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Clinical and Vaccine Immunology, May 2008, p. 836-842, Vol. 15, No. 5
1071-412X/08/$08.00+0     doi:10.1128/CVI.00433-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Kinesin Motor Domain of Leishmania donovani as a Future Vaccine Candidate{triangledown}

Ayan Dey,1 Pawan Sharma,2 Naresh Singh Redhu,1,{dagger} and Sarman Singh1*

Department of Laboratory Medicine, All India Institute of Medical Sciences, New Delhi 110029, India,1 Immunology Group, International Centre for Genetic Engineering and Biotechnology, New Delhi 110067, India2

Received 2 November 2007/ Returned for modification 5 February 2008/ Accepted 9 March 2008

Visceral leishmaniasis (VL) is one of the important parasitic diseases, with approximately 350 million people at risk. Due to the nonavailability of an ideal drug, development of a safe, effective, and affordable vaccine could be a solution for control and prevention of this disease. The present study was carried out to examine the immunological potential of kinesin protein from the microtubule locus of Leishmania donovani as a suitable vaccine candidate. In silico analysis of this region revealed clusters of major histocompatibility complex class I and II binding epitopes in its motor domain region. A recombinant protein was expressed from this region and named rLvacc. The antigenicity and immunogenicity studies of this protein by Western blot analysis revealed that rLvacc is strongly recognized by sera from acute VL patients. To evaluate its immunogenicity, peripheral blood mononuclear cells from cured VL patients were separated, and a lymphocyte proliferation assay was carried out in the presence of rLvacc. After lymphocyte proliferation, the pooled culture supernatant was assayed for anti-rLvacc antibody titers using an enzyme-linked immunosorbent assay. The results showed that immunoglobulin G2 (IgG2) subtype antibodies were predominant, while IgG1 subtype antibodies were produced in very low titers. On the basis of these ex vivo preliminary findings, its immunogenicity was studied in BALB/c mice. Vaccination with the DNA construct generated a good cellular immune response with significant increases in gamma interferon and interleukin-2 (IL-2) cytokine levels (Th1), but no increase in IL-4 levels (Th2). Taken together, our findings suggest the kinesin motor domain region of L. donovani as a potential vaccine candidate against visceral leishmaniasis.

* Corresponding author. Mailing address: FAMS, Division of Clinical Microbiology, All India Institute of Medical Sciences, P.O. Box 4398, Ansari Nagar, New Delhi 110029, India. Phone: 91-11-2658-8484. Fax: 91-11-2658-8663. E-mail: sarman_singh{at}yahoo.com

{triangledown} Published ahead of print on 19 March 2008.

{dagger} Present address: Department of Immunology, Faculty of Medicine, University of Manitoba, Winnipeg, MB, Canada R3E0W3.

Clinical and Vaccine Immunology, May 2008, p. 836-842, Vol. 15, No. 5
1071-412X/08/$08.00+0     doi:10.1128/CVI.00433-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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