RIBONUCLEOPROTEIN AUTOANTIGENS LA AND RO

一个长期存在的医学难题是为什么有些患有某些风湿性关节炎的患者 结缔组织疾病会产生抗体 细胞成分。 了解这些疾病的发病机制 如果我们能批判性地定义 和靶抗原的功能。 本提案的主题有两个 这些抗原是La（或SS-B/Sjogren综合征-B）和Ro（或SS-A）。 在每个 例，免疫反应性物质已被确定为蛋白质， 自然 这些多肽将被分离并以任何形式描述。 独特或共享的物理特性， 包括亲和层析、一维和二维凝胶 电泳和蛋白质印迹。 La和Ro都在 命名为snRNP的较大核糖核蛋白复合物（小 核RNP）或scRNP（小细胞质RNP），并且这些将是 分离，通过使用差异免疫亲和层析辅助。 每个颗粒将根据其特征蛋白质部分进行鉴定 特别强调对个人的可能歧视， 和含有独特RNA种类的Ro snRNP（scRNP）（包括 病毒来源）和对那些含有两者的颗粒的描绘 La和Ro多肽。 这些实验的最终结果将是 描述离散的，特征化的实体， 开发比现有技术更明确和更灵敏的诊断检测方法 目前可用，这将有助于这些功能的研究 抗原及其相关复合物。 最后，一种RNA 聚合酶III转录因子与La snRNP相关 这个实验室;这个因素将被描述为 与DNA模板和活性物质的其他组分的相互作用 转录复合体 最后，抗La和Ro单克隆抗体将 制作以协助这些研究;它们将使用 杂交瘤技术并通过体内和 体外免疫。

A long existent medical puzzle is why some patients with certain rhematic and connective tissue diseases produce antibodies against their own cellular components. An understanding of the pathogenesis of these diseases would be facilitated if one could critically define the structure and function of the target antigens. The subjects of this proposal are two such antigens, La (or SS-B/Sjogren's syndrome-B) and Ro (or SS-A). In each case, the immunoreactive species have been identified as protein in nature. These polypeptides will be isolated and described in terms of any unique or shared physical properties utilizing a number of techniques including affinity chromatography, one- and two-dimensional gel electrophoresis and protein blots. Both La and Ro function within the context of larger ribonucleoprotein complexes designated snRNPs (small nuclear RNPs) or scRNPs (small cytoplasmic RNPs) and these will be isolated, aided by the use of differential immunoaffinity chromatography. Each particle will be identified as to its characteristic protein moieties with particular emphasis on the possible discrimination of individual La and Ro snRNPs (scRNPs) that contain unique RNA species (including those of viral origin) and on the delineation of those particles that contain both La and Ro polypeptides. The end result of these experiments will be to describe discrete, characterized entities that willhave relevance in the development of more defined and sensitive diagnostic assays than are currently available, and that will facilitate functional studies of these antigens and their associated complexes. To the latter end, an RNA polymerase III transcription factor has been associated with the La snRNP by this laboratory; this factor will be characterized as to its interactions with DNA templates and with the other components of the active transcription complex. Lastly, anti-La and Ro-monoclonal antibodies will be produced to assist in these studies; they will be made using the hybridoma technology and implemented by different protocols of in vivo and in vitro immunization.