IMMUNOGENIC EPITOPES OF COLLAGEN CRITICAL IN ARTHRITIS

对正常关节蛋白的免疫反应可能起着关键作用。 在类风湿性关节炎关节破坏中的作用。许多患者都有 对各种结缔组织成分和关节炎的可检测免疫力 可以通过用胶原蛋白或用来免疫啮齿动物来诱导 从软骨中分离出的蛋白多糖。发现这种抗原 与MHC分子结合并被T细胞识别的片段可以是 以为代表的线性合成肽极大地促进了 阐明抗原识别的确切性质。本地化 结缔组织产生免疫原性的决定因素 这些反应可能会让我们更好地了解关节炎 是启动和监管的。这项提议的目的包括确定 具有免疫原性的II型胶原决定因素(CII)。二 将采取各种方法来确定结构要求 既能抑制关节炎又能诱导关节炎的抗原。多种多样 多肽将由溴化氰和氰化物的组合产生 蛋白水解性切割 II型胶原蛋白，并将被提纯。合成肽(23-25个氨基酸 酸残基长)表示分子的离散区域 准备好了。胶原蛋白碎片和合成肽将用于 在免疫CII前耐受小鼠，并观察随后的 抑制关节炎。碎片还将与CFA和 用于免疫DBA/1小鼠，随后将观察 关节炎的发展。然后T细胞株和克隆与 CII的决定簇将从猪的脾和淋巴结中分离出来 免疫和耐受小鼠，并测试其诱导 对关节炎的耐受和随后的抑制，导致关节炎本身， 或改变既得利益的疾病。最后，我们将确定活动站点 使用重叠多肽和具有以下特性的多肽对每个免疫原肽进行 单一氨基酸的取代和活性测试 用体外T细胞增殖试验和体内试验测定 它们是否会引起与亲本多肽相同的免疫反应。 阐明自身抗原耐受性的机制 诱导和维持对急性淋巴细胞白血病的治疗具有重要意义 自身免疫性疾病。

The immune response to normal joint proteins probably plays a critical role in the joint destruction of rheumatoid arthritis. Many patients have detectable immunity to various connective tissue components and arthritis can be induced in rodents by immunizing them either with collagen or proteoglycans isolated from cartilage. The discovery that the antigen fragments that bind to MHC molecules and are recognized by T cells can be represented by, linear synthetic peptides has greatly facilitated the elucidation of the exact nature of antigen recognition. Localizing determinants of connective tissue with the capacity to generate immunogenic responses may give us a better understanding of the way in which arthritis is initiated and regulated. The aims of this proposal include identifying determinants of type II collagen (CII) which are immunogenic. Two approaches will be taken to identify the structural requirements of antigens mediating both suppression and induction of arthritis. various peptides will be generated by a combination of cyanogen bromide and proteolytic cleavage of type II collagen and will be purified. Synthetic peptides (23-25 amino acid residues long) representing discrete regions of the molecule will be prepared. Collagen fragments and synthetic peptides will be used to tolerize mice prior to immunization to CII and observed for the subsequent suppression of arthritis. Fragments will also be emulsified with CFA and used to immunize DBA/1 mice which will subsequently be observed for the development of arthritis. Then T cell lines and clones reactive with determinants of CII will be isolated from spleens and lymph nodes of immunized and tolerized mice and tested for the ability to either induce tolerance and subsequent suppression of arthritis, induce arthritis itself, or alter established disease. Finally we will determine the active site of each immunogenic peptide using overlapping peptides and peptides having single amino acid substitutions and testing these peptides for activity using in vitro T cells proliferative assays and in vivo assays to determine whether they elicit the same immune response as the parent peptide. Elucidating the mechanisms by which tolerance to autoantigens can be induced and maintained has important implication for therapy of all autoimmune diseases.