Ca(2+)/calmodulin-dependent invasion of microvascular endothelial cells of human brain by Escherichia coli K1.
Escherichia coli K1 invasion of microvascular endothelial cells of human brain (HBMEC) is required for E. coli penetration into the central nervous system, but the microbial-host interactions that are involved in this invasion of HBMEC remain incompletely understood. We have previously shown that FimH, one of the E. coli determinants contributing to the binding to and invasion of HBMEC, induces Ca(2+) changes in HBMEC. In the present study, we have investigated in detail the role of cellular calcium signaling in the E. coli K1 invasion of HBMEC, the main constituents of the blood-brain barrier. Addition of the meningitis-causing E. coli K1 strain RS218 (O18:K1) to HBMEC results in transient increases of intracellular free Ca(2+). Inhibition of phospholipase C with U-73122 and the chelating of intracellular Ca(2+) by BAPTA/AM reduces bacterial invasion of HBMEC by approximately 50%. Blocking of transmembrane Ca(2+) fluxes by extracellular lanthanum ions also inhibits the E. coli invasion of HBMEC by approximately 50%. In addition, E. coli K1 invasion is significantly inhibited when HBMEC are pretreated by the calmodulin antagonists, trifluoperazine or calmidazolium, or by ML-7, a specific inhibitor of Ca(2+)/calmodulin-dependent myosin light-chain kinase. These findings indicate that host intracellular Ca(2+) signaling contributes in part to E. coli K1 invasion of HBMEC.
Kim
Yuri V
YV
Division of Pediatrics Infectious Diseases, Johns Hopkins University School of Medicine, Baltimore, MD 21287, USA.
Pearce
Donna
D
Kim
Kwang Sik
KS
D001921:Brain / Q000098:blood supply* / Q000382:microbiology*; D002118:Calcium / Q000378:metabolism*; D020013:Calcium Signaling; D002147:Calmodulin / Q000378:metabolism*; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000378:metabolism / Q000382:microbiology; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism / Q000382:microbiology*; D004926:Escherichia coli / Q000472:pathogenicity*; D006801:Humans; D008833:Microcirculation / Q000166:cytology
D002147:Calmodulin; D002118:Calcium