IMMUNOGENIC EPITOPES OF COLLAGEN CRITICAL IN ARTHRITIS

对正常关节蛋白的免疫反应可能发挥着关键作用 类风湿性关节炎的治疗方法有哪些 许多患者已 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 从软骨中分离的蛋白聚糖。 发现抗原 结合MHC分子并被T细胞识别的片段可以被 以线性合成肽为代表， 阐明抗原识别的确切性质。 定位 具有产生免疫原性能力的结缔组织决定簇 反应可能会让我们更好地了解关节炎的方式， 它被启动和管理。 该提案的目的包括： II型胶原（CII）的决定簇，其具有免疫原性。 两 将采取各种办法，确定 介导抑制和诱导关节炎的抗原。各种 肽将通过溴化氰和 的蛋白水解切割 II型胶原蛋白，并将被纯化。 合成肽（23 - 25个氨基酸） 酸残基长）代表分子的离散区域， 制备 胶原蛋白片段和合成肽将用于 在对CII免疫之前使小鼠耐受，并观察随后的 抑制关节炎。 片段也将用CFA乳化， 用于免疫DBA/1小鼠，随后将观察其免疫反应。 关节炎的发展。 然后T细胞系和克隆与 CII的决定簇将从脾和淋巴结中分离， 免疫和耐受化的小鼠，并测试诱导 关节炎耐受和随后的抑制，诱发关节炎本身， 或改变已有的疾病。 最后，我们将确定活动站点 每种免疫原性肽使用重叠的肽和具有 单个氨基酸取代并测试这些肽的活性 使用体外T细胞增殖测定和体内测定来确定 它们是否引发与亲本肽相同的免疫应答。 阐明对自身抗原耐受的机制， 诱导和维持对于治疗所有 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.