Abstract
Strains of enterotoxigenic Escherichia coli that produce 987P fimbriae (987P+ strains) colonize the small intestines and cause diarrhea in neonatal (< 6-day-old) pigs but not in weaned pigs. However, 987P+ E. coli strains adhere in vitro to intestinal epithelial cells from pigs of both ages. Two intestinal components, designated 987R and 987M, bind 987P fimbriae (987P) on Western blots (immunoblots). We examined brush borders (BB) and intestinal washes (IW) from pigs to determine if they contain glycolipids which bind 987P. Total glycolipid extracts from BB and IW of 4-week-old pigs were separated on thin-layer chromatograms and overlaid with purified 987P. Bound 987P were detected with 987P-specific antiserum. 987P bound to at least one moiety in both BB and IW glycolipids and also bound to several purified glycolipids, including gangliotetraosylceramide, lactosylceramide (CDH), sulfatide (SFT), gangliotriaosylceramide, and galactosylceramide (listed in order of decreasing affinity). Strain 987, but not the isogenic 987P- strain I36, bound to these same glycolipids, indicating that the fimbriae contain the adhesin required for binding to these lipids. Glycolipids extracted from BB and IW isolated from 3- and 4-week-old pigs and from BB isolated from 1-day-old pigs contained similar amounts of glycolipids like CDH and SFT that bound 987P. Finally, 987P bound to CDH, SFT, and total BB glycolipids separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to Immobilon, and these glycolipids had mobilities similar to that of 987M. Thus, 987M may contain 987P-binding glycolipids. We hypothesize that glycolipid receptors for 987P, most likely CDH or SFT, in the mucus of older pigs bind 987P and inhibit 987P- mediated intestinal colonization by preventing the attachment of 987P+ E. coli to 987P receptors on the intestinal epithelium.
Full text
PDF





Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Dean E. A. Comparison of receptors for 987P pili of enterotoxigenic Escherichia coli in the small intestines of neonatal and older pig. Infect Immun. 1990 Dec;58(12):4030–4035. doi: 10.1128/iai.58.12.4030-4035.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dean E. A., Isaacson R. E. In vitro adhesion of piliated Escherichia coli to small intestinal villous epithelial cells from rabbits and the identification of a soluble 987P pilus receptor-containing fraction. Infect Immun. 1982 Jun;36(3):1192–1198. doi: 10.1128/iai.36.3.1192-1198.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dean E. A., Isaacson R. E. Purification and characterization of a receptor for the 987P pilus of Escherichia coli. Infect Immun. 1985 Jan;47(1):98–105. doi: 10.1128/iai.47.1.98-105.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dean E. A., Whipp S. C., Moon H. W. Age-specific colonization of porcine intestinal epithelium by 987P-piliated enterotoxigenic Escherichia coli. Infect Immun. 1989 Jan;57(1):82–87. doi: 10.1128/iai.57.1.82-87.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Erickson A. K., Willgohs J. A., McFarland S. Y., Benfield D. A., Francis D. H. Identification of two porcine brush border glycoproteins that bind the K88ac adhesin of Escherichia coli and correlation of these glycoproteins with the adhesive phenotype. Infect Immun. 1992 Mar;60(3):983–988. doi: 10.1128/iai.60.3.983-988.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Isaacson R. E., Richter P. Escherichia coli 987P pilus: purification and partial characterization. J Bacteriol. 1981 May;146(2):784–789. doi: 10.1128/jb.146.2.784-789.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kyogashima M., Ginsburg V., Krivan H. C. Escherichia coli K99 binds to N-glycolylsialoparagloboside and N-glycolyl-GM3 found in piglet small intestine. Arch Biochem Biophys. 1989 Apr;270(1):391–397. doi: 10.1016/0003-9861(89)90042-8. [DOI] [PubMed] [Google Scholar]
