title: Inhibition of adhesion of Escherichia coli k88ac fimbria to its receptor, intestinal mucin-type glycoproteins, by a monoclonal antibody directed against a variable domain of the fimbria.
abstract: Strains of enterotoxigenic Escherichia coli that express K88 fimbriae are among the most common causes of diarrhea in young pigs. Adhesion of bacteria to receptors on intestinal epithelial cells, mediated by K88 fimbriae, is the initial step in the establishment of infection. Three antigenic variants of K88 fimbriae exist in nature: K88ab, K88ac, and K88ad. K88ac is the most prevalent and may be the only variant of significance in swine disease. Each K88 fimbrial variant is composed of multiple antigenic determinants. Some of these determinants are shared among the three variants and may be referred to as conserved epitopes, whereas others are unique to a specific variant and may be referred to as variable epitopes. In this study, monoclonal antibodies (MAbs) specific to either variable or conserved epitopes of K88ac fimbriae were produced. The specificity of each MAb was tested by enzyme-linked immunosorbent and immunoblot assays. Fab fragments were prepared from these MAbs and were tested for their ability to block the binding of K88-positive bacteria and purified fimbriae to porcine enterocyte brush border vesicles and purified K88 receptors, respectively. The purified receptors were intestinal mucin-type sialoglycoproteins (IMTGP) isolated from porcine enterocytes (A. K. Erickson, D. R. Baker, B. T. Bosworth, T. A. Casey, D. A. Benfield, and D. H. Francis, Infect. Immun. 62:5404-5410, 1994). Fab fragments prepared from MAbs specific for variable epitopes blocked the binding of bacteria to brush borders and of fimbriae to IMTGP. However, those from MAbs specific for a conserved epitope did not. These observations indicate that the receptor-binding domain of a K88ac fimbria is contained, at least in part, within the antigenically variable epitopes of that fimbria. Epitope mapping for one of the MAbs, which recognizes a linear epitope on K88ac fimbriae, indicated that this MAb binds to the region from amino acid no. 64 to no. 107 on the major subunit of K88ac fimbriae.
authors.affiliation: Department of Veterinary Science, South Dakota State University, Brookings, South Dakota 57007-1396, USA.
authors.forename: R
authors.initials: R
authors.lastname: Sun
authors.forename: T J
authors.initials: TJ
authors.lastname: Anderson
authors.forename: A K
authors.initials: AK
authors.lastname: Erickson
authors.forename: E A
authors.initials: EA
authors.lastname: Nelson
authors.forename: D H
authors.initials: DH
authors.lastname: Francis
mesh_terms: D018830:Adhesins, Escherichia coli / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D000954:Antigens, Surface / Q000276:immunology / Q000378:metabolism*; D001422:Bacterial Adhesion; D020895:Enterocytes / Q000378:metabolism; D018604:Epitope Mapping; D004926:Escherichia coli / Q000472:pathogenicity*; D029968:Escherichia coli Proteins; D036781:Fimbriae Proteins; D010861:Fimbriae, Bacterial / Q000276:immunology / Q000378:metabolism*; D007413:Intestinal Mucosa / Q000166:cytology; D008871:Microvilli / Q000378:metabolism; D009077:Mucins / Q000276:immunology / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000276:immunology / Q000378:metabolism*; D013552:Swine
chemical_list: D018830:Adhesins, Escherichia coli; D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D000954:Antigens, Surface; D029968:Escherichia coli Proteins; C026888:K88 antigen, E coli; D009077:Mucins; D011956:Receptors, Cell Surface; D036781:Fimbriae Proteins