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，其中已知超过70种，折磨着5-10%的美国人口。有些疾病，如风湿性关节炎、多发性硬化症和糖尿病，发生频率很高，而另一些疾病，如寻常性天疱疮，发生频率很低。所有这些疾病都与特定的组织相容性蛋白有关。本申请的目的是利用多发性硬化症小鼠模型来研究目前广泛用于治疗多发性硬化症的药物的改进，并详细研究这些药物的作用机制。我们特别打算：1；为了研究一种新型随机氨基酸共聚物（FYAK）n[第二代Copaxone¿1/2]在改善小鼠实验性自身免疫性脑脊髓炎（EAE）中的应用（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)充分表征响应随机氨基酸共聚物（F， Y, a， K）n产生的共聚物特异性，抗原非特异性调节性T细胞系。这些调节性T细胞似乎与已经描述的不同，它们的产生是共聚物功能的主要机制，b)在新模型中研究聚（F， Y, a， KJ^n对EAE的改善，c）合成和检查其他氨基酸共聚物的性质，例如聚（V， W, a， K）n和聚（V， Y, a， K）n，以优化EAE的功效。2. 研究确定序列的肽15mer（第三代Copaxone¿1/2）用于改善EAE的使用：a)扩展J5肽15mer改善EAE的研究，包括在三种不同的方案中与poly(F， Y, a， K)n和Copaxone进行仔细比较，称为疫苗接种，预防和治疗；b)表征J5肽15mer免疫后产生的完全调节性T细胞系和克隆，类似于上述氨基酸共聚物的研究；c)研究J5肽15mer的进一步修饰；d)合成结合J5肽15mer的MHC II类四聚体，用于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方法，确定J5肽15mer需要施用的频率，以达到最大的治疗效果；e)通过J5肽15merto体内未成熟树突状细胞产生体内调节性T细胞；f)使用多态标记和多光子活体显微镜跟踪体内调节性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. __