MHC proteins, copolymers and peptide 15mers in EAE

描述（由申请人提供）：人类自身免疫性疾病，其中超过70种是已知的，折磨5-10%的美国人口。一些，如类风湿性关节炎，多发性硬化症和糖尿病发生率高，而其他人，如寻常天疱疮发生率低。所有这些疾病都与特定的组织相容性蛋白有关。本申请的目的是使用多发性硬化症的小鼠模型来研究目前广泛用于治疗这种疾病的药物的改进，并详细研究这些药物起作用的机制。具体而言，我们打算：1.研究新型无规氨基酸共聚物聚（FYAK）n [第二代Copaxone <$1/2]对小鼠实验性自身免疫性脑脊髓炎（EAE）的改善作用（作为其在改善多发性硬化症（MS）中的潜在用途的模型），通过：a）充分表征响应于所述无规氨基酸共聚物聚核苷酸而产生的共聚物特异性、抗原非特异性调节性T细胞系，（F，Y，A，K）n.这些调节性T细胞似乎与已经描述的那些不同，并且它们的产生是共聚物发挥作用的主要机制，B）研究在新模型中通过聚（F，Y，A，K）n改善EAE，c）合成和检查另外的氨基酸共聚物例如聚（V，W，A，K）n和聚（V，Y，A，K）n的性质，以优化在EAE中的功效。2.为了研究具有确定序列的肽15聚体的用途，（第三代Copaxone <$1/2）通过以下方式改善EAE：a）扩展通过J5肽15聚体改善EAE的研究，包括与多聚体的仔细比较，（F，Y，A，K）n和Copaxone在称为疫苗接种、预防和治疗的三种不同方案中的应用; B）类似于上述用氨基酸共聚物的研究，表征用J5肽15聚体免疫后产生的完全调节性T细胞系和克隆; d）合成结合J5肽15聚体的MHC II类四聚体，用其测量组织和外周血中JS特异性调节性T细胞的频率。该方法可以提供一种确定需要施用J5肽15聚体以实现最大治疗功效的频率的手段; e）通过在体内将J5肽15聚体靶向未成熟树突细胞来在体内产生调节性T细胞; f）使用多态性标记物和多光子活体显微术在体内追踪调节性T细胞。__

DESCRIPTION (provided by applicant): Human autoimmune diseases, of which more than 70 are known, afflict 5-10% of the US population. Some, such as rheumatoid arthritis, multiple sclerosis and diabetes occur at high frequency while others, such as pemphigus vulgaris have a low frequency. All of these diseases are linked to specific histocompatibilty proteins. The purpose of this application is to use mouse model of multiple sclerosis to investigate improvements in a drug which is currently in wide use for the therapy of this disease and to study in detail the mechanisms through which these drugs work. In particular we intend: 1. To investigate the use of a novel random amino acid copolymer poly(FYAK)n [a second generation Copaxone¿1/2] for the amelioration of experimental autoimmune encephalomyelitis (EAE) in mice (as a model for their potential use in the amelioration of multiple sclerosis, MS) by: a) fully characterizing copolymer- specific, antigen-non-specific regulatory T cell lines generated in response to the random amino acid copolymer poly(F, Y, A, K)n. These regulatory T cells appear to be different from those already described and their generation is a major mechanism through which the copolymers function, b) investigating the amelioration of EAE by poly(F, Y, A, KJ^n in new models, c) synthesizing and examining the properties of additional amino acid copolymers, for example poly(V, W, A, K)n and poly(V, Y, A, K)n, to optimize efficacy in EAE. 2. To investigate the use of a peptide 15mer of defined sequence (a third generation Copaxone¿1/2) for the amelioration of EAE by: a) extending the studies of amelioration of EAE by the J5 peptide 15mer, including careful comparison with poly(F, Y, A, K)n and Copaxone in three different protocols termed vaccination, prevention and treatment; b) characterizing fully regulatory T cell lines and clones generated after immunization with the J5 peptide 15mer, similarly to the above studies with amino acid copolymers; c) investigating further modifications of the J5 peptide 15mer; d) synthesizing MHC class II tetramers binding the J5 peptide 15mer with which to measure the frequency of JS-specific regulatory T cells tissues and in peripheral blood. This method could provide a means of establishing the frequency with which the J5 peptide 15mer needs to be administered to achieve maximum therapeutic efficacy; e) generating regulatory T cells in vivo by targeting the J5 peptide 15merto immature dendritic cells in vivo; f) Tracking regulatory T cells in vivo using a polymorphic marker and multiphoton intravital microscopy. __