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Binding of enterotoxigenic Escherichia coli to isolated enterocytes and intestinal mucus.

Journal article
Authors A Helander
Gunnar C. Hansson
Ann-Mari Svennerholm
Published in Microbial pathogenesis
Volume 23
Issue 6
Pages 335-46
ISSN 0882-4010
Publication year 1997
Published at Institute of Medical Biochemistry
Institute of Medical Microbiology/Immunology
Pages 335-46
Language en
Links dx.doi.org/10.1006/mpat.1997.0163
Keywords Animals, Antigens, Bacterial, metabolism, Antigens, Surface, metabolism, Bacterial Outer Membrane Proteins, metabolism, Bacterial Proteins, metabolism, Cells, Cultured, Endopeptidase K, pharmacology, Escherichia coli, drug effects, physiology, Escherichia coli Proteins, Female, Humans, Intestinal Mucosa, metabolism, microbiology, Intestine, Small, cytology, microbiology, Male, Microscopy, Electron, Microscopy, Immunoelectron, Mitogens, pharmacology, Mucus, drug effects, microbiology, Periodic Acid, pharmacology, Rabbits
Subject categories Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)

Abstract

Binding of human enterotoxigenic Escherichia coli (ETEC) to the small intestine is a prerequisite for colonization and is mediated by colonization factor (CF) antigens. Coli surface antigen 6 (CS6) is considered a CF but binding to isolated enterocytes has not been established. In this study bacteria expressing CS6 were analysed for binding to enterocytes from human and rabbit small intestine, isolated using either an EDTA-containing buffer or a buffer devoid of EDTA. We found that the bacteria bound to enterocytes from rabbit ileum and human duodenum, but only when the cells had been isolated in the absence of EDTA. Pretreatment of rabbit enterocytes with meta-periodate resulted in a decreased proportion of cells with bound bacteria. Purified CS6, and for comparison other ETEC CFs, were also tested for binding to different human and rabbit mucus fractions. These analyses showed that purified CS6 bound to mucus from rabbit duodenum and ileum as well as from human duodenum, jejunum and ileum and that this binding was abolished by pretreatment of the mucus material with meta-periodate or Proteinase K. CFA/I, CS1 to CS5, CS7, CS17, putative CF (PCF) O159 (CS12), PCFO166 (CS14), and CFA/III (CS8) also bound to the rabbit mucus material although with different patterns; the binding of CS2 and CS5 was abolished by meta-periodate treatment. Thus, ETEC bacteria expressing CS6 might bind to carbohydrate-containing structure(s) in the apical membrane of isolated rabbit ileal and human duodenal enterocytes that could probably be released by EDTA treatment. In addition, CS6 and other ETEC CFs bind to component(s), in some instances protein-associated carbohydrate structures, in mucus fractions from small intestine.

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