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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

Information

Type
Research Article
Information
Parasitology , Volume 134 , Issue 12 , November 2007 , pp. 1679 - 1687
Copyright
Copyright © Cambridge University Press 2007

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References

REFERENCES

Borst, P. and Ouellette, M. (1995). New mechanisms of drug resistance in parasitic protozoa. Annual Review of Microbiology 49, 427–460.CrossRefGoogle ScholarPubMed
Chakraborty, A. K. and Majumder, H. K. (1988). Mode of action of pentavalent antimonials: specific inhibition of type I DNA topoisomerase of Leishmania donovani. Biochemical and Biophysical Research Communications 152, 605–611.Google Scholar
Croft, S. L., Sundar, S. and Fairlamb, A. H. (2006). Drug resistance in leishmaniasis. Clinical Microbiology Reviews 19, 111–126.Google Scholar
Dey, S., Papadopoulou, B., Haimeur, A., Roy, G., Grondin, K., Dou, D., Rosen, B. P. and Ouellette, M. (1994). High level of arsenite resistance in Leishmania tarentolae is mediated by an active extrusion system. Molecular and Biochemical Parasitology 67, 49–57.CrossRefGoogle ScholarPubMed
Fairlamb, A. H. and Cerami, A. (1992). Metabolism and functions of trypanothione in the Kinetoplastida. Annual Review of Microbiology 46, 695–729.Google Scholar
Ghosh, S., Goswami, S. and Adhya, S. (2003). Role of superoxide dismutase in survival of Leishmania within the macrophage. The Biochemical Journal 369, 447–452.Google Scholar
Gottesman, M. M. (2002). Mechanisms of cancer drug resistance. Annual Review of Medicine 53, 615–627.CrossRefGoogle ScholarPubMed
Griffith, O. W. and Meister, A. (1979). Potent and specific inhibition of glutathione synthesis by buthionine sulfoximine (S-n-butyl homocysteine sulfoximine). Journal of Biological Chemistry 254, 7558–7560.Google Scholar
Grondin, K., Haimeur, A., Mukhopadhyay, R., Rosen, B. P. and Ouellette, M. (1997). Coamplification of the gamma-glutamylcysteine synthetase gene gsh1 and of the ABC transporter gene pgpA in arsenite resistant Leishmania tarentolae. EMBO Journal 16, 3057–3065.Google Scholar
Guerin, P. J., Olliaro, P., Sundar, S., Boelaert, M., Croft, S. L., Desjeux, P., Wasunna, M. K. and Bryceson, A. D. (2002). Visceral leishmaniasis: current status of control, diagnosis, and treatment, and a proposed research and development agenda. Lancet Infectious Diseases 2, 494–501.CrossRefGoogle Scholar
Haimeur, A., Brochu, C., Genest, P., Papadopoulou, B. and Ouellette, M. (2000). Amplification of the ABC transporter gene PGPA and increased trypanothione levels in potassium antimonyl tartrate (Sb III) resistant Leishmania tarentolae. Molecular and Biochemical Parasitology 108, 131–135.CrossRefGoogle Scholar
Haimeur, A., Guimond, C., Pilote, S., Mukhopadhyay, R., Rosen, B. P., Poulin, R. and Ouellette, M. (1999). Elevated levels of polyamines and trypanothione resulting from overexpression of the ornithine decarboxylase gene in arsenite resistant Leishmania. Molecular Microbiology 34, 726–735.CrossRefGoogle ScholarPubMed
Herwaldt, B. L. (1999). Leishmaniasis. Lancet 354, 1191–1199.Google Scholar
Jaffe, C. L., McMahon-Pratt, D. (1987). Serodiagnostic assay for visceral leishmaniasis employing monoclonal antibodies. Transactions of the Royal Society for Tropical Medicine and Hygiene 81, 587–594.CrossRefGoogle ScholarPubMed
Lee, C. W. (2004). Reversing agents for ATP-binding cassette (ABC) transporters: application in modulating multidrug resistance (MDR). Current Medicinal Chemistry Anti-Cancer Agents 4, 43–52.Google Scholar
Lu, S. C. (2000). Regulation of glutathione synthesis. Current Topics in Cellular Regulation 36, 95–116.Google Scholar
Marquis, N., Gourbal, B., Rosen, B. P., Mukhopadhyay, R. and Ouellette, M. (2005). Modulation in aquaglyceroporin AQP1 gene transcript levels in drug-resistant Leishmania. Molecular Microbiology 57, 1690–1699.CrossRefGoogle ScholarPubMed
