Immunogenicity and immunoprotection of recombinant PEB1 in Campylobacter-jejuni-infected mice

• Published in: World journal of gastroenterology : WJG 2008-10, Vol.14 (40), p.6244-6248
• Main Authors: Du, Lian-Feng, Li, Zhen-Jiang, Tang, Xian-Ying, Huang, Jun-Qiong, Sun, Wan-Bang
• Format: Article
• Language: English
• Subjects: Administration, Oral; Animals; Antibodies, Bacterial - blood; Antigens, Bacterial - administration & dosage; Antigens, Bacterial - genetics; Antigens, Bacterial - immunology; Bacterial Vaccines - administration & dosage; Bacterial Vaccines - genetics; Bacterial Vaccines - immunology; Campylobacter Infections - immunology; Campylobacter Infections - microbiology; Campylobacter Infections - prevention & control; Campylobacter jejuni - genetics; Campylobacter jejuni - immunology; Cell Proliferation; Cells, Cultured; Cloning, Molecular; Disease Models, Animal; Dose-Response Relationship, Immunologic; Escherichia coli - genetics; Escherichia coli - metabolism; Genetic Vectors; Injections, Intramuscular; Injections, Subcutaneous; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Rapid Communication; T-Lymphocytes - immunology; Time Factors; Vaccines, Synthetic - immunology; 免疫原性; 原核表达
• ISSN: 1007-9327; 2219-2840
• DOI: 10.3748/wjg.14.6244

AIM： To construct a prokaryotic expression vector carrying Campylobacterjejuni peblA 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： peblA gene was amplified by PCR, target gene and prokaryotic expression ptasmid pET28a （＋） was digested with BamHI and XhoI, respectively. DNA was ligated with T4 DNA ligase to construct recombinant plasmid pET28a（＋）-peblA. The rPEB1 was expressed in E. coli BL21 （DE3） and identified by SDS-PAGE. BALB/c mice were immunized with rPEBI. 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 （＋）-peblA was correctly constructed. The expression output of PEB1 protein in pET28a （＋）-peblA system was approximately 33% of total proteins in E. coil 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.