MICROBAL UREASE--A POTENTIAL ANTIGEN MIMIC OF HLA-B27

这些项目的目标是评估分子模拟作为一种 HLA B27相关疾病的发病机制，并检查 这些疾病的潜在动物模型。 这些疾病中有几种可以 导致人类关节病， 条件 目前的研究表明，某些微生物尿素酶可能 作为HLA-B27的分子模拟物，因为尿素酶的α亚基 来自解脲脲原体，肺炎克雷伯氏菌，幽门螺杆菌， 小肠结肠炎耶尔森菌和假结核耶尔森菌与抗- B27单克隆抗体（MAb）。 U.尿素分解产生 最强烈的反应。 天然尿素酶（Uu 150 K）来源于U.解脲支原体将是 纯化，并且将尿素酶α亚基（pep 243）的区域 合成并纯化。 这两种分子都将作为一种 HLA-B27转基因小鼠（TGM）的分子模拟物。 的某些菌株 这些小鼠有可能成为研究关节炎的动物模型。 炎症和关节炎特征性B27相关疾病。 到 为了实现本项目的目标，具体目标是：1）确定 B27-TGM的选择菌株的组织中的B27表达， 组织切片免疫细胞化学分析; 2）鉴定交叉 抗pep 243单克隆抗体和HLA-B27之间的反应性表达的细胞， 筛选B27_ TGM菌株; 3）检测细胞，纯化Uu 150 k，pep 243， 或H2 B6免疫原作为测试抗原; 4）检查B27 TGM和对照， 新合成抗体与特定组织的结合 用测试抗原接种; 5）评估抗- pep 243 MAb对B27+TGM的特异性组织的作用 这些抗原，以确定是否新的和对照小鼠血清的抗- B27抗Uu 150 k和抗pep 243抗体，接种后 测试抗原。 将获得的数据用于评价微生物 尿素酶作为B27相关疾病的发病机制， 为潜在动物提供某些基线免疫细胞化学数据 研究这些疾病的模型，并测试潜在的 动物模型

The goals of the projects are to evaluate molecular mimicry as a mechanism of pathogenesis for HLA B27-associated diseases, and to examine potential animal models for these disease. Several of these diseases can result in arthropathies in humans, and lead to chronic disabling conditions. Present studies suggest that certain microbial ureases may act as a molecular mimic of HLA-B27, since the alpha subunit of ureases from Ureaplasma urealyticum, Klebsiella pneumoniae, Helicobacterpylori, Yersinia enterocolitica and Y, pseudotuberculosis cross-react with anti- B27 monoclonal antibody (MAb). Urease from U. urealythicum yields the strongest reaction. Native urease (Uu150K) from U. urealyticum will be purified, and a region of the urease alpha subunit (pep243) will be synthesized and purified. Both of these molecules will be tested as a molecular mimic of HLA-B27-transgenic mice (TGM). Certain strains of these mice have the potential to be animal model for studies of joint inflammation and arthritis characteristic B27-associated disease. To achieve the goals of this project, the specific aims are to 1) identify B27 expression in tissues from select strains of B27-TGM, using immunocytochemical analysis of histological sections; 2) identify cross reactivity between anti-pep243 MAb and HLA-B27 expressed on the cells of select strains of B27_ TGM; 3) examine cells, purified Uu150k, pep243, or H2B6 immunogen as test antigens; 4) examine B27 TGM and controls for the binding of newly synthesize antibody to specific tissues following inoculation with the test antigens; 5) assess binding inhibition of anti- pep243 MAb to specific tissues of B27+TGM following inoculation with the these antigens, to determine if the newly and control mice sera for anti- B27 anti-Uu150k, and anti-pep 243 antibodies, following inoculation with the test antigens. The data to be obtained will evaluate microbial urease as a mechanism of pathogenesis for B27 associated diseases, provide certain baseline immunocytochemical data for potential animal models to study these diseases, and test the efficacy of the potential animal models.