title: Analysis and application of a neutralizing linear epitope on liable toxin B of enterotoxin Escherichia coli.
abstract: Heat-labile enterotoxin (LT) of enterotoxigenic Escherichia coli (ETEC) is one of the major virulence factors for causing diarrhea in piglets, and LT is a strong immunogen. Thus, LT represents an important target for development of vaccines and diagnostic tests. In this study, bioinformatic tools were used to predict six antigenic B cell epitopes in the B subunit of LT protein (LTB) of ETEC strains. Then, seven antigenic B cell epitopes of LTB were identified by polyclonal antisera (polyclonal antibody (PAb)) using a set of LTB-derived peptides expressed as maltose-binding protein (MBP) fusion protein. In addition, one LTB-specific monoclonal antibody (MAb) was generated and defined its corresponding epitope as mentioned above. This MAb was able to specifically bind with native LT toxin and has no cross-reaction with LT-II (type II heat-labile enterotoxin), Stx1 (Shiga toxin I), Stx2 (Shiga toxin II), STa (heat-stable enterotoxin I), and STb (heat-stable enterotoxin II) toxins. Further, this MAb was able to interrupt LT toxin specific binding to GM1 receptor, indicating that the corresponding epitope is the specific binding region to GM1 receptor. Moreover, in vitro and in vivo assay showed that the MAb was able to neutralize the native LT toxin. Diarrheal suckling pigs challenged with LT-positive ETEC strain recovered when an enema with this purified MAb was administered. This study will provide the foundation for further studies about the interaction between LT toxin and GM1 receptor and about the developing of epitope-based vaccines and specific therapeutic agent.
authors: Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, Harbin, Heilongjiang, 150030, People's Republic of China.
authors: Weikun
authors: W
authors: Guan
authors: Wenxin
authors: W
authors: Liu
authors: Jun
authors: J
authors: Bao
authors: Jinping
authors: J
authors: Li
authors: Chaowen
authors: C
authors: Yuan
authors: Jie
authors: J
authors: Tang
authors: Dongfang
authors: D
authors: Shi
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D057134:Antibodies, Neutralizing / Q000276:immunology*; D001427:Bacterial Toxins / Q000235:genetics / Q000276:immunology*; D019295:Computational Biology; D003967:Diarrhea / Q000628:therapy; D004733:Enema; D004768:Enterotoxins / Q000235:genetics / Q000276:immunology*; D018985:Epitopes, B-Lymphocyte / Q000235:genetics / Q000276:immunology*; D004927:Escherichia coli Infections / Q000628:therapy / Q000662:veterinary; D029968:Escherichia coli Proteins / Q000235:genetics / Q000276:immunology*; D007116:Immunization, Passive / Q000379:methods; D009500:Neutralization Tests; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000378:metabolism; D013552:Swine; D013553:Swine Diseases / Q000628:therapy; D016896:Treatment Outcome
chemical_list: D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D001427:Bacterial Toxins; D004768:Enterotoxins; D018985:Epitopes, B-Lymphocyte; D029968:Escherichia coli Proteins; D011956:Receptors, Cell Surface; C034993:ganglioside receptor; C027538:heat-labile enterotoxin, E coli