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 人类白细胞抗原B27相关疾病发病机制的研究 这些疾病的潜在动物模型。其中几种疾病可能 导致人类关节疾病，并导致慢性残疾 条件。目前的研究表明，某些微生物尿素酶可能 作为人类白细胞抗原-B27的分子模拟物，因为尿素酶的阿尔法亚单位 来自解脲支原体、肺炎克雷伯菌、幽门螺杆菌、 小肠结肠炎耶尔森菌和假结核耶尔森菌与抗-HBs的交叉反应 B27单抗(MAb)。来自解脲支原体的尿素酶产生 最强烈的反应。解脲支原体的天然尿素酶(Uu150K)将是 纯化后，尿素酶α亚基的一个区域(Pep243)将被 合成和提纯。这两个分子都将作为一个 人类白细胞抗原B27转基因小鼠(TGM)的分子模拟某些菌株 这些小鼠有可能成为关节研究的动物模型。 炎症和关节炎是B27相关疾病的特征。至 实现本项目的目标，具体目标是：1)确定 B27-TGM筛选株组织中B27的表达 组织切片的免疫细胞化学分析；2)鉴定交叉 抗肽243单抗与人类白细胞抗原B27表达的反应性 选择B27_TGM菌株；3)细胞检测，纯化的Uu150k，Pep243， 或H_2B6免疫原作为试验抗原；4)检测B27 TGM和对照 新合成的抗体与特定组织的结合 用测试抗原接种；5)评估抗-DNA结合抑制 免疫B27 TGM后对特定组织的Pep243单抗 这些抗原，用来确定新生小鼠和对照小鼠的血清是否具有抗 B27抗Uu150k和抗PEP 243抗体 测试抗原。将获得的数据将评估微生物 尿素酶作为B27相关疾病的发病机制， 为潜在动物提供特定的基线免疫细胞化学数据 研究这些疾病的模型，并测试潜在的疗效 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.