Escherichia coli K1 aslA contributes to invasion of brain microvascular endothelial cells in vitro and in vivo

J. A. Hoffman, J. L. Badger, Y. Zhang, S. H. Huang, Kwang Sik Kim

Research output: Contribution to journalArticle

Abstract

Neonatal Escherichia coli meningitis remains a devastating disease, with unacceptably high morbidity and mortality despite advances in supportive care measures and bactericidal antibiotics. To further our ability to improve the outcome of affected neonates, a better understanding of the pathogenesis of the disease is necessary. To identify potential bacterial genes which contribute to E. coli invasion of the blood-brain barrier, a cerebrospinal fluid isolate of E. coli K1 was mutagenized with TnphoA. TnphoA mutant 27A-6 was found to have a significantly decreased ability to invade brain microvascular endothelial cells compared to the wild type. In vivo, 32% of the animals infected with mutant 27A-6 developed meningitis, compared to 82% of those infected with the parent strain, despite similar levels of bacteremia. The DNA flanking the TnphoA insertion in 27A-6 was cloned and sequenced and determined to be homologous to E. coli K-12 aslA (arylsulfatase-like gene). The deduced amino acid sequence of the E. coli K1 aslA gene product shows homology to a well-characterized arylsulfatase family of enzymes found in eukaryotes, as well as prokaryotes. Two additional aslA mutants were constructed by targeted gene disruption and internal gene deletion. Both of these mutants demonstrated decreased invasion phenotypes, similar to that of TnphoA mutant 27A-6. Complementation of the decreased-invasion phenotypes of these mutants was achieved when aslA was supplied in trans. This is the first demonstration that this locus contributes to invasion of the blood-brain barrier by E. coli K1.

Original languageEnglish (US)
Pages (from-to)5062-5067
Number of pages6
JournalInfection and Immunity
Volume68
Issue number9
DOIs
StatePublished - 2000
Externally publishedYes

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Arylsulfatases
Endothelial Cells
Escherichia coli
Brain
Genes
Blood-Brain Barrier
Escherichia coli Meningitis
Phenotype
Bacterial Genes
Gene Deletion
Bacteremia
Eukaryota
Meningitis
Cerebrospinal Fluid
In Vitro Techniques
Amino Acid Sequence
Anti-Bacterial Agents
Morbidity
Mortality
DNA

ASJC Scopus subject areas

  • Immunology

Cite this

Escherichia coli K1 aslA contributes to invasion of brain microvascular endothelial cells in vitro and in vivo. / Hoffman, J. A.; Badger, J. L.; Zhang, Y.; Huang, S. H.; Kim, Kwang Sik.

In: Infection and Immunity, Vol. 68, No. 9, 2000, p. 5062-5067.

Research output: Contribution to journalArticle

Hoffman, J. A. ; Badger, J. L. ; Zhang, Y. ; Huang, S. H. ; Kim, Kwang Sik. / Escherichia coli K1 aslA contributes to invasion of brain microvascular endothelial cells in vitro and in vivo. In: Infection and Immunity. 2000 ; Vol. 68, No. 9. pp. 5062-5067.
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