Proteus mirabilis commonly infects the complicated urinary tract and is associated with urolithiasis. Stone formation is caused by bacterial urease, which hydrolyzes urea to ammonia, causing local pH to rise, and leads to the subsequent precipitation of magnesium ammonium phosphate (struvite) and calcium phosphate (apatite) crystals. To prevent these infections, we vaccinated CBA mice with formalin-killed bacteria or purified mannose-resistant, Proteus-like (MR/P) fimbriae, a surface antigen expressed by P. mirabilis during experimental urinary tract infection, via four routes of immunization: subcutaneous, intranasal, transurethral, and oral. We assessed the efficacy of vaccination using the CBA mouse model of ascending urinary tract infection. Subcutaneous or intranasal immunization with formalin-killed bacteria and intranasal or transurethral immunization with purified MR/P fimbriae significantly protected CBA mice from ascending urinary tract infection by P. mirabilis (P < 0.05). To investigate the potential of MrpH, the MR/P fimbrial tip adhesin, as a vaccine, the mature MrpH peptide (residues 23 to 275, excluding the signal peptide), and the N-terminal receptor-binding domain of MrpH (residues 23 to 157) were overexpressed as C-terminal fusions to maltosebinding protein (MBP) and purified on amylose resins. Intranasal immunization of CBA mice with MBPMrpH (residues 23 to 157) conferred effective protection against urinary tract infection by P. mirabilis (P < 0.002).The urinary tract is a complicated epithelium-lined tube with an opening to the body surface. It is susceptible to infections by exogenous organisms that can colonize the peri-urethral area, enter the bladder via the urethra, and ascend the ureters to the kidneys. In some cases the organisms can enter the bloodstream and cause sepsis. Most urinary tract infections (UTIs) occur in otherwise healthy women with normal urinary tracts, but a significant proportion of UTIs develop in those with complicated urinary tracts, including those that are catheterized or otherwise instrumented or obstructed due to structural abnormalities that prevent the normal flow of urine (42).Proteus mirabilis infects a high proportion of patients with complicated urinary tracts (29, 44). Importantly, once in the urinary tract, the bacterium appears to have a predilection for the kidney (10). Furthermore, this bacterium causes not only cystitis and acute pyelonephritis (9, 11, 13, 38, 41) but also urinary stones (14), further complicating already abnormal urinary tracts.Stone formation is caused by the expression of bacterial urease, which hydrolyzes urea to ammonia, causing local pH to rise, and leads to the subsequent precipitation of magnesium ammonium phosphate (struvite) and calcium phosphate (apatite) crystals (14,31,32). The stones resulting from aggregation of such crystals begin forming at the surface of the epithelium (25), which complicates infection for three reasons. First, the P. mirabilis organisms caught within the interstices of the forming stones are ver...