Mehta, A. and Shaha, C. (2006). Mechanism of metalloid-induced death in Leishmania spp.: role of iron, reactive oxygen species, Ca2+, and glutathione. Free Radical Biology and Medicine 40, 1857–1868.CrossRefGoogle ScholarPubMed
Meister, A. and Anderson, M. E. (1983). Glutathione. Annual Review of Biochemistry 52, 711–760.Google Scholar
Mookerjee Basu, J., Mookerjee, A., Sen, P., Bhaumik, S., Sen, P., Banerjee, S., Naskar, K., Choudhuri, S. K., Saha, B., Raha, S. and Roy, S. (2006). Sodium antimony gluconate induces generation of reactive oxygen species and nitric oxide via phosphoinositide 3-kinase and mitogen-activated protein kinase activation in Leishmania donovani-infected macrophages. Antimicrobial Agents and Chemotherapy 50, 1788–1797.CrossRefGoogle ScholarPubMed
Mukherjee, A., Prasad, K. P., Singh, S., Roy, G., Girard, I., Chatterjee, M., Ouellette, M. and Madhubala, R. (2007). Role of ABC transporter MRPA, γ-glutamylcysteine synthetase and ornithine decarboxylase in natural antimony-resistant isolates of Leishmania donovani. Journal of Antimicrobial Chemotherapy 59, 204–211.Google Scholar
Mukhopadhyay, R., Dey, S., Xu, N., Gage, D., Lightbody, J., Ouellette, M. and Rosen, B. P. (1996). Trypanothione overproduction and resistance to antimonials and arsenicals in Leishmania. Proceedings of the National Academy of Sciences, USA 93, 10383–10387.Google Scholar
Murray, H. W., Berman, J. D., Davies, C. R. and Saravia, N. G. (2005). Advances in leishmaniasis. Lancet 366, 1576–1577.Google Scholar
O'Connor, J. E., Kimler, B. F., Morgan, M. C. and Tempas, K. J. (1988). A flow cytometric assay for intracellular nonprotein thiols using mercury orange. Cytometry 9, 529–532.Google Scholar
Ouellette, M. and Borst, P. (1991). Drug resistance and P-glycoprotein gene amplification in the protozoan parasite Leishmania. Research in Microbiology 142, 737–746.CrossRefGoogle ScholarPubMed
Roberts, W. L. and Rainey, P. M. (1993). Antileishmanial activity of sodium stibogluconate fractions. Antimicrobial Agents and Chemotherapy 37, 1842–1846.Google Scholar
Sereno, D., Cavaleyra, M., Zemzoumi, K., Maquaire, S., Ouaissi, A. and Lemesre, J. L. (1998). Axenically grown amastigotes of Leishmania infantum used as an in vitro model to investigate the pentavalent antimony mode of action. Antimicrobial Agents and Chemotherapy 42, 3097–3102.Google Scholar
Shaked-Mishan, P., Ulrich, N., Ephros, M. and Zilberstein, D. (2001). Novel intracellular Sbv reducing activity correlates with antimony susceptibility in Leishmania donovani. Journal of Biological Chemistry 276, 3971–3976.CrossRefGoogle ScholarPubMed
Shim, H. and Fairlamb, A. H. (1988). Levels of polyamines, glutathione and glutathione-spermidine conjugates during growth of the insect trypanosomatid Crithidia fasciculata. Journal of General Microbiology 134, 807–817.Google ScholarPubMed
Singh, N., Almeida, R., Kothari, H., Kumar, P., Mandal, G., Chatterjee, M., Venkatachalam, S., Govind, M. K., Mandal, S. K. and Sundar, S. (2007). Differential gene expression analysis in antimony unresponsive Indian kala azar (visceral leishmaniasis) clinical isolates by DNA microarray. Parasitology 134, 777–787.Google Scholar
Sundar, S. and Chatterjee, M. (2006). Visceral leishmaniasis – current therapeutic modalities. Indian Journal of Medical Research 123, 345–352.Google Scholar
TDR (2005). Leishmaniasis. Seventeenth Programme Report, Progress 2003–2004, 19–25.Google Scholar
Wan, C. P., Myung, E. and Lau, B. H. (1993). An automated micro-fluorometric assay for monitoring oxidative burst activity of phagocytes. Journal of Immunological Methods 159, 131–138.Google Scholar
Wyllie, S., Cunningham, M. L. and Fairlamb, A. H. (2004). Dual action of antimonial drugs on thiol redox metabolism in the human pathogen Leishmania donovani. Journal of Biological Chemistry 279, 39925–39932.CrossRefGoogle ScholarPubMed