The lpf gene cluster for long polar fimbriae is not involved in adherence of enteropathogenic Escherichia coli or virulence of Citrobacter rodentium.
Using the enteropathogenic Escherichia coli (EPEC) genome sequence, we found that EPEC E2348/69 has an lpfABCDE gene cluster homologous (about 60% identical at the protein level) to the Salmonella long polar fimbria (LPF) operon. To determine whether this operon is essential for adherence, the lpfABCD(E2)(3) genes were deleted from EPEC strain E2348/69 by allelic exchange. Analysis of the resulting EPECDeltalpfABCD(E23) strain showed no change in adherence to HeLa cells or to human intestinal biopsy cells in the in vitro organ culture (IVOC) system compared to the wild type. Sera from volunteers experimentally infected with E2348/69 showed no antibody response to the major subunit protein, LpfA. These results suggested that the lpf(E23) gene cluster is not necessary for EPEC adherence and attaching/effacing (A/E) lesion formation on human biopsy samples and is not expressed during human infection. We also identified an lpf gene cluster in Citrobacter rodentium strain ICC168 (lpf(cr)). A DeltalpfA(cr) mutant of ICC168 retained wild-type adherence and A/E lesion-forming activity on HeLa cells. C3H/HeJ mice were infected with a wild-type C. rodentium strain and its lpfA(cr) isogenic mutant. Both strains were recovered at high levels in stools, and there were no significant differences between the groups both in terms of the number of CFU/organ (colon and cecum) and in terms of the amount of hyperplasia, as measured by weight. Similar results were observed in a second mouse strain, C57BL/6. These data suggest that in addition to playing no apparent role in EPEC pathogenesis, lpf(cr) is not required for C. rodentium virulence in either the C3H/HeJ or C57BL/6 mouse model.
Tatsuno
Ichiro
I
Center for Vaccine Development, University of Maryland School of Medicine, 685 West Baltimore Street, Baltimore, MD 21201, USA.
Mundy
Rosanna
R
Frankel
Gad
G
Chong
Yuwen
Y
Phillips
Alan D
AD
Torres
Alfredo G
AG
Kaper
James B
JB
D000818:Animals; D001422:Bacterial Adhesion / Q000235:genetics*; D044082:Citrobacter rodentium / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity*; D004195:Disease Models, Animal; D004756:Enterobacteriaceae Infections / Q000382:microbiology* / Q000473:pathology; D004927:Escherichia coli Infections / Q000382:microbiology / Q000473:pathology; D019453:Escherichia coli O157 / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D010861:Fimbriae, Bacterial / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D005798:Genes, Bacterial; D006367:HeLa Cells; D006801:Humans; D007421:Intestine, Small / Q000382:microbiology / Q000473:pathology; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D005810:Multigene Family; D009154:Mutation; D009924:Organ Culture Techniques