CHARACTERIZATION OF RHEUMATOID ANTIGENS IN CYANOBACTERIA

描述：(摘自申请者的摘要)几种风湿病 疾病与自身免疫抗体和/或 细胞毒性淋巴细胞。系统性红斑狼疮(SLE)和混合型 结缔组织病(MCTD)患者产生定向自身抗体 针对不同的抗原部位(分别为Sm和Rnp) 普遍存在的U1小核糖核蛋白颗粒(U1snRNP或U1RNP) 真核细胞。不管是自身抗体还是这个 抗原性U1RNP在这些疾病的发生发展中所起的作用尚不清楚。一个 可靠的抗原性材料来源可能有助于这些调查； 初步数据确定了抗RNP、抗Sm和抗m3G“帽子” 反应性表明原核蓝藻聚球藻 利奥波利斯，可能就是这样的来源。该项目建议将 蓝藻中可检测到的Sm和RNP抗原 它们与人类细胞U1RNP抗原的关系。三甲基1“封顶” 蓝藻的RNA将通过直接RNA和/或cDNA测序进行测序 与已知小RNA序列相关的方法学。 蓝藻提取物的免疫沉淀法和免疫印迹法， 使用表位定义的风湿病患者自身抗体并选择 单抗作为探针，将被用来进一步表征 蓝藻抗原性的蛋白质和核酸组成 材料。竞争性免疫沉淀分析将用于 通过确定人类和蓝藻的成分之间的关系 蓝藻抗原可阻断特定人的免疫沉淀 抗原肽和RNA。细胞形态和细胞内 蓝藻反应性的分布将通过以下方式进行调查 梯度离心法和免疫电子显微镜分析。这些 他们认为研究可以为 理解小核核糖核蛋白的进化起源 粒子(SnRNP)。这样的信息可以阐明SNRNPs的作用 在基本细胞生理学方面发挥作用。

DESCRIPTION: (Adapted from the applicant's abstract) Several rheumatic diseases are associated with the presence of autoimmune antibodies and/or cytotoxic lymphocytes. Both systemic lupus erythematosus (SLE) and mixed connective tissue disease (MCTD) patients produce autoantibodies directed against distinct antigenic sites (Sm and RNP respectively) on the ubiquitous U1 small nuclear ribonucleoprotein particle (U1snRNP or U1RNP) of eucaryotic cells. The role that either the autoantibodies or this antigenic U1RNP plays in the development of these diseases is unknown. A reliable source of antigenic material may facilitate these inquires; preliminary data identifying anti-RNP, anti-Sm, and anti-m3G "cap" reactivity indicate that the procaryotic cyanobacterium, Synechococcus leopoliensis, may be such a source. This project proposes to characterize the Sm and RNP antigens detectable in cyanobacteria in order to determine their relationship to human cell U1RNP antigens. The trimethy1 "capped" cyanobacterial RNA will be sequenced by direct RNA and/or cDNA sequencing methodologies for correlation with known small RNA sequences. Immunoprecipitation assays and western blots of cyanobacterial extracts, using epitope-defined rheumatic disease patient autoantibodies and select monoclonal antibodies as probes, will be used to characterize further the protein and nucleic acid components of the cyanobacterial antigenic material. A competitive immunoprecipitation assay will be used to correlate the human and cyanobacterial components by determining whether cyanobacterial antigens can block the immunoprecipitation of specific human antigenic peptides and RNAs. The cellular form and intracellular distribution of the cyanobacterial reactivity will be investigated by gradient centrifugation and immunoelectron microscopic analyses. These studies they believe could contribute substantial information towards understanding the evolutionary origins of small nuclear ribonucleoprotein particles (snRNPs). Such information could elucidate the role that snRNPs play in basic cellular physiology.