersome and time consuming. Differential expression profiling studies have led to the identification of several antimony resistance-associated genes; however, their efficacy as a potential biomarker for monitoring antimony resistance remains imprecise. We analysed the expression of eight genes [antimony metabolism-associated genes—multidrug resistance protein A (MRPA), γ-glutamylcysteine synthetase (γ-GCS) and aquaporin-1 (AQP1)—and genes identified by proteome/transcriptome profiling—heat shock protein 83, mitogen-activated protein kinase 1 and histones H1, H2A and H4) in antimony-resistant (n = 10) and antimony-sensitive (n = 4) clinical isolates of Leishmania donovani by quantitative real-time PCR, in comparison with a lab-generated resistant and a standard sensitive isolate. We observed a significant differential expression of MRPA, histone H1 (p < 0.01), γ-GCS, HSP83 (p < 0.005) and histone H2A and H4 (p < 0.0001) in a group of sodium antimony gluconate-resistant isolates compared to sensitive isolates. Preferential AQP1 expression was observed in all the sensitive isolates (p < 0.0001). Overall, expression profile in field isolates for all the genes studied showed altered expression in majority of isolates, while in some, the expression was static. All the isolates showed a mosaic of expression pattern of the genes analysed indicating constellation of genes contributes towards the drug susceptibility of parasite. As none of the genes exhibit an absolute correlation with phenotype, targeted expression analysis of a set of genes should be considered as biomarker for distinguishing the antimony-resistant and antimony-sensitive parasite.

Keywords

Antimony Visceral Leishmaniasis Field Isolate Resistant Isolate Drug Susceptibility 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Abbreviations

AQP1

Aquaporin-1

γ-GCS

γ-Glutamylcysteine synthetase

HSP83

Heat shock protein 83

MAPK1

Mitogen-activated protein kinase 1

MRPA

Multidrug resistance protein A

SAG

Sodium antimony gluconate

Notes

Acknowledgements

The authors are thankful to Jose M. Requena (Universidad Autonoma de Madrid, Spain) for anti-H2A antibodies. This work formed a part of PhD thesis of DK, submitted to BITS-Pilani, Rajasthan. Financial support of Indian Council of Medical Research, India is gratefully acknowledged. RS is a recipient of UNESCO L`oreal for Women in Science fellowship grant. DK and AK are grateful to the Council of Scientific and Industrial Research, India for fellowship.

Conflicts of interest

None to declare.

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Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Dhiraj Kumar
    • 1
    • 4
  • Ruchi Singh
    • 2
  • Vasundhra Bhandari
    • 1
  • Arpita Kulshrestha
    • 1
  • Narendra Singh Negi
    • 3
  • Poonam Salotra
    • 1
    Email author
  1. 1.National Institute of Pathology, Indian Council of Medical ResearchSafdarjung Hospital CampusNew DelhiIndia
  2. 2.National Institute of Malaria ResearchNew DelhiIndia
  3. 3.Department of MedicineSafdarjung HospitalNew DelhiIndia
  4. 4.Division of Emerging and Transfusion Transmitted DiseasesOffice of Blood Research and Review, CBER, FDAKensington BethesdaUSA

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