ROLE OF BACTERIAL LECTINS IN KIDNEY DISEASES

描述（改编自申请人摘要）：假设 R 01 DK 42029中提出的研究表明， 携带粘附素的大肠杆菌博士为开发创造了结构基础 慢性间质性肾炎 探索的后果 DR-粘附素DR-组织-配体相互作用导致了 上行性慢性肾盂肾炎（ChP）的实验模型， 人ChP的组织学要求。 在这个项目中，研究人员 将试图了解参与的分子机制， 防止ChP的发展。 他们将尝试以下项目。 在短共有重复序列-3（SCR-3）结构域上定位受体结合表位 菌毛博士的衰变加速因子（decay accelerating factor，ERF）。 (a)Map Dr绑定 通过竞争性抑制合成肽对DAF-SCR-3上的表位进行修饰。 （B） 通过抗肽抗体定位DAF-SCR-3上的Dr结合表位。 (c)取代 使用定点诱变在SCR-3区域内的单个氨基酸。 (d)BN 406在中国仓鼠卵巢（CHO）细胞上的体外附着试验 其表达构建的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， ChP对小鼠C3 H/HeJ体内模型的作用。 (a)鉴定核苷酸 draA、draB的顺序。 在ChP模型上的draC、draD和draE突变体， 粘连和肾间质向性的解剖参与 在ChP的发展中。 (c)检查draA置换的毒力 在ChP小鼠模型上IV型胶原结合缺陷的突变体。 （d）其他事项 研究路线和动力学的dra突变传播导致间质 在SCID小鼠模型上的定殖。 (3)测试接种疫苗的效果， 重组Dr粘附素对C3 H/HeJ小鼠体内ChP发育的影响。

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.