The immunogenicity of fusion protein linking the carboxyl terminus of the B subunit of Shiga toxin 2 to the B subunit of E. coli heat-labile enterotoxin.
To augment the immunogenicity of the subunit B of Shiga toxin (Stx2e B) produced by Escherichia coli and protect piglets from edema disease in china, a fusion gene was constructed consisting of Stx2e B genetically linked at the N-terminus of the B subunit of heat-labile enterotoxin (LTB) in a translational fusion. After being induced with IPTG, the expressed fusion protein of Stx2e B-LTB was about 8.8% of total proteins, approximately 13 microg/ml of the bacteria culture. The Stx2e B-LTB fusion protein was found to be nontoxic to Vero cells at the dose higher than 1 microg/ml and to mice less than 100 microg/ml. Antibody titer against the fusion protein Stx2e B-LTB was 1:76,800, much higher than that of the recombinant Stx2e B protein (1:12,800) alone. All of the mice immunized with the Stx2e B-LTB fusion protein survived when challenged with a lethal dose (LD) of Stx2e toxin. The results showed that the poor immunogenicity of Stx2e B was overcome by conjugating the stx2e B to ltB. The immunogenicity of the constructed fusion protein Stx2e B-LTB in the present study was highly qualified to protect animals against Shiga toxin produced from Shiga toxin-producing Escherichia coli (STEC). The fusion protein of Stx2e B-LTB could be a candidate for a vaccine against edema disease and post-weaning diarrhea simultaneously in piglets.
Ran
Xue Qin
XQ
Faculty of Animal Science and Veterinary Medicine, Guizhou University, Xueshi Road, Guiyang, Guizhou Province, China.
Wang
Hong Zhen
HZ
Liu
Jin Juan
JJ
Li
Sheng
S
Wang
Jia Fu
JF
D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000378:metabolism; D001427:Bacterial Toxins / Q000235:genetics / Q000276:immunology*; D001428:Bacterial Vaccines / Q000276:immunology*; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D004768:Enterotoxins / Q000235:genetics / Q000276:immunology*; D004926:Escherichia coli / Q000276:immunology; D004927:Escherichia coli Infections / Q000517:prevention & control*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000276:immunology*; D051379:Mice; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology* / Q000633:toxicity; D022641:Shiga Toxin 2 / Q000235:genetics / Q000276:immunology*; D016019:Survival Analysis; D013552:Swine; D013553:Swine Diseases / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control; D014614:Vaccines, Synthetic / Q000276:immunology; D014709:Vero Cells
D000907:Antibodies, Bacterial; D001427:Bacterial Toxins; D001428:Bacterial Vaccines; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; D022641:Shiga Toxin 2; D014614:Vaccines, Synthetic; C450841:shiga toxin 2 B subunit; C027538:heat-labile enterotoxin, E coli