TISSUE-SPECIFIC ANTIGEN RECEPTOR REPERTOIRES IN RA

严重活动期类风湿性关节炎的病变滑膜包含 脾内能产生免疫球蛋白的淋巴细胞生发中心 级别。对这些免疫球蛋白的研究主要集中在类风湿因子上。 (RF)含量，并表明，在血清阳性的RA中，5%-10%的 曲目包括1GM和1GG抗1GG FC。有很强的 间接证据表明所产生的免疫复合体参与了 疾病的发病机制和进展。然而，这三位 独特型最常见的表达是1gm的RF副蛋白，例如， Wa、Po和Bla似乎是滑膜RF的一个次要成分。因此， 类风湿关节炎滑膜抗体谱系的遗传组成--均为非RF 而射频--仍然是一个悬而未决的问题。在这个项目的过程中，我们 将分析滑膜单个核细胞表达的抗体库 以检验滑膜免疫球蛋白的假说 产生是由抗原驱动的，因此参与了发病机制。 这种疾病的危害。对这种方法的相当大的支持来自于小鼠 模特们。MRL/LPR小鼠抗体库的序列分析 提示在非特异性刺激过程中产生的自身抗体 LP或二次免疫似乎是多克隆的，反映了 生殖系谱系；而在自身免疫状态下，高度自我反应 抗体是克隆相关的，并表现出所有的特征 抗原驱动的免疫球蛋白的产生。小鼠模型也表明 特定的V种系元素的物理化学性质可以 影响自身免疫性疾病的表现，因此多态或 基因组谱系中的异形性可能是一个额外的、独立的 疾病易感性的因素。类风湿滑膜组织中的c DNA文库 将产生细胞，并在 核苷酸序列水平。V区利用受限的证据 在CMU-和Ckappa-包含的成绩单将被寻找。比较将会 在RF副蛋白之间，生殖系隔间，EBV转化 类风湿关节炎患者滑膜细胞株及不同滑膜细胞系 疾病的各个阶段。抗原特异性克隆扩增的证据将是 通过分析具有代表性的CGamma-And的VH-DH-JH连接寻求 含有Calpha的转录本以及Vkappa-Jkappa的cDNA。分析 这些克隆的体细胞突变的程度和分布将 提供支持或反对抗原驱动的选择的额外证据。 将生成组合表达文库，以测试可能的 抗原特异性。候选生殖系自身反应元件将是 它们在人类群体中的多态程度 以及它们与选定疾病的相关性(如果有) 已确立的表现。这些研究不仅应该使重要的 光变成了一种鲜为人知的风湿病的表现，但也 为今后对异常免疫反应的研究提供参考。

Diseased synovium in severe, active rheumatoid arthritis contains lymphocyte germinal centers able to produce immunoglobulin at splenic levels. Studies of these immunoglobulins have focused on rheumatoid factor (RF) content and have shown that, in seropositive RA, 5-10% of the repertoire consists of both 1gM and 1gG anti-1gG Fc. There is strong circumstantial evidence that the resultant immune complexes are involved in the pathogenesis and progression of the disease. However, the three idiotypes which are most commonly expressed by 1gM RF paraproteins, e.g., Wa, Po and Bla, appear to be a minor component of synovia RF. Thus, the genetic composition of the RA synovial antibody repertoire - both non-RF and RF - remains an open question. During the course of this project, we will analyze the antibody repertoire expressed by synovial mononuclear cells in order to test the hypothesis that synovial immunoglobulin production is antigen-driven and, therefore, involved in the pathogenesis of the disease. Considerable support for this approach derives from murine models. Sequence analysis of antibody repertoires in the MRL/lpr mouse suggests that autoantibodies generated during nonspecific stimulation by LPS or secondary immunization appear to be polyclonal and reflect the germline repertoire; whereas in autoimmune states, highly self-reactive antibodies are clonally related and demonstrate all the characteristics of antigen-driven immunoglobulin production. Murine models also suggest that the physico-chemical properties of specific V germline elements may influence the manifestations of autoimmune disease, hence polymorphism or heteromorphism in the genomic repertoire may be an additional, independent factor in disease susceptibility. cDNA libraries from rheumatoid synovial cells will be generated and the antibody repertoire characterized at the nucleotide sequence level. Evidence of restriction in V region utilization in Cmu- and Ckappa-containing transcripts will be sought. Comparisons will be made between RF paraproteins, the germline compartment, EBV-transformed synovial cell lines from RA patients, and synovial repertoires at different stages of disease. Evidence of antigen-specific clonal expansion will be sought by analyzing the Vh-Dh-Jh joins of representative Cgamma- and Calpha-containing transcripts as well as the Vkappa-Jkappa cDNAs. Analysis of the extent and distribution of somatic mutation among these clones will provide additional evidence for or against antigen-driven selection. Combinatorial expression libraries will be generated to test for possible antigen specificity. Candidate germline autoreactive elements will be identified, their extent of polymorphism in the human population established, and their correlation, if any, to selected disease manifestations established. These studies should not only shed significant light into a poorly understood manifestation of rheumatic disease, but also serve as reference for future studies of abnormal immune responses.