The principal investigator has established an infant rat model of experimental hematogenous meningitis, which mimics human E. coli meningitis (i.e., hematogenous infection of the meninges without the need for adjuvant or direct inoculation of bacteria into cerebrospinal fluid). They have established an in vitro model of blood-brain barrier using brain microvascular endothelial cells (BMEC). Using both in vitro and in vivo systems and the BMEC model, they have shown that successful traversal of E. coli across the blood-brain barrier may be a complex process involving separate steps of E. coli-BMEC interactions, i.e., binding to BMEC and invasion of BMEC. They have shown that S fimbriae are the major E. coli structures contributing to binding to BMEC. They therefore hypothesize that the role of S fimbriae is to have a more intimate contact for circulating E. coli to BMEC to withstand blood flow, which may be required for subsequent crossing of the blood-brain barrier, but there is no information to support this hypothesis. The overall aim of the proposal is to study the role of binding via S fimbriae in the pathogenesis of E. coli meningitis, both in vitro (using BMEC in culture) and in vivo (using an experimental animal model of hematogenous E. coli meningitis that closely mimics the pathogenesis of human E. coli meningitis).
The specific aims are: (1)to continue to construct isogenic S fimbriae operon deletion (delta sfaII) mutant of invasive E. coli K1 strain RS218 by chromosomal gene replacement (allelic exchange); (2)to examine the ability of S fimbriae negative (delta sfaII) mutant to bind and invade brain microvascular endothelial cells (BMEC) in vitro and in vivo; (3)to determine whether it is possible to restore the ability of S fimbriae negative (delta sfaII) mutant to bind and invade BMEC by complementation with the cloned sfaII gene cluster; (4)to assess which specific subunits of S fimbriae are responsible for binding to BMEC by constructing single gene deletion mutants (delta sfaIIS, delta sfaIIA, delta sfaIIG, delta sfaIIH); (5)and to identify and characterize BMEC glycoprotein(s) interactive with S fimbriae.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI047225-04
Application #
6511245
Study Section
Special Emphasis Panel (ZRG1-BM-1 (01))
Program Officer
Korpela, Jukka K
Project Start
2000-04-01
Project End
2004-03-31
Budget Start
2002-04-01
Budget End
2003-03-31
Support Year
4
Fiscal Year
2002
Total Cost
$327,000
Indirect Cost
Name
Johns Hopkins University
Department
Pediatrics
Type
Schools of Medicine
DUNS #
045911138
City
Baltimore
State
MD
Country
United States
Zip Code
21218
Kim, Kwang S (2012) Current concepts on the pathogenesis of Escherichia coli meningitis: implications for therapy and prevention. Curr Opin Infect Dis 25:273-8
Shin, Sooan; Kim, Kwang Sik (2010) Human brain endothelial ATP synthase beta-subunit is mannose-insensitive binding target of FimH. FEMS Microbiol Lett 303:156-62
Yao, Yufeng; Xie, Yi; Perace, Donna et al. (2009) The type III secretion system is involved in the invasion and intracellular survival of Escherichia coli K1 in human brain microvascular endothelial cells. FEMS Microbiol Lett 300:18-24
Siddharthan, Venkatraman; Kim, Yuri V; Liu, Suyi et al. (2007) Human astrocytes/astrocyte-conditioned medium and shear stress enhance the barrier properties of human brain microvascular endothelial cells. Brain Res 1147:39-50
Yao, Yufeng; Xie, Yi; Kim, Kwang Sik (2006) Genomic comparison of Escherichia coli K1 strains isolated from the cerebrospinal fluid of patients with meningitis. Infect Immun 74:2196-206
Paul-Satyaseela, Maneesh; Xie, Yi; Di Cello, Francescopaolo et al. (2006) Responses of brain and non-brain endothelial cells to meningitis-causing Escherichia coli K1. Biochem Biophys Res Commun 342:81-5
Xie, Yi; Kolisnychenko, Vitaliy; Paul-Satyaseela, Maneesh et al. (2006) Identification and characterization of Escherichia coli RS218-derived islands in the pathogenesis of E. coli meningitis. J Infect Dis 194:358-64
Jain, Sanjay K; Paul-Satyaseela, Maneesh; Lamichhane, Gyanu et al. (2006) Mycobacterium tuberculosis invasion and traversal across an in vitro human blood-brain barrier as a pathogenic mechanism for central nervous system tuberculosis. J Infect Dis 193:1287-95
Xie, Yi; Yao, Yufeng; Kolisnychenko, Vitaliy et al. (2006) HbiF regulates type 1 fimbriation independently of FimB and FimE. Infect Immun 74:4039-47
Shin, Sooan; Kim, Kwang Sik (2006) RhoA and Rac1 contribute to type III group B streptococcal invasion of human brain microvascular endothelial cells. Biochem Biophys Res Commun 345:538-42

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