Adhesion of colonization factor antigen II-positive enterotoxigenic Escherichia coli strains to human enterocytelike differentiated HT-29 cells: a basis for host-pathogen interactions in the gut.
Enterotoxigenic Escherichia coli are the most common cause of travelers' and infant diarrhea in less-developed countries. In the present work, among several metabolically labeled human diarrheagenic E. coli strains, enterotoxigenic strains expressing colonization factor antigen II were shown to bind to HT-29 intestinal cell monolayers when these cells were grown in conditions promoting their enterocytic differentiation. Indirect immunofluorescence with fimbrial antisera revealed that pathogen attachment was associated with the production of a specific bacterial adhesin, the E. coli surface antigen CS3. Scanning and transmission electron micrographs showed an apical pattern of colonization, characteristic of enterotoxigenic E. coli infections. The above data were consistent with all observations previously made with human enterocytes obtained from intestinal biopsies. The lectin-carbohydrate nature of this cell-cell recognition mechanism was also established. Bacterial binding to differentiated HT-29 cells was inhibited by a mixture of newborn meconium glycopeptides. By coating the cell layers with the plant agglutinin from Evonymus europaea, pathogen attachment was also prevented. Binding of 125I-labeled CS3 adhesin and E. europaea agglutinin to brush border membrane proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to nitrocellulose revealed three bands of about 30, 20, and 13 kilodaltons, which acted as receptors for both bacterial and plant lectins. These data suggest that the sugar units to which the bacterial colonization factor CS3 binds are synthesized as carbohydrate chains of three brush border membrane glycoproteins in HT-29 cells by a differentiation-specific pathway.
Neeser
J R
JR
Nestlé Research Centre, Nestec Limited, Lausanne, Switzerland.
Chambaz
A
A
Golliard
M
M
Link-Amster
H
H
Fryder
V
V
Kolodziejczyk
E
E
D000942:Antigens, Bacterial; D001422:Bacterial Adhesion; D003106:Colon / Q000166:cytology / Q000382:microbiology*; D003967:Diarrhea / Q000382:microbiology*; D004768:Enterotoxins / Q000096:biosynthesis; D004926:Escherichia coli / Q000166:cytology*; D036781:Fimbriae Proteins; D005455:Fluorescent Antibody Technique; D006801:Humans; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000382:microbiology; D008470:Meconium / Q000378:metabolism / Q000382:microbiology; D008562:Membrane Glycoproteins / Q000378:metabolism; D008854:Microscopy, Electron; D008871:Microvilli / Q000382:microbiology; D008970:Molecular Weight; D011971:Receptors, Immunologic / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D000942:Antigens, Bacterial; D004768:Enterotoxins; D008562:Membrane Glycoproteins; D011971:Receptors, Immunologic; C047105:bacterial adhesin receptor; C022924:colonization factor antigens; D036781:Fimbriae Proteins