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Flagella Promote Escherichia coli K1 Association with and Invasion of Human Brain Microvascular Endothelial Cells

Author:
Parthasarathy, G.   Yao, Y.   Kim, K. S.  


Journal:
Infection and Immunity


Issue Date:
2007


Abstract(summary):

Escherichia coli containing the K1 capsule is the leading cause of gram-negative meningitis, but the pathogenesis of this disease is not completely understood. Recent microarray experiments in which we compared the gene expression profile of E. coli K1 associated with human brain microvascular endothelial cells (HBMEC) to the gene expression profile of E. coli KI not associated with HBMEC revealed that there was a threefold increase in the expression of the fliI gene, encoding an ATP synthase involved in flagellar synthesis and motility, in HBMEC-associated E. coli. In this study, we examined the role of flagella in E. coli KI association with and invasion of HBMEC by constructing isogenic Delta flhDC, Delta fliI, Delta fliC, and Delta cheW mutants that represented each class of flagellar genes. Mutations that affected the flagellum structure and flagellum formation (Delta flhDC, Delta fliI, and Delta fliC) resulted in significant defects in motility, as well as in HBMEC association and invasion, compared to the characteristics of the wild-type strain when preparations were examined with or without centrifugation. Transcomplementation with the corresponding genes restored the levels of these mutants to the levels of the parent strain. These findings suggest that the HBMEC association and invasion defects of the mutants are most likely related to flagella and less likely due to their motility defects. This conclusion was supported by our demonstration that the cheW mutant was not motile but was able to associate with and invade HBMEC. In addition, purified recombinant flagellin reduced the association of the wild-type strain with HBMEC by similar to 40%, while it had no effect on the fliC mutant's association with HBMEC. Together, these findings indicate that flagella promote E. coli KI binding to HBMEC.


Page:
2937-2945


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