title: In vitro binding of Campylobacter jejuni/coli outer membrane preparations to INT 407 cell membranes.
abstract: Outer membrane fractions (OMs) of nine Campylobacter (C.) jejuni and two C. coli strains belonging to different serovars, from human and various animal origins, were extracted by treatment with sodium N-lauryl sarcosinate. Using n-octyl-beta-D-glucopyranoside a 42-kDa protein and a flagella-enriched fraction were obtained. The capacity of the crude bacterial OM preparations, the purified 42-kDa protein and the flagella to bind to membranes of the human embryonic intestinal cell line INT 407 was tested by an enzyme-linked immunosorbent assay. The crude OM and the 42-kDa-enriched fraction were found to bind very well to the cell membranes, whereas the flagella preparation showed only a weak binding. Using monoclonal antibodies (mAbs) with HS 2-lipopolysaccharide (LPS) specificity, binding of crude HS 2 strain OM preparations to cell membranes was detected in a significant range, whereas with flagellin-specific mAbs binding of OMs and flagella to cell membranes was only detected to a very low extent. Binding of OMs to cell membranes was inhibited by preincubation of OMs with serovar-specific mouse hyperimmune serum, whereas on preincubation with mAbs directed against LPS or flagella binding was practically not inhibited. OMs extracted after pretreatment of the bacteria with proteinase K showed an altered SDS-PAGE pattern especially for the 42-kDa protein subunit and and their capacity to bind to cell membranes was significantly reduced. The binding was also reduced by preincubation of the OMs with L-fucose or D-mannose.
authors.affiliation: Institut für Mikrobiologie und Tierhygiene, Fachbereich Veterinärmedizin, Berlin, Federal Republic of Germany.
authors.forename: I
authors.initials: I
authors.lastname: Moser
authors.forename: W F
authors.initials: WF
authors.lastname: Schröder
authors.forename: E
authors.initials: E
authors.lastname: Hellmann
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology / Q000378:metabolism*; D017000:Campylobacter coli / Q000276:immunology / Q000378:metabolism*; D016123:Campylobacter jejuni / Q000276:immunology / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism* / Q000502:physiology; D046508:Culture Techniques; D004591:Electrophoresis, Polyacrylamide Gel; D019286:Endopeptidase K; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005407:Flagella / Q000378:metabolism; D005643:Fucose / Q000494:pharmacology; D006801:Humans; D008070:Lipopolysaccharides / Q000276:immunology; D008358:Mannose / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D012697:Serine Endopeptidases
chemical_list: D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D008070:Lipopolysaccharides; D005643:Fucose; D012697:Serine Endopeptidases; D019286:Endopeptidase K; D008358:Mannose