Escherichia coli invasion of brain microvascular endothelial cells in vitro and in vivo: Molecular cloning and characterization of invasion gene ibe10

S. H. Huang, C. Wass, Q. Fu, N. V. Prasadarao, Monique Stins, Kwang Sik Kim

Research output: Contribution to journalArticle

Abstract

Most cases of neonatal Escherichia coli meningitis develop as a result of hematogenous spread, but it is not clear how circulating E. coli crosses the blood-brain barrier. In an attempt to identify E. coli structures contributing to invasion into the central nervous system (CNS), TnphoA mutagenesis was performed with an invasive CSF isolate of E. coli K1 strain RS218 (O18:K1:H7), and TnphoA mutants were examined for their noninvasive capability in brain microvascular endothelial cells (BMEC). The noninvasive mutants exhibited the invasive ability of <1% of the parent strain. One of the noninvasive mutants (10A-23) with a single TnphoA insertion and no changes in phenotypic characteristics was found to be significantly less invasive into the CNS in the newborn rat model of hematogenous E. coli meningitis. The TnphoA inserts with flanking sequences were cloned and sequenced. A novel open reading frame (8.2 kDa) was identified. Open reading frame analysis indicated that the 8.2-kDa protein (Ibe10) contained multiple transmembrane domains. ibe10 was cloned into an expression vector, pQE30, and the purified Ibe10 was shown to inhibit invasion of BMEC by strain RS218. These findings indicate that ibe10 is one of the E. coli genes involved in the invasion of BMEC in vitro and in vivo.

Original languageEnglish (US)
Pages (from-to)4470-4475
Number of pages6
JournalInfection and Immunity
Volume63
Issue number11
StatePublished - 1995
Externally publishedYes

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Molecular Cloning
Escherichia coli Meningitis
Endothelial Cells
Escherichia coli
Brain
Open Reading Frames
Genes
Central Nervous System
Blood-Brain Barrier
Mutagenesis
In Vitro Techniques
Proteins

ASJC Scopus subject areas

  • Immunology

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Escherichia coli invasion of brain microvascular endothelial cells in vitro and in vivo : Molecular cloning and characterization of invasion gene ibe10. / Huang, S. H.; Wass, C.; Fu, Q.; Prasadarao, N. V.; Stins, Monique; Kim, Kwang Sik.

In: Infection and Immunity, Vol. 63, No. 11, 1995, p. 4470-4475.

Research output: Contribution to journalArticle

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AU - Stins, Monique

AU - Kim, Kwang Sik

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