ROLE OF BACTERIAL LECTINS IN KIDNEY DISEASES

描述(改编自申请人的摘要)：假设 在授予R01 DK42029的建议中指出，肾间质附着 携带粘附素的大肠埃希菌为发育创造结构基础 慢性间质性肾炎。对以下问题后果的探讨 DR-粘附素DR-组织-配体相互作用导致 实验性慢性肾盂肾炎(CHP)模型的建立 人慢性胰腺炎的组织学要求。在这个项目中，调查人员 将试图理解涉及的分子机制和 防止慢性精神分裂症的发展。他们将尝试以下项目。 将受体结合表位映射在猪瘟病毒SCR-3结构域上 菌毛衰变加速因子(DAF)。(A)MAP DR装订 竞争抑制合成肽在DAF-SCR-3上的表位。(B) 用抗肽抗体定位DAF-SCR-3上的DR结合表位。(C)更换 在SCR-3区域使用定点突变的单一氨基酸。 (D)BN406对中国仓鼠卵巢(CHO)细胞的体外贴壁实验 表达构建的DAF(Scr-3)突变体。(2)地图功能区 在间质定植和发育所需的DRA操纵子内 CHP对C3H/HeJ小鼠体内模型的影响。(A)确定核苷酸的特征 德拉拉的序列，单调乏味。DRAC、DRAD和DRE突变体在CHP模型上 解剖粘连受累与肾间质的趋向性 在CHP的发展过程中。(C)检查DRAA替代的毒性 CHP小鼠模型的IV型胶原结合缺陷突变体。(D) Dra突变体向间质扩散的研究路线和动力学 在SCID小鼠模型上的定植。(3)测试疫苗接种效果 重组DR黏附素对C3H/HeJ小鼠慢性胰腺炎发生的影响

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.