- Morris J. A., Sojka W. J., Wells G. A. K99 and 987P adhesins on Escherichia coli enteropathogenic for piglets. Vet Rec. 1982 Aug 21;111(8):165–166. doi: 10.1136/vr.111.8.165. [DOI] [PubMed] [Google Scholar]
- Nagy B., Casey T. A., Whipp S. C., Moon H. W. Susceptibility of porcine intestine to pilus-mediated adhesion by some isolates of piliated enterotoxigenic Escherichia coli increases with age. Infect Immun. 1992 Apr;60(4):1285–1294. doi: 10.1128/iai.60.4.1285-1294.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nagy B., Moon H. W., Isaacson R. E. Colonization of porcine intestine by enterotoxigenic Escherichia coli: selection of piliated forms in vivo, adhesion of piliated forms to epithelial cells in vitro, and incidence of a pilus antigen among porcine enteropathogenic E. coli. Infect Immun. 1977 Apr;16(1):344–352. doi: 10.1128/iai.16.1.344-352.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Payne D., O'Reilly M., Williamson D. The K88 fimbrial adhesin of enterotoxigenic Escherichia coli binds to beta 1-linked galactosyl residues in glycosphingolipids. Infect Immun. 1993 Sep;61(9):3673–3677. doi: 10.1128/iai.61.9.3673-3677.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Peterson G. L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346–356. doi: 10.1016/0003-2697(77)90043-4. [DOI] [PubMed] [Google Scholar]
- Runnels P. L., Moon H. W., Schneider R. A. Development of resistance with host age to adhesion of K99+ Escherichia coli to isolated intestinal epithelial cells. Infect Immun. 1980 Apr;28(1):298–300. doi: 10.1128/iai.28.1.298-300.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Samuel J. E., Perera L. P., Ward S., O'Brien A. D., Ginsburg V., Krivan H. C. Comparison of the glycolipid receptor specificities of Shiga-like toxin type II and Shiga-like toxin type II variants. Infect Immun. 1990 Mar;58(3):611–618. doi: 10.1128/iai.58.3.611-618.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sarmiento J. I., Casey T. A., Moon H. W. Postweaning diarrhea in swine: experimental model of enterotoxigenic Escherichia coli infection. Am J Vet Res. 1988 Jul;49(7):1154–1159. [PubMed] [Google Scholar]
- Seignole D., Mouricout M., Duval-Iflah Y., Quintard B., Julien R. Adhesion of K99 fimbriated Escherichia coli to pig intestinal epithelium: correlation of adhesive and non-adhesive phenotypes with the sialoglycolipid content. J Gen Microbiol. 1991 Jul;137(7):1591–1601. doi: 10.1099/00221287-137-7-1591. [DOI] [PubMed] [Google Scholar]
- Sellwood R., Gibbons R. A., Jones G. W., Rutter J. M. Adhesion of enteropathogenic Escherichia coli to pig intestinal brush borders: the existence of two pig phenotypes. J Med Microbiol. 1975 Aug;8(3):405–411. doi: 10.1099/00222615-8-3-405. [DOI] [PubMed] [Google Scholar]
- Sherman P. M., Boedeker E. C. Pilus-mediated interactions of the Escherichia coli strain RDEC-1 with mucosal glycoproteins in the small intestine of rabbits. Gastroenterology. 1987 Oct;93(4):734–743. doi: 10.1016/0016-5085(87)90435-5. [DOI] [PubMed] [Google Scholar]
- Teneberg S., Willemsen P., de Graaf F. K., Karlsson K. A. Receptor-active glycolipids of epithelial cells of the small intestine of young and adult pigs in relation to susceptibility to infection with Escherichia coli K99. FEBS Lett. 1990 Apr 9;263(1):10–14. doi: 10.1016/0014-5793(90)80693-d. [DOI] [PubMed] [Google Scholar]
- Wilson R. A., Francis D. H. Fimbriae and enterotoxins associated with Escherichia coli serogroups isolated from pigs with colibacillosis. Am J Vet Res. 1986 Feb;47(2):213–217. [PubMed] [Google Scholar]
- Yang Z., Bergström J., Karlsson K. A. Glycoproteins with Gal alpha 4Gal are absent from human erythrocyte membranes, indicating that glycolipids are the sole carriers of blood group P activities. J Biol Chem. 1994 May 20;269(20):14620–14624. [PubMed] [Google Scholar]