BLADDER GLYCOSPHINGOLIPIDS IN THE PATHOGENESIS OF URINARY TRACT INFECTION

尽管到目前为止，膀胱是尿路感染最常见的部位，但很少是 已知感染细菌附着在其中的分子 组织与肾脏相比，其中球状细胞在GSLS中的作用 作为细菌对大肠埃希菌的附着部位已经有了很好的研究。 由于GSLS的碳水化合物部分在不同的组织中有所不同，通常 在宿主基因控制下，这些分子作为两者的决定因素 患者对感染的易感性和致病的组织嗜性 细菌。该方案总体目标是利用原电池 从膀胱培养作为典范系统来定义作用和 膀胱GSLS作为泌尿系致病菌附着部位的调控， 重点关注导致女性膀胱炎的两个最常见的原因，即大肠杆菌和 在以下拟议的研究中： (1)评估球状序列(GB)和神经节细胞序列的假说 (GG)膀胱移行细胞原代培养中存在GSLs 分别与大肠杆菌和腐生葡萄球菌结合，我们将 从细胞中提纯GSLs，鉴定与大肠杆菌和腐生葡萄球菌的结合 GSLS，并用特定的单抗确认GSLS的身份 抗体(单抗)和形式碳水化合物结构分析； 评估在特定目标1中确定的GSLS是 在原代培养的膀胱细胞中暴露的表面，并具有功能 与大肠杆菌和腐生葡萄球菌的附着相关，我们将研究 细胞培养前和处理后的细菌黏附 对于腐生菌附着，我们将在之前研究细菌黏附 在用GSL抑制剂对细胞培养进行预处理后 针对相关GSLS的合成或单抗；(3)评估 分泌型和ABO基因影响gb表达的假说 和GB GSLS在膀胱中，因此结合大肠杆菌的能力，我们 将建立已知患者的原代膀胱培养 分泌者/ABO状态并确定预测的 培养细胞中存在糖基转移酶和GSL产物； 以及(4)评估细胞因子从原代细胞释放的假说 培养的尿路上皮细胞，并调节E.Coli-和S. 腐植酸结合的GSLS在膀胱中，我们将检查其影响 大肠埃希菌和腐生链霉菌对细胞因子释放的影响 细胞及肿瘤坏死因子和IL-1、IL-6、IL-8的作用 原代培养的尿路上皮细胞相关GSLs的表达及意义 细菌附着在这些细胞上。这些关于基因表达和表达的研究 膀胱GSLS的调节与感染的发病机制有关， 该器官的炎症和分化，并最终可能导致 涉及防止细菌附着和感染的新方法。

Although the bladder is by far the most common site of UTI, little is known about the molecules to which infecting bacteria attach in this tissue as compared with the kidney, in which the role of globoseries GSLs as bacterial attachment sites for Escherichia coli has been well studied. Since the carbohydrate portion of GSLs varies in different tissues, often under host genetic control, these molecules serve as determinants of both patient susceptibility to infection and tissue tropism for pathogenic bacteria. The overall goal of this proposal is to utilize primary cell cultures from the bladder as a model system to define the role and regulation of bladder GSLs as attachment sites for uropathogenic bacteria, focusing on the two most common causes of cystitis in women, E. coli and S. saprophyticus, in the following proposed studies: (1) To evaluate the hypothesis that globoseries (Gb) and ganglioseries (Gg) GSLs are present in primary cultures of bladder transitional epithelium and bind E. coli and S. saprophyticus, respectively, we will purify GSLs from the cells, identify E. coli-and S. saprophyticus-binding GSLs, and confirm the identities of the GSLs using specific monoclonal antibodies (MAbs) and formal carbohydrate structural analysis; (2) To evaluate the hypothesis that the GSLs identified in Specific Aim 1 are surface exposed in primary bladder cell cultures and are functionally relevant in E. coli and S. saprophyticus attachment, we will study bacterial adherence before and after pre-treatment of the cell cultures with an saprophyticus attachment, we will study bacterial adherence before and after pre-treatment of the cell cultures with an inhibitor of GSL synthesis or MAbs directed against relevant GSLs; (3) To evaluate the hypothesis that the secretor and ABO genes influence the expression of Gb and Gb GSLs in the bladder and thus the capacity for E. coli binding, we will establish primary bladder cultures from patients of known secretor/ABO status and determine whether the predicted glycosyltransferases and GSL products are present in the cultured cells; and (4) To evaluate the hypothesis that cytokines are release from primary uroepithelial cell cultures and modulate the expression of E.coli-and S. saprophyticus-binding GSLs in the bladder, we will to examine the effects of adherence by E. coli and S. saprophyticus on cytokine release from the cells and the effects of tumor necrosis factor and IL-1, IL-6 and IL-8 on the expression of relevant GSLs in primary urothelial cell cultures and on bacterial adherence to these cells. These studies on the expression and regulation of bladder GSLs are relevant to the pathogenesis of infection, inflammation and differentiation in this organ and could eventually lead to novel approaches to prevention of bacterial attachment and infection.