Development of Vaccines Against Uropathogenic Escherichia coli Toxins to Reduce Morbidity Associated with Urinary Tract Infections
Abstract
Urinary tract infections (UTIs) are estimated to affect eighty percent of women in the United States annually with a resulting cost of billions of dollars spent on physician visits, treatment, and lost work hours. Uropathogenic E. coli (UPEC) are the leading cause of UTIs. Previous research in our laboratory demonstrated that female C3H/HeOuJ mice that are transurethrally infected with UPEC strain CP9 into the urinary bladder develop a suppurative cystitis with severe submucosal edema within 24 hours of bacterial inoculation. Researchers in our laboratory also reported that CP9 strains with inactivated Cytotoxic Necrotizing Factor 1 (CNF1) and/or the Hemolysin (HlyA) genes evoke less severe pathology in the urinary bladder than does the wild type strain. CNF1 and HlyA are toxins that contribute to UPEC pathogenesis. CNF1 enzymatically causes constitutive activation of small Rho family GTPases that leads to cytoskeletal changes in host cells. HlyA forms membrane pores and lyses mammalian cells. CNF1 and HlyA contribute to the robust inflammatory response in the bladders of mice challenged for 24hrs with the UPEC strain CP9. We hypothesized that antibodies against CNF1 and/or HlyA will reduce the pathology caused by CP9 infection of the bladder. Therefore, we actively immunized mice with a HlyA toxoid in which two essential lysine residues were changed to glutamic acid. Female C3H/HeOuJ mice were vaccinated subcutaneously twice with HlyA toxoid in adjuvant and boosted once or twice more without adjuvant. We collected serum prior to each boost, and we observed increases in anti-HlyA titers over time by ELISA and elevations in neutralizing antibody levels as assessed by erythrocyte lysis neutralization after the second boost. Two weeks after the final boost, we challenged the mice intraurethrally with CP9 and sacrificed them 24 hr later. We observed a reduction in urine
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 03, 2014
- Accession Number
- AD1127840
Entities
People
- Mark A. Smith
Organizations
- Uniformed Services University of the Health Sciences