ROLE OF BACTERIAL LECTINS IN KIDNEY DISEASES

细菌凝集素在肾脏疾病中的作用

• 批准号: 2838112
• 负责人: Bogdan J Nowicki
• 金额: $ 15.84万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-02-01 至 2000-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2838112
• 关键词: CHO cells; Escherichia coli; SCID mouse; adhesin; antigen receptors; bacterial antigens; bacterial cytopathogenic effect; chromatography; disease /disorder model; enzyme linked immunosorbent assay; epitope mapping; host organism interaction; interstitial cystitis; lectin; microorganism hemagglutinin; molecular cloning; mutant; pilus; pyelonephritis; receptor binding; urinary tract infection

DESCRIPTION (Adapted from the applicant's abstract): The hypothesis proposed in grant R01 DK42029 stated that renal interstitial attachment of Dr adhesin bearing Escherichia coli creates structural basis for development of chronic interstitial nephritis. Exploration of consequences of Dr-adhesin Dr-tissue-ligand interaction resulted in the development of an experimental model of ascending chronic pyelonephritis (ChP) that fulfills histological requirements for human ChP. In this project, the investigators will attempt to understand the molecular mechanisms involved in and to prevent the development of ChP. They will attempt the following projects. Map receptor-binding epitopes on short consensus repeat-3 (SCR-3) domain of decay accelerating factor (DAF) for Dr fimbriae. (a) Map Dr binding epitopes on DAF-SCR-3 by synthetic peptides by competitive inhibition. (b) Map Dr binding epitopes on DAF-SCR-3 by antipeptide antibodies. (c) Replace single amino acids within the SCR-3 region using site-directed mutagenesis. (d) Test in vitro attachment of BN406 on Chinese hamster ovary (CHO) cells that express constructed DAF (SCR-3) mutants. (2) Map functional regions within the dra operon necessary for interstitial colonization and developing ChP on a mouse C3H/HeJ in vivo model. (a) Characterize the nucleotide sequence of the draA, draB. draC, draD, and draE mutants on a ChP model to dissect involvement of adhesive, and renal interstitial tropism properties in the development of ChP. (c) Examine virulence of draA substitution mutants defective in type-IV collagen binding on a ChP mouse model. (d) Study route and kinetics of dra mutants spread leading to interstitial colonization on a SCID mouse model. (3) Test the effect of vaccination with recombinant Dr adhesin on development of ChP in C3H/HeJ mice.

描述（改编自申请人的摘要）：假设 拨款 R01 DK42029 中提出的建议指出，肾间质附着 大肠杆菌博士粘附素为发育创造结构基础 慢性间质性肾炎。 探索后果 Dr-adhesin Dr-组织-配体相互作用导致了 上行性慢性肾盂肾炎（ChP）实验模型，满足 人类 ChP 的组织学要求。 在这个项目中，研究人员 将尝试了解所涉及的分子机制并 预防 ChP 的发展。 他们将尝试以下项目。 将受体结合表位绘制在短共有重复序列 3 (SCR-3) 结构域上 Dr fimbriae 的衰变加速因子 (DAF)。 (a) Map Dr绑定 通过竞争性抑制合成肽来抑制 DAF-SCR-3 上的表位。 (二) 通过抗肽抗体绘制 DAF-SCR-3 上 Dr 结合表位的图谱。 (c) 替换 使用定点诱变对 SCR-3 区域内的单个氨基酸进行突变。 (d) 体外测试 BN406 对中国仓鼠卵巢 (CHO) 细胞的附着 表达构建的 DAF (SCR-3) 突变体。 (2) 地图功能区 在间质定殖和发育所必需的 dra 操纵子内 小鼠 C3H/HeJ 体内模型上的 ChP。 (a) 表征核苷酸 draA、draB的序列。 ChP 模型上的 draC、draD 和 draE 突变体 解剖涉及粘附性和肾间质向性特性 在 ChP 的发展中。 (c) 检查 draA 取代的毒力 ChP 小鼠模型上 IV 型胶原蛋白结合缺陷的突变体。 (四) 研究dra突变体传播导致间质的途径和动力学 SCID 小鼠模型上的定植。 (3) 检验疫苗接种效果 重组 Dr 粘附素对 C3H/HeJ 小鼠 ChP 发育的影响。