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


CONCLUSIONS
rPEB1 protein is a valuable candidate for C. jejuni subunit vaccine.
Du
Lian-Feng
LF
Department of Immunology, Zhuhai Campus of Zunyi Medical College, Zhuhai 519041, Guangdong Province, China.
Li
Zhen-Jiang
ZJ
Tang
Xian-Ying
XY
Huang
Jun-Qiong
JQ
Sun
Wan-Bang
WB
D000284:Administration, Oral; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000942:Antigens, Bacterial / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D001428:Bacterial Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D002169:Campylobacter Infections / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control*; D016123:Campylobacter jejuni / Q000235:genetics / Q000276:immunology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004195:Disease Models, Animal; D004306:Dose-Response Relationship, Immunologic; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors; D007273:Injections, Intramuscular; D007279:Injections, Subcutaneous; D008213:Lymphocyte Activation; D051379:Mice; D008807:Mice, Inbred BALB C; D013601:T-Lymphocytes / Q000276:immunology; D013997:Time Factors; D014614:Vaccines, Synthetic / Q000276:immunology
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001428:Bacterial Vaccines; C489977:PEB1 protein, Campylobacter; D014614:Vaccines, Synthetic