THE PROJECT'S BROAD, LONG TERM OBJECTIVES are to clarify the role in acute bacterial cystitis of four defined virulence factors (VFs) of Escherichia coli both, singly and in combination with one another, and to develop an intervention to prevent cystitis by interfering with the function of one of these VFs.
SPECIFIC AIMS are: 1) to refine a standard mouse model of UTI for the study of cystitis; 2) to use molecular methods for in vitro mutagenesis and allelic exchange to create isogenic VF-deficient mutants of a virulent wild-type E. coli urosepsis isolate; 3) to use these isogenic mutants in the mouse model to determine the contribution to cystitis of specific VFs singly and in combination with one another; 4) to purify from natural sources an inhibitor of one of the four VFs (P fimbriae); and 5) to evaluate the ability of this inhibitor to prevent adherence of E. coli to epithelial cells from women with frequent UTI's and to protect mice from experimental cystitis. EXPERIMENTAL DESIGN AND METHODS. Modified inoculation techniques will be used to avoid nonphysiological vesicoureteral reflux and renal trauma in the mouse model of cystitis. A suicide plasmid system will be used to introduce mutagenized VF determinants into a virulent wild-type E. coli strain in exchange for functional wild-type alleles. The virulence of the resulting isogenic mutants will be compared in the mouse model to reveal the importance of each VF and of combined VFs in the pathogenesis of cystitis. The P1-antigen-containing carbohydrate component of pigeon ovomucoid will be isolated using biochemical methods and tested for its ability 1) to block the adherence of P fimbriated E. coli to human urinary and vaginal epithelial cells from women with frequent UTIs and 2) to protect mice from cystitis due to P fimbriated E. coli. THE HEALTH RELATEDNESS OF THE PROJECT lies in the need for better ways to prevent acute bacterial cystitis, a major health problem of women that is responsible for tremendous morbidity and increased health care costs. Efforts to protect women from bacterial cystitis by interfering with E. coli VFs will be furthered by 1) identifying the specific VFs most responsible for cystitis (which would help guide the future development of anti-VF interventions), and 2) developing an affordable and effective anti-adhesin intervention for the prevention of cystitis.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK047504-03
Application #
2147171
Study Section
Diabetes, Endocrinology and Metabolic Diseases B Subcommittee (DDK)
Project Start
1993-09-30
Project End
1997-08-31
Budget Start
1995-09-15
Budget End
1996-08-31
Support Year
3
Fiscal Year
1995
Total Cost
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
168559177
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Johnson, James R; Clabots, Connie; Rosen, Henry (2006) Effect of inactivation of the global oxidative stress regulator oxyR on the colonization ability of Escherichia coli O1:K1:H7 in a mouse model of ascending urinary tract infection. Infect Immun 74:461-8
Johnson, James R; Jelacic, Srdjan; Schoening, Laura M et al. (2005) The IrgA homologue adhesin Iha is an Escherichia coli virulence factor in murine urinary tract infection. Infect Immun 73:965-71
Johnson, James R; Scheutz, Flemming; Ulleryd, Peter et al. (2005) Host-pathogen relationships among Escherichia coli isolates recovered from men with febrile urinary tract infection. Clin Infect Dis 40:813-22
Johnson, James R; Kuskowski, Michael A; O'bryan, Timothy T et al. (2005) Virulence genotype and phylogenetic origin in relation to antibiotic resistance profile among Escherichia coli urine sample isolates from Israeli women with acute uncomplicated cystitis. Antimicrob Agents Chemother 49:26-31
Manges, Amee R; Johnson, James R; Riley, Lee W (2004) Intestinal population dynamics of UTI-causing Escherichia coli within heterosexual couples. Curr Issues Intest Microbiol 5:49-57
Sannes, Mark R; Kuskowski, Michael A; Johnson, James R (2004) Geographical distribution of antimicrobial resistance among Escherichia coli causing acute uncomplicated pyelonephritis in the United States. FEMS Immunol Med Microbiol 42:213-8
Sannes, Mark R; Kuskowski, Michael A; Johnson, James R (2004) Antimicrobial resistance of Escherichia coli strains isolated from urine of women with cystitis or pyelonephritis and feces of dogs and healthy humans. J Am Vet Med Assoc 225:368-73
Johnson, James R; O'Bryan, Timothy T (2004) Detection of the Escherichia coli group 2 polysaccharide capsule synthesis Gene kpsM by a rapid and specific PCR-based assay. J Clin Microbiol 42:1773-6
Russo, Thomas A; Johnson, James R (2003) Medical and economic impact of extraintestinal infections due to Escherichia coli: focus on an increasingly important endemic problem. Microbes Infect 5:449-56
Johnson, James R; Kaster, Nicholas; Kuskowski, Michael A et al. (2003) Identification of urovirulence traits in Escherichia coli by comparison of urinary and rectal E. coli isolates from dogs with urinary tract infection. J Clin Microbiol 41:337-45

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