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World J Gastroenterol. Oct 28, 2008; 14(40): 6244-6248
Published online Oct 28, 2008. doi: 10.3748/wjg.14.6244
Immunogenicity and immunoprotection of recombinant PEB1 in Campylobacter-jejuni-infected mice
Lian-Feng Du, Zhen-Jiang Li, Xian-Ying Tang, Jun-Qiong Huang, Wan-Bang Sun
Lian-Feng Du, Zhen-Jiang Li, Xian-Ying Tang, Jun-Qiong Huang, Wan-Bang Sun, Department of Immunology, Zhuhai Campus of Zunyi Medical College, Zhuhai 519041, Guangdong Province, China
Author contributions: Sun WB and Du LF designed the research; Du LF, Tang XY and Li ZJ performed the research and data collection; Du LF and Huang JQ performed the analysis; Du LF and Sun WB wrote the paper.
Supported by Grants from the Governor Foundation for Excellent Talents of Guizhou Province, No. 200607
Correspondence to: Dr. Wan-Bang Sun, Professor, Department of Immunology, Zhuhai Campus of Zunyi Medical College, Zhuhai 519041, Guangdong Province, China. sunwb7224@sina.com
Telephone: +86-756-7623360 Fax: +86-756-7623388
Received: July 8, 2008
Revised: October 6, 2008
Accepted: October 13, 2008
Published online: October 28, 2008
Abstract

AIM: To construct a prokaryotic expression vector carrying Campylobacter jejuni peb1A gene and express it in Escherichia coli. Immunoreactivity and antigenicity of rPEB1 were evaluated. The ability of rPEB1 to induce antibody responses and protective efficacy was identified.

METHODS: peb1A gene was amplified by PCR, target gene and prokaryotic expression plasmid pET28a (+) was digested with BamHI and XhoI, respectively. DNA was ligated with T4 DNA ligase to construct recombinant plasmid pET28a(+)-peb1A. The rPEB1 was expressed in E. coli BL21 (DE3) and identified by SDS-PAGE. BALB/c mice were immunized with rPEB1. ELISA was used to detect the specific antibody titer and MTT method was used to measure the stimulation index of spleen lymphocyte transformation.

RESULTS: The recombinant plasmid pET28a (+)-peb1A was correctly constructed. The expression output of PEB1 protein in pET28a (+)-peb1A system was approximately 33% of total proteins in E. coli. The specific IgG antibody was detected in serum of BALB/c mice immunized with rPEB1 protein. Effective immunological protection with a lower sickness incidence and mortality was seen in the mice suffering from massive C. jejuni infection.

CONCLUSION: rPEB1 protein is a valuable candidate for C. jejuni subunit vaccine.

Keywords: Campylobacter jejuni; Prokaryotic expression; Immunogenicity