Studies of Urinary Tract infection reveal an increase in incidence in the 20-40 year old female associated with sexual activity and pregnancy. Even more striking is the progressive increase in incidence after menopause, to the extent that one-third of the female population over 70 years of age have bacteria.
The aim of this proposal is elucidate the mechanism of bacterial adherence and colonization in the urinary bladder. This will include defining the bladder's normal anti-adherence properties and the events leading t their failure in some circumstances (e.g., aging, hormonal deprivation). It has been shown that a glycoprotein lining the bladder probably acts as a bacterial anti-adherence substance since its removal leads to increased bacterial adherence and colonization. A glycoprotein fraction (GPI) isolated from rabbit bladders was used to develop antisera in mice. These sera bind to the mucin layer of rabbit, guinea pig, rat, hamster and human bladders and can be used in quantitative assays (ELISA and RIA) to detect the glycoprotein in urine. As there is only indirect evidence that the glycoprotein layers acts as an antibacterial substance in the bladder, the first objective of this study is to determine whether this glycoprotein can prevent bacterial attachment to uroepithelial cells. This will be done by attempting to reconstitute an antibacterial state in mucin deficient bladders (acid treated or oophorectomized) by infusion of isolated glycoprotein. Subsequently, an in vitro, quantitated assay measuring bacterial adherence to single cell suspensions of uroepithelial cells will be developed. The ability of the GPI fraction to inhibit this binding will be measured and its interaction with isolated epithelial cells and bacteria quantitated. At the same time the complete biochemical characterization of GPI will be done. This will include physicochemical characterization (e.g., molecular weight, subunit structure, carbohydrate and amino analysis composition) as well as the identification of those moieties involved in anti-adherence and anti-genicity. This will be done with fragments obtained from enzymatic and chemical degradation. The second phase of this study is to determine the distribution of the glycoprotein in the human urinary tract by immunohistochemical procedures and to quantitate its level in the urine (ELISA, RIA). We will measure the urinary levels of the human glycoprotein in healthy volunteers and patients with urological disorders (chronic urinary tract infection, interstitial cystitis).
| Byrne, D E; Mac Phee, M; Mulholland, M et al. (1994) Urinary tract glycoprotein: distribution and antigenic specificity. World J Urol 12:21-6 |
| Callahan, H J; Shupp-Byrne, D; Pizzo, M et al. (1991) The production of antibodies to pentosanpolysulfate (ELMIRON, SP-54). J Immunol Methods 136:53-9 |
| Stefanelli, J; Callahan, H J; Byrne, D S et al. (1990) Antisera to a rabbit urinary tract antigen also react with human bladder and kidney tissue. J Urol 143:414-7 |
