Role of flagella in adherence, internalization, and translocation of Campylobacter jejuni in nonpolarized and polarized epithelial cell cultures.
Previous studies of Campylobacter jejuni have suggested that flagellin is an adhesin for epithelial cells and that motility is a virulence factor of this bacterium. The role of flagella in the interactions of C. jejuni with nonpolarized and polarized epithelial cells was examined with flagellar mutants. Flagellated, nonmotile (flaA flaB+ Mot-) and nonflagellated, nonmotile (flaA flaB Mot-) mutants of C. jejuni were constructed by in vivo homologous recombination and gene replacement techniques. Both classes of mutants were found to adhere to cells of human epithelial origin (INT 407) equally well; however, on the basis of the percentage of the inoculum internalized, internalization of the flaA flaB Mot- mutants was decreased by factors ranging from approximately 30 to 40 compared with the parent. The flaA flaB+ Mot- mutant was internalized by the INT 407 cells at levels six- to sevenfold higher than the flaA flaB Mot- mutants. Both classes of mutants, unlike the parent, were unable to translocate across polarized Caco-2 monolayers. These results indicate that flagella are not involved in C. jejuni adherence to epithelial cells but that they do play a role in internalization. Furthermore, the results suggest that either the motility of C. jejuni or the product of flaA is essential for the bacterium to cross polarized epithelial cell monolayers.
Grant
C C
CC
Department of Microbiology and Immunology, Stanford University Medical Center, California 94305.
Konkel
M E
ME
Cieplak
W
W
Tompkins
L S
LS
D001422:Bacterial Adhesion; D001483:Base Sequence; D016123:Campylobacter jejuni / Q000235:genetics / Q000472:pathogenicity*; D016764:Cell Polarity; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004252:DNA Mutational Analysis; D004269:DNA, Bacterial / Q000235:genetics; D004705:Endocytosis; D004848:Epithelium / Q000382:microbiology; D005408:Flagellin / Q000235:genetics* / Q000378:metabolism; D066298:In Vitro Techniques; D007422:Intestines / Q000382:microbiology*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D015183:Restriction Mapping
D004269:DNA, Bacterial; D009838:Oligodeoxyribonucleotides; D005408:Flagellin