Survey Protein Vaccine Formulated with Montanide ISA 70 Effects following Immunization and after Challenge with Leishmania major

N. Kh. Tehrani1, M. Mahdavi2, A.A. Imani Fooladi3 and F. Tabatabaie4*

1Biology Department, Science & Research Branch, Islamic Azad University, Tehran, Iran 2Virology Department, Pasteur Institute of Iran, Tehran, Iran. 3Applied Microbiology, Research Center, Baqiyatallah University of Medical Science, Teheran, Iran. 4Parasitology and Mycology Department, School of medicine, Iran University of Medical Sciences(IUMS),Tehran, Iran.

ABSTRACT:

Leishmaniasis is caused by parasitic protozoa of the genus Leishmania which, in the infected host are obliging intracellular parasite . Montanide ISA 70 is an adjuvant composed of a natural metabolizable oil and a highly refined emulsifier from the manide monooleate family. The TSA (thiol-specific antioxidant) is a considerable antigen of Leishmania major. The purpose of this work was Protein-vaccine efficacy in the presence Montanide comparing to absence of it. The expression of recombinant protein was confirmed with SDS (sodium dodecyl sulfate) page and Western bloting . 48 BALB/c mice were divided into four groups [TSA/Freund ,TSA/Alum+BCG , TSA/ Montanide and PBS groups] and immunized with 20 mg of vaccine subcutaneously three times intervals on days 0, 14 and 28. The mice were challenged with parasite 21 days after final immunization . The lymphocyte proliferation was evaluated with Brdu method. Cytokines was evaluated with ELISA method. The vaccine formulated with the recombinant TSA protein with Montanide induced lymphocytes proliferation , cytokines as compared with the control group. Base on results, current candidate vaccine has potency for futher studies.

KEYWORDS:

BCG; TSA; Montanide; Leishmaniasis; Vaccine; IFN-γ

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