Repository of Research and Investigative Information

Repository of Research and Investigative Information

Ilam University of Medical Sciences

Protection of BALB/C mice against Brucella abortus 544 challenge by vaccination with combination of recombinant human serum albumin-l7/l12 (Brucella abortus ribosomal protein) and lipopolysaccharide

(2010) Protection of BALB/C mice against Brucella abortus 544 challenge by vaccination with combination of recombinant human serum albumin-l7/l12 (Brucella abortus ribosomal protein) and lipopolysaccharide. Roumanian archives of microbiology and immunology. pp. 5-12. ISSN 1222-3891 (Print) 1222-3891 (Linking)

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Official URL: https://www.ncbi.nlm.nih.gov/pubmed/21053778

Abstract

BACKGROUND: The immunogenic Brucella abortus ribosomal protein L7/L12 and Lipopolysaccharide (LPS) are promising candidate antigens for the development of subunit vaccines against brucellosis. OBJECTIVE: This study was aimed to evaluate the protection of combination of recombinant HSA-L7/L12 fusion protein with LPS in Balb/c mouse. MATERIALS AND METHODS: The recombinant HSA-L7/L12 fusion protein in Saccharomyces cerevisiae was expressed and purified by affinity chromatography column. LPS was extracted by n-butanol, purified by ultracentrifugation. BALB/c mouses were immunized in 9 groups with PBS, HSA, tHSA-L7/L12, L7/L12, LPS, LPS+ HSA, LPS+ tHSA-L7/L12, LPS+ L7/L12, B. abortus S19. ELISA, LTT tests and challenging two weeks after last injection were carried out. Bacterial count of spleen of immunized BALB/c mouse was done four weeks after challenging with virulent strain B. abortus 544. RESULTS: In ELISA test the specific antibodies of tHSA-L7/L12 exhibited a dominance of immunoglobulin IgG1 over IgG2a. LPS-HSA and tHSA-L7/L12 + LPS produced a significantly higher antibody titer than LPS alone and L7/L12+LPS (P < 0.05). The predominant IgG subtype for LPS and L7/L12+LPS were IgG3. However, tHSA-L7/L12+ LPS and LPS+ HAS elicited predominantly IgG1 and IgG3 subtypes. In addition, the tHSA-L7/L12 fusion protein and L7/L12 elicited a strong T-cell proliferative response upon restimulation in vitro with recombinant tHSA-L7/L12 and L7/L12, suggesting the induction of a cellular immunity response in vivo. However, there was no significant difference proliferative response in L7/L12 and tHSA-L7/L12 fusion protein (P > 0.05). The combination of tHSA-L7/L12 fusion protein with LPS and B. abortus S19 induce higher level of protection against challenge with the virulent strain B. abortus 544 in BALB/c mice than other groups (P = 0.005). CONCLUSIONS: The combination of tHSA-L7/L12 fusion protein with LPS had higher protective ability than LPS and fusion protein distinctly.

Item Type: Article
Keywords: Adjuvants, Immunologic/administration & dosage Animals Bacterial Proteins/administration & dosage/genetics/*immunology Brucella Vaccine/administration & dosage/*genetics/*immunology Brucella abortus/*immunology Brucellosis/*prevention & control Female Humans Injections, Intraperitoneal Lipopolysaccharides/administration & dosage/*immunology Mice Mice, Inbred BALB C Recombinant Fusion Proteins/administration & dosage/genetics/*immunology Ribosomal Proteins/administration & dosage/genetics/*immunology Serum Albumin/immunology Vaccination Vaccines, Synthetic/administration & dosage/genetics/immunology
Page Range: pp. 5-12
Journal or Publication Title: Roumanian archives of microbiology and immunology
Journal Index: Pubmed
Volume: 69
Number: 1
ISSN: 1222-3891 (Print) 1222-3891 (Linking)
Depositing User: مهندس مهدی شریفی
URI: http://eprints.medilam.ac.ir/id/eprint/1198

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