Adhesion of enterotoxigenic (ETEC) and bovine mastitis Escherichia coli strains to rat embryonic fibroblasts: role of amino-terminal domain of fibronectin in bacterial adhesion.
Adherence of human enterotoxigenic and bovine mastitis Escherichia coli to rat embryonic fibroblasts was studied. Adhesion of E. coli strains B34289c (human) and 1407 (bovine) was rapid and reached maximum after 30-40 min. Strain 1410 (bovine), which binds fibronectin but not its 29K amino-terminal fragment, did not adhere to the fibroblasts. Strain B34289c grown at 25 C or below and at 40 C or above lost its binding and adhesive properties simultaneously. Maximum binding and adhesion for this strain was achieved when it was grown at 33 C. Strains grown at this temperature adsorbed to fibronectin-, 29K fragment-, and Octyl Sepharose, with the exception of bovine strain 1410, which did not adsorb to 29K-Sepharose as expected. None of the strains adsorbed to cross-linked Sepharose 4B. 29K-IgG and Fab fragments thereof specifically blocked both binding (max 55%) and adhesion (greater than 95%). Sonicated and trypsin-treated bacteria were no longer able to bind or adhere. The supernatant of sonicated bacteria inhibited both binding and adhesion. Penicillin G at 0.5 micrograms/ml (1/5 minimal inhibitory concentration: MIC) and tetracycline at 0.2 micrograms/ml (1/5 MIC), when included in the growth medium, suppressed the cell surface components responsible for fibronectin binding and fibroblast adhesion. The presence of fibronectin was demonstrated in the fibroblast extracellular matrix by immunofluorescens with 29K-IgG antibodies.
Faris
A
A
Connective Tissue Laboratory, Diabetes Hospital, University of Alabama, Birmingham 35294.
Fröman
G
G
Switalski
L
L
Höök
M
M
D000818:Animals; D000906:Antibodies / Q000276:immunology; D001422:Bacterial Adhesion; D002417:Cattle; D002478:Cells, Cultured; D004926:Escherichia coli / Q000502:physiology*; D005347:Fibroblasts / Q000166:cytology; D005353:Fibronectins / Q000276:immunology / Q000502:physiology*; D066298:In Vitro Techniques; D007700:Kinetics; D008414:Mastitis, Bovine / Q000382:microbiology; D008970:Molecular Weight; D051381:Rats; D013329:Structure-Activity Relationship; D013696:Temperature
D000906:Antibodies; D005353:Fibronectins