title: Bacterial invasion and transcytosis in transfected human brain microvascular endothelial cells.
abstract: Most cases of neonatal bacterial meningitis develop as a result of a hematogenous spread, but it is not clear how circulating bacteria cross the blood-brain barrier. Attempts to answer these questions have been hampered by the lack of a reliable model of the human blood-brain barrier. Human brain microvascular endothelial cells (HBMEC) were isolated and transfected with a pBR322 based plasmid containing simian virus 40 large T antigen (SV40-LT). The transfected HBMEC exhibited similar brain endothelial cell characteristics as the primary HBMEC, i.e. gamma glutamyl transpeptidase and a high transendothelial electrical resistance. Escherischia coli and Citrobacter spp, two important Gram-negative bacilli causing neonatal meningitis, were found to transcytose across primary and transfected HBMEC, without affecting the integrity of the monolayer. In addition, E. coli and C. freundii invaded transfected HBMEC as shown previously with primary HBMEC. We conclude that E. coli and C. freundii are able to invade and transcytose HBMEC and these bacterial-HBMEC interactions are similar between primary and transfected HBMEC. Therefore, our transfected HBMEC should be useful for studying pathogenesis of CNS infections.
authors.affiliation: Division of Infectious Diseases, Children's Hospital Los Angeles, Los Angeles, CA, USA.
authors.forename: M F
authors.initials: MF
authors.lastname: Stins
authors.forename: J
authors.initials: J
authors.lastname: Badger
authors.forename: K
authors.initials: K
authors.lastname: Sik Kim
mesh_terms: D001812:Blood-Brain Barrier; D001921:Brain / Q000098:blood supply*; D002478:Cells, Cultured; D016971:Citrobacter freundii / Q000472:pathogenicity* / Q000502:physiology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000382:microbiology*; D004756:Enterobacteriaceae Infections / Q000382:microbiology; D004926:Escherichia coli / Q000472:pathogenicity* / Q000502:physiology*; D004927:Escherichia coli Infections / Q000382:microbiology; D006801:Humans; D008833:Microcirculation; D014162:Transfection