Number : DAMD 1796-1-6301 TITLE : Novel Approaches to Preventing Urinary Tract Infection in Women

1. Gupta K, Hooton TM, Stapleton AE, Roberts P, Winter C, Deshaw N, Wobbe C, Fennell C, Denton A, Kahn JB, Stamm WE. Efficacy of self-diagnosis and self-treatment for management of uncomplicated This report contains unpublished data that should be protected Final Report for DAMD17-96-1-6301 Stapleton, Ann E. MD 14 recurrent urinary tract infections in women. Program and abstracts of the 36th annual meeting of the Infectious Diseases Society of America, abstract no. 39. 2. Lemley CA, Ballweber LM, Johnson ML, Kiselev AO, Stamm WE, Stapleton A, Lampe MF. Identification of putative Chlamydia binding proteins from HeLa cells and immortalized human cervical cells. The American Federation for Medical Research, Western section annual meetings. Carmel, California, February 1999. Abstracts. 3. Grady RW, Mitchell ME, Stapleton AE. High-throughput analysis of differential gene expression of in vitro urothelium exposed to uropathogenic Escherichia coli pDC-1. Program and abstracts of the 1999 annual meeting, American Urological Association, abstract no. 21. 4. Grady RW, Mitchell ME, Mahmoudi A, Stapleton AE. Expression array "gene chip" analysis of in vitro urothelium exposed to uropathogenic Escherichia coli pDC-1. Presented at the annual meeting of the American Academy of Pediatrics, 1999. 5. Patton DL, Agnew KJ, Meier A, Aura J, Hooton T, Stapleton A, Stamm WE. Influence of a single episode of intercourse on the vaginal flora and epithelium with and without condom use. Presented at the Infectious Diseases Society of Obstetrics and Gynecology, 1999. 6. Schlager TA, Whittam TS, Hendley JO, Wilson RA, Bhang J, Grady R, Stapleton A. Expression of virulence factors among Escherichia coli isolated from periurethra and urine of children with neurogenic bladder on intermittent catheterization. Proceedings of the Cyokines/Chemokines In Infectious Diseases Meeting, National Institutes of Health, September 8-10, 1999, Bethesda, Maryland (Poster #055). 7. Schlager TA, Whittam TS, Hendley JO, Wilson RA, Bhang J, Grady R, Stapleton A. Carriage of Escherichia coli and expression of virulence factors in urine and periurethra of patients with neurogenic bladder on intermittent catheterization. Proceedings of the International Bladder Symposium, November 4-7, 1999, Washington DC. 8. Yarova-Yarovaya Y, Waitzman S, Atala A, Yoo J, Stroud M, Grady R, Stapleton A. Bladder glycosphingolipids in urinary tract infection. Proceedings of the International Bladder Symposium, March 8-11, 2001, Washington DC. Presentations and Meetinns "* Moderator and Scientific Committee, 2000 and 2001 International Bladder Symposia "* Invited Speaker, Second International Bladder Research Congress, San Francisco, April 1998. "• Invited Speaker, Rodin Conference, July 1997 and July 2000 "• Invited speaker, The Millennium Symposium of Pyelonephritis and UTI, Lund, Sweden, May 2000 "* Invited speaker, University of Washington Science in Medicine New Investigator series, 1999-2000 (also received award in connection with this lecture series) This report contains unpublished data that should be protected Final Report for DAMD 17-96-1-6301 Stapleton, Ann E. MD 15 • Obstetrics and Gynecology Grand Rounds, 2000 Poster presentation, 2001 International Bladder Symposium Development of cell lines Primary cultures of vaginal epithelial cells derived from individuals have been developed for the purposes of these studies. However, these cells have not been provided to other investigators as a "reagent" and are not suitable for an application of that nature, such as submission to a cell banking facility. Personnel supported by this grant Ann Stapleton, MD Mark Stroud, PhD Amy Denton, BS Michelle Brown, BS Afshin Mahmoudi, MS 9. CONCLUSIONS and "so what" comments Our initial years of funding were focused on refining the techniques we developed for culturing primary vaginal epithelial cells and on adapting Dr. Atala's methods for culturing primary human urothelial cells to our model system. We have now established these in vitro models of primary cultured bladder epithelial and human vaginal epithelial cells for the identification, purification and structural characterization of E. coliand S. saprophvticus-binding glycosphingolipid moieties. These models will be valuable for future research in UTI and other urogenital infections and will also contribute to basic knowledge about the bladder and the vagina. We have completed studies characterizing bacterial adherence to primary bladder and vaginal epithelial cells utilizing organisms that are key uropathogens for healthy young women, E. coli and S. saprophyticus. Findings in these systems will also be applicable to the study of other mucosal infections with E. coli and other gram negative pathogens, and may also be applicable to studies of Staphylococcus aureus. We have also established that GSLs are essential for the adherence of pa. -adhesin-expressing uropathogenic E. coli in these systems. We have completed characterization of primary cultured vaginal epithelial cells according to parameters normally applied to similar epithelia, in collaboration with respected investigators in the field of epithelial biology. We have also characterized the effects of exogenous estrogen on the expression of GSLs and keratins as well as on bacterial adherence in vaginal epithelial cells, all of which are relevant to the use of these cells as models of UTI and other urogenital diseases throughout the life cycle. This model of the vaginal epithelium provides new opportunities for studying other aspects of women's urogenital health, including testing vaginal products, probiotics, and contraceptives; understanding protective roles of organisms in the normal flora; and studying the cellular effects of hormone replacement therapy on a key target tissue, the vagina. Lastly, we have advanced the study of the globoseries GSLs, which have not been thoroughly studied in normal adult tissues. Thus, our findings have additional relevance in the field of glycobiology.