title: Heat-labile- and heat-stable-toxoid fusions (LTR₁₉₂G-STaP₁₃F) of human enterotoxigenic Escherichia coli elicit neutralizing antitoxin antibodies.
abstract: Enterotoxigenic Escherichia coli (ETEC) strains are a major cause of diarrheal disease in humans and animals. Adhesins and enterotoxins, including heat-labile (LT) and heat-stable (STa) toxins, are the key virulence factors. Antigenic adhesin and LT antigens have been used in developing vaccines against ETEC diarrhea. However, STa has not been included because of its poor immunogenicity and potent toxicity. Our recent study showed that porcine-type STa toxoids became immunogenic and elicited neutralizing anti-STa antibodies after being genetically fused to a full-length porcine-type LT toxoid, LT(R₁₉₂G) (W. Zhang et al., Infect. Immun. 78:316-325, 2010). In this study, we mutated human-type LT and STa genes, which are highly homologous to porcine-type toxin genes, for a full-length LT toxoid (LT(R₁₉₂)) and a full-length STa toxoid (STa(P₁₃F)) and genetically fused them to produce LT₁₉₂-STa₁₃ toxoid fusions. Mice immunized with LT₁₉₂-STa₁₃ fusion antigens developed anti-LT and anti-STa IgG (in serum and feces) and IgA antibodies (in feces). Moreover, secretory IgA antibodies from immunized mice were shown to neutralize STa and cholera toxins in T-84 cells. In addition, we fused the STa₁₃ toxoid at the N terminus and C terminus, between the A1 and A2 peptides, and between the A and B subunits of LT₁₉₂ to obtain different fusions in order to explore strategies for enhancing STa immunogenicity. This study demonstrated that human-type LT₁₉₂-STa₁₃ fusions induce neutralizing antitoxin antibodies and provided important information for developing toxoid vaccines against human ETEC diarrhea.
authors.affiliation: Veterinary & Biomedical Sciences Department/The Center for Infectious Disease Research & Vaccinology, South Dakota State University, Brookings, SD 57006, USA.
authors.forename: Mei
authors.initials: M
authors.lastname: Liu
authors.forename: Xiaosai
authors.initials: X
authors.lastname: Ruan
authors.forename: Chengxian
authors.initials: C
authors.lastname: Zhang
authors.forename: Steve R
authors.initials: SR
authors.lastname: Lawson
authors.forename: David E
authors.initials: DE
authors.lastname: Knudsen
authors.forename: James P
authors.initials: JP
authors.lastname: Nataro
authors.forename: Donald C
authors.initials: DC
authors.lastname: Robertson
authors.forename: Weiping
authors.initials: W
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood*; D057134:Antibodies, Neutralizing / Q000097:blood*; D001427:Bacterial Toxins / Q000235:genetics* / Q000276:immunology; D054307:Enterotoxigenic Escherichia coli / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D004768:Enterotoxins / Q000235:genetics* / Q000276:immunology; D004927:Escherichia coli Infections / Q000517:prevention & control; D029968:Escherichia coli Proteins / Q000235:genetics* / Q000276:immunology; D005260:Female; D005818:Genetic Engineering; D006801:Humans; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000276:immunology; D014121:Toxoids
chemical_list: D000907:Antibodies, Bacterial; D057134:Antibodies, Neutralizing; D001427:Bacterial Toxins; D004768:Enterotoxins; D029968:Escherichia coli Proteins; D011993:Recombinant Fusion Proteins; D014121:Toxoids; C034151:heat stable toxin (E coli); C027538:heat-labile enterotoxin, E coli