AUTOIMMUNITY TO NUCLEAR ANTIGENS

自身免疫性疾病，如狼疮、硬皮病、干燥症 综合征和真皮/多发性肌炎有几组独特的 抗核仁和核仁抗原的自身抗体 用作免疫标记物以区分一种疾病和 其他的。许多自身抗原已经被识别出来，并且是 进化上保守的分子，具有重要的细胞 功能包括DNA合成、转录、RNA处理 还有翻译。将要检验的最新假说是 这些疾病的免疫反应是由抗原驱动的 抗原(S)是一种更大的免疫原性的成分 粒子。第二个假说与地球的守恒有关 该粒子的表位和功能是自身免疫原性 区域(空气)可以是粒子的活性位置，因此 对功能做出贡献。 结合免疫电子显微镜(免疫EM)和 荧光显微镜(LM)将被用来检测核 细胞新陈代谢的不同窗口中的抗原来确定一个 一组自身抗原，如硬皮病中的抗原，可以与 在细胞中的某些点上定位于公共结构区域 新陈代谢。将使用双标记胶体金免疫EM 不同大小的金颗粒，靶子将是细胞 哪些是由化学物质和其他物质引起的功能干扰 操纵与结构变化有关。CDNA克隆 SS-B/La、DNA拓扑异构酶I和34kD纤维蛋白的编码 通过对cDNA3的抗体筛选，可产生蛋白质 表达文库或合成寡核苷酸杂交法。 这些克隆的限制性片段将被插入到 产生了表达载体和亚克隆。融合蛋白 将对这些亚克隆的反应性进行分析 人类自身抗体鉴定序列(S)包含 装腔作势。从序列数据来看，短长度多肽将是 合成并再次检查与自身抗体的反应性 以更严密地界定空气的边界。这 与亲水性简档有关的信息 从该cdna序列推导出的抗原将用于 确定蛋白质表面是否有特殊特征 与自身抗原性有关。最后，抗空气和抗病毒抗体 作为对照的非空气多肽将进行测试，以确定 空气在各自原住民功能中的重要性 蛋白质。

In autoimmune diseases such as lupus, scleroderma, Sjogren's syndrome and dermato/polymyositis there are distinctive sets of autoantibodies to nuclear and nucleolar antigens which can be used as immunological markers to separate one disease from the other. Many of the autoantigens have been identified and are evolutionarily conserved molecules which have important cellular functions including DNA synthesis, transcription, RNA processing and translation. The newest hypothesis which will be examined is that immune responses in these diseaseS are antigen-driven with the antigen(s) being a component of a larger immunogenic particle. The second hypothesis related to the conservation of the epitope and function of the particle is that the autoimmunogenic region (AIR) may be an active site of the particle and therefore contributing to function. A combination of immunoelectron microscopy (immuno EM) and fluorescence light microscopy (LM) will be used to detect nuclear antigens in different windows of cell metabolism to determine if a set of autoantigens such as that in scleroderma can be co- localized in common structural regions at some point in cell metabolism. Double-label colloidal gold immuno EM will be used with different sized gold particles and the targets will be cells in which interference with function induced by chemical and other manipulations is associated with structural changes. cDNA clones encoding SS-B/La, DNA topoisomerase I and 34 KD fibrillarin protein will be generated by antibody screening of cDNA expression libraries or by synthetic oligonucleotide hybridization. Restriction fragments of these clones will be inserted into expression plasmids and subclones generated. The fusion proteins of these subclones will be analyzed for their reactivity with human autoantibodies to identify the sequence(s) containing the AIRs. From the sequence data, short length polypeptides will be synthesized and again examined for reactivity with autoantibodies to more closely define the boundaries of the AIRs. This information in conjunction with the hydrophilicity profile of the antigen deduced from the cDNA sequence will be used to determine if there are special features of the protein surface related to auto-antigenicity. Finally, antibodies to AIR and to non-AIR peptides as controls will be tested to determine the importance of the AIR in the function of their respective native proteins.