Immunological responses to recombinant cysteine synthase A of Brucella abortus in BALB/c mice
- 262 Downloads
Brucellosis is a worldwide zoonotic disease. No Brucella vaccine is available for use in humans, and existing animal vaccines have limitations. There is a need to develop a safe and effective vaccine against human and animal brucellosis. In the present study, we generated recombinant cysteine synthase A (rCysK) of Brucella abortus in Escherichia coli and purified it up to homogeneity by metal affinity chromatography. The immunogenicity and protective efficacy of purified rCysK were evaluated in BALB/c mice with Freund’s adjuvant, aluminium hydroxide gel or without any adjuvant. High titres of anti-rCysK IgG antibody predominated by IgG1 were observed in all immunized mice. After stimulation with rCysK, the spleen lymphocytes of mice immunized with CysK formulated with aluminium hydroxide gel produced significant levels of IFN-γ. Protection against challenge with virulent B. abortus strain 544 was determined in BALB/c mice, and significant protection was observed in all CysK immunized groups when compared with PBS control. Among all the CysK vaccine groups, comparatively better protection was observed in mice immunized with aluminium hydroxide gel (1.064 units of protection). Overall, the results of the study suggest that rCysK induces primarily Th2 type of immune response and provides partial protection against B. abortus challenge.
KeywordsCysteine synthase A Brucella Subunit vaccine IFN-gamma
The authors are thankful to Director, DRDE, for providing all facilities and support required for this study.
- Araya LN, Winter AJ (1990) Comparative protection of mice against virulent and attenuated strains of Brucella abortus by passive transfer of immune T cells or serum. Infect Immun 58:254–256Google Scholar
- Roitt J, Brostoff J, Male D (2001) Immunology. Mosby Publications, LondonGoogle Scholar
- Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, Vol. 1, 3rd edn. CSHL press, New YorkGoogle Scholar
- Stevens MG, Hennager SG, Olsen SC, Cheville NF (1994) Serologic responses in diagnostic tests for brucellosis in cattle vaccinated with Brucella abortus 19 or RB51. J Clin Microbiol 32:1065–1066Google Scholar
- Wu R, Sun Z, Wu J, Meng H, Zhang H (2010) Effect of bile salts stress on protein synthesis of Lactobacillus casei Zhang revealed by 2-dimensional gel electrophoresis. J Dairy Sci 93:3858–3868Google Scholar