THE RIBONUCLEOPROTEIN AUTOANTIGENS LA AND RO

一个长期存在的医学难题是，为什么一些患有某些风湿病的患者 结缔组织疾病会产生针对自身的抗体 蜂窝组件。对这些疾病发病机制的认识 如果一个人能够批判性地定义结构，疾病就会更容易发生 以及靶抗原的功能。这项提案的主题有两个 这种抗原，La(或SS-B/Sjogren‘s综合征-B)和Ro(或SS-A)。在每一个中 病例中，免疫活性物质已被鉴定为蛋白质。 大自然。这些多肽将被分离出来，并以任何 利用多种技术的独特或共享的物理特性 包括亲和层析、一维和二维凝胶 电泳法和蛋白质印迹法。La和Ro都在 更大的核糖核蛋白复合体的背景，命名为SnRNPs(小的 核RNP)或scRNPs(小细胞质RNPs)，这些将是 利用差示免疫亲和层析进行分离。 每个颗粒都将根据其特有的蛋白质部分进行鉴定 特别强调个别法律可能存在的歧视 和Ro SnRNPs(ScRNP)，包含独特的RNA物种(包括 病毒来源)和包含这两者的那些颗粒的描绘 La和Ro多肽。这些实验的最终结果将是 描述离散的、具有特征的实体，这些实体将与 开发更明确和更灵敏的诊断分析方法 这将有助于对这些细胞的功能研究 抗原及其相关的复合体。在后一端，一种RNA 聚合酶III转录因子与La SnRNP相关 这一因素将被描述为它的 与DNA模板以及与活性物质的其他组件的相互作用 转录复合体。最后，抗La和Ro的单抗将 制作以协助这些研究；它们将使用 杂交瘤技术，并通过体内和体内不同的方案实现 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.