S-ANTIGEN--STRUCTURE/FUNCTION RELATIONSHIPS IN UVEITIS

S抗原是一种具有良好特性的视网膜蛋白，与 与视觉过程有关，并且对 诱导实验性自身免疫性葡萄膜炎(EAU)，以及 密切参与视觉的光传递。为了 更加充分地认识S的结构/功能关系-- 抗原在EAU发病机制和光转导中的作用 我们已经制造了(1)单抗(MAb) 它们定义了S抗原的不同表位，(2)确定了 牛S抗原的氨基酸序列，(3)定位的一株单抗 和(4)牛葡萄膜致病部位的定位。 S-抗原。 在我们的实验室中，我们已经产生了两种单抗，MAbA9-C6和MAbA9-C6 MAbA1-G5，它定义了S抗原的不同表位。这些 单抗在我们对S抗原的研究中是有用的 发育中的视网膜、松果体和由此产生的肿瘤 纸巾。牛S抗原的氨基酸序列现已完成 已经确定了。对我们的序列数据的分析显示 S抗原主要以β-片状构象存在； 包含两个潜在的磷酸化位点；三个潜在的 糖类结合部位；ADP-核糖化部位；以及 与视觉中的另一种蛋白质转导蛋白的序列同源性 路径。这些信息对于理解 S抗原在视觉光传导中的作用。一种知识 氨基酸序列的同源性也导致了对 MAbA9-C6结合部位和两个葡萄膜致病部位。 与S抗原的氨基酸序列相对应的多肽， 已被化学合成，并指定了两个多肽 多肽K和多肽M被发现高度 葡萄膜致病。微克疫苗免疫Lewis大鼠的研究 两种多肽中的任何一种都能引起临床上的EAU。 在组织病理学上与由以下因素引起的EAU没有区别 土生土长的S-抗原。 在这份续订方案中，我们计划完善和扩展我们的 关于牛葡萄膜致病部位的初步观察， 人和大鼠S抗原。此外，我们建议生产 抗葡萄膜致病决定因素的单抗和提纯 MAbA9-C6结合部位要分解为单一氨基酸。 这些研究和我们正在开发的试剂不仅有助于 为了了解S抗原在EAU发病机制中的作用， 在视觉的光传输方面也是如此。

S-antigen is a well-characterized retinal protein, intimately involved in the visual process, and highly pathogenic for the induction of experimental autoimmune uveitis (EAU), and intimately involved in the phototransduction of vision. In order to more fully understand structure/function relationships of S- antigen in the pathogenesis of EAU and in the phototransduction of vision we have (1) produced monoclonal antibodies (MAbs) which define different epitopes of S-antigen, (2) determined the amino acid sequence of bovine S-antigen, (3) localized one MAb binding site and (4) localized two uveitopathogenic sites in bovine S-antigen. In our laboratory, we have generated two MAbs, MAbA9-C6 and MAbA1-G5, which define different epitopes of S-antigens. These MAbs have been useful in our studies concerning S-antigen in the developing retina, pineal gland and in tumors arising from these tissues. The amino acid sequence of bovine S-antigen has now been determined. Analysis of our sequence data has revealed that; S-antigen exists primarily in a beta sheet conformation; contains two potential phorphorylation sites; three potential carbohydrate attachments sites; an ADP-ribosylation site; and sequence homologies to tranducin, another protein in the visual pathway. Such information is essential in order to understand the role of S-antigen in the phototransduction of vision. A knowledge of the amino acid sequence has also led to the identification of the MAbA9-C6 binding site and two uveitopathogenic sites. Peptides, corresponding to the amino acid sequence of S-antigen, have been synthesized chemically and two peptides designated peptide K and peptide M have been found to be highly uveitopathogenic. Immunization of Lewis rats with microgram amounts of either peptides induced an EAU which was clinically and histopathologically indistinguishable from the EAU induced by native S-antigen. In this continuation proposal we plan to refine and expand our initial observations regarding the uveitopathogenic sites in bovine, human and rat S-antigen. In addition, we propose to produce MAbs against the uveitopathogenic determinants and to refine the MAbA9-C6 binding site to the resolution of a single amino acid. These studies and the reagents we are developing will not only aid in understanding the role of S-antigen in the pathogenesis of EAU, but in the phototransduction of vision as well.