CHARACTERIZATION OF IMMUNE RESPONSES TO S-ANTIGEN

可溶性视网膜抗原（S-Ag）的特征之一是其 能够在眼睛中诱导强烈的自身免疫性炎症， 当在佐剂存在下注射时， 这种疾病称为实验性自身免疫性葡萄膜炎（EAU）， 严重依赖于T细胞和抗原处理， 抗原呈递细胞（APC）。 在1992财政年度， 人S-Ag在刘易斯中的免疫原性和致病性位点 大鼠，使用重叠片段的S-Ag，从而确定， 是S-Ag分子中的几个免疫原性位点，以及至少 三个免疫致病位点 在这些免疫致病位点中，只有 一个是免疫显性的~A片段，位于序列341-360。 的 刘易斯大鼠的免疫原性位点与 在人类中识别。 虽然大多数被认可的网站 大鼠免疫系统聚集在靠近C-末端的地方， 分子，在人体内，大部分活性位于 N-末端区域。 我们还测试了 免疫显性序列，以及非显性序列 免疫致病序列在几个其他大鼠品系中的一个 试图确定是否主要因素 组织相容性复合体（MHC）II类以及MHC外类 II可能在建立免疫原性和免疫原性中发挥作用。 免疫病理反应 在人体实验中，我们测试了建立细胞的新方法， 通过接种多个威尔斯孔的S-Ag致敏外周血 细胞并在培养基中培养14天 富含T细胞生长因子 这种方法产生的数字很低 然后可以通过额外的轮次进一步扩增细胞系 的刺激。 此外，这种方法使我们能够确定 外周血中S抗原反应细胞的前体频率 患者和对照组。 这个前兆频率， 患者中1/1000万循环细胞的数量级， 对照 使用该技术建立的细胞系现在将被 进一步的特点是他们的反应片段， 人类S-Ag以及它们T细胞受体的使用。

One of the characteristics of soluble retinal antigen (S-Ag) is its ability to induce an intense autoimmune inflammation in the eyes of experimental animals when injected in the presence of an adjuvant. This disease, called experimental autoimmune uveitis (EAU), is critically dependent on T cells and antigen processing by appropriate antigen-presenting cells (APC). In FY 1992, we further characterized the immunogenic and pathogenic sites of human S-Ag in the Lewis rat, using overlapping fragments of S-Ag, thus determining that there are several immunogenic sites in the S-Ag molecule as well as at least three immunopathogenic sites. Of these immunopathogenic sites, only one is immunodominant~a fragment located at sequence 341-360. The immunogenic sites for the Lewis rat do not correspond to the sites identified in the human. While most of the sites recognized by the rat immune system are clustered close to the C-terminal end of the molecule, in the human most of the activity is located in the N-terminal region. We also have tested the response of the immunodominant sequence, as well as the nondominant immunopathogenic sequences in several other rat strains in an attempt to determine whether factors within the major histocompatibility complex (MHC) class II as well as outside MHC class II may play a role in the establishment of an immunogenic and an immunopathogenic response. In human subjects, we have tested new means of establishing cell lines by seeding multiple wells with S-Ag primed peripheral blood cells and maintaining the cells in culture for 14 days in medium enriched in T-cell growth factors. This approach yields a low number of cell lines which can then be further expanded by additional rounds of stimulation. In addition, this approach allows us to determine the precursor frequency of cells reactive to S-Ag in the peripheral blood of patients and controls. This precursor frequency, which is of the order of 1 in 10 million circulating cells in patients, is lower in controls. The cell lines established using this technique will now be further characterized in terms of their response to fragments of human S-Ag as well as their T-cell receptor usage.