Biological and immunological characteristics of Brucella abortus S99 major outer membrane proteins
نویسندگان
چکیده
Introduction and objective: Outer membrane proteins (OMPs) of Brucella are considered as immunogenic structures which can be used to design and develop a subunit vaccine for human brucellosis. Brucella abortus S99 OMPs promote the synthesis of high levels of specific anti-Brucella IgG molecules in rabbits when administrated with lipopolysaccharide (LPS). The objective of this study is evaluation of the efficacy of B. abortus major OMPs with LPS in the induction of immune response against brucellosis. Materials and methods: OMPs were derived from B. abortus by sequential extraction of sonicated cells with ultracentrifugation and predigestion with lysozyme. Proteins could be separated by anion-exchange chromatography and gel-filtration. Based on SDS-PAGE profiles, porins have been dominantly purified among three different classes of B. abortus OMPs. Sera of immunized rabbits against B. abortus porins were analyzed by enzymelinked immunosorbent assay (ELISA). LPS of B. abortus and complete Freud's adjuvant (CFA) were also applied to elicit higher levels of anti-Brucella antibodies. Results: ELISA confirmed the potency of porins and porins combination with CFA and LPS to promote humoral specific response. Among the above-mentioned compounds, a combination of porins + LPS or porins + CFA has been the most potent immunogenic compound to induce higher titer of antibody against B. abortus S99 in the animal model. Conclusion: The application of a complex of Brucella LPS and porins as an effective method to elicit protective and long-lasting immunity against Brucella infection and would be studied to design and develop a subunit vaccine for human brucellosis.
منابع مشابه
ISOLATION AND PURIFICATION OF MAJOR OUTER MEMBRANE PROTEINS FROM BRUCELLA ABORTUS S-99
Isolation and purification of major outer membrane proteins (OMP) from the cell wall envelope of Brucella abortus S-99 were achieved by sonication, solubilization and membrane fractionation in the presence of non-ionic detergent (Tx-100) and lysozyme treatments, followed by ultracentrifugation. The crude OMP was treated with trypsin to free the preparation from any other protein contaminan...
متن کاملCloning and expression of Brucella outer membrane protein 36kDa (OMP2b) in E. coli
Background & Objective: Brucellosis is an important zoonotic disease of economic significance. Brucella species are gram-negative, facultative intracellular bacteria, and are capable of replicating in the phagosomes of macrophages. They cause infection in several animal species and humans. Prevention of new diseases and diagnosis of cases infected with the organism are both essential for eradic...
متن کاملProduction and Characterization of Monoclonal Antibodies against Brucella abortus S (99) Surface Antigens
By immunizing mice with killed whole bacterial cells of Brucella abortus S (99), a panel of six hybridomas producing monoclonal antibodies (mAb) specific for the surface antigens of this bacterium were produced. ELISA was used to screen the hybridoma supernatants. Immunoblots of the cell extract indicated that three mAb were specific for S-LPS (Ba-1, Ba-2, Ba-3) and three others were reactive w...
متن کاملBrucella outer membrane lipoprotein shares antigenic determinants with Escherichia coli Braun lipoprotein and is exposed on the cell surface.
In an enzyme-linked immunosorbent assay (ELISA), purified Brucella abortus and Escherichia coli peptidoglycan-linked lipoproteins gave a strong cross-reaction with sera from rabbits hyperimmunized with the heterologous lipoprotein. When smooth E. coli cells were used as ELISA antigens, the immunological cross-reaction was not observed unless the cells were treated to remove lipopolysaccharide a...
متن کاملEvaluation of immunogenicity and protective efficacy of a liposome containing Brucella abortus S19 outer membrane protein in BALB/c mice
The use of liposome as an adjuvant and a vaccine carrier has been cited previously in the literature. It has also been shown to be effective in enhancing the immunogenicity of vaccine candidates. BALB/c mice immunized subcutaneously with outer membrane protein (OMP) of Brucella abortus S19 vaccine strain entrapped in a commercial cationic liposome (S19-OMP-liposome) for vaccine delivery, showed...
متن کامل