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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.
Roum Arch Microbiol Immunol 2010 Jan-Mar; 69(1):5-12RA

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.

Authors+Show Affiliations

Department of Microbiology, Ilam University of Medical Sciences, Ilam, Iran. pakzad_i2006@yahoo.comNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

21053778

Citation

Pakzad, Iraj, et al. "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, vol. 69, no. 1, 2010, pp. 5-12.
Pakzad I, Rezaee A, Rasaee MJ, et al. 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. Roum Arch Microbiol Immunol. 2010;69(1):5-12.
Pakzad, I., Rezaee, A., Rasaee, M. J., Hossieni, A. Z., Tabbaraee, B., & Kazemnejad, A. (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, 69(1), pp. 5-12.
Pakzad I, et al. 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. Roum Arch Microbiol Immunol. 2010;69(1):5-12. PubMed PMID: 21053778.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - 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. AU - Pakzad,Iraj, AU - Rezaee,Abbas, AU - Rasaee,Mohammad J, AU - Hossieni,Ahmad Zavaran, AU - Tabbaraee,Bahman, AU - Kazemnejad,Anoshirvan, PY - 2010/11/9/entrez PY - 2010/11/9/pubmed PY - 2010/12/16/medline SP - 5 EP - 12 JF - Roumanian archives of microbiology and immunology JO - Roum Arch Microbiol Immunol VL - 69 IS - 1 N2 - 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. SN - 1222-3891 UR - https://www.unboundmedicine.com/medline/citation/21053778/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_ DB - PRIME DP - Unbound Medicine ER -