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70多已知，困扰着5-10%的美国人口。一些疾病，如类风湿性关节炎、多发性硬化症和糖尿病，发生频率较高，而其他一些疾病，如寻常型天疱疮，发生频率较低。所有这些疾病都与特定的组织相容蛋白有关。本申请的目的是利用多发性硬化症的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。1.研究一种新的随机氨基酸共聚物Poly(Fyak)n[第二代Copaxone]用于改善小鼠实验性自身免疫性脑脊髓炎(EAE)(作为其在改善多发性硬化症(MS)方面的潜在用途的模型)的使用：a)充分表征随机氨基酸共聚物Poly(F，Y，A，K)n反应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细胞似乎与已描述的不同，它们的产生是共聚物发挥作用的主要机制，B)研究聚(F，Y，A，KJ^n)在新模型中对EAE的改善；c)合成并检测额外的氨基酸共聚物，例如Poly(V，W，A，K)n和Poly(V，Y，A，K)n，以优化EAE的疗效。2.探讨已定义序列的多肽15多聚体(第三代Copaxone 1/2)用于改善EAE的作用，方法是：a)扩展J5多肽15多聚体改善EAE的研究，包括在疫苗、预防和治疗三种不同方案中与聚(F，Y，A，K)n和Copaxone进行仔细比较；b)鉴定J5多肽15多聚体免疫后产生的完全调节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多肽15多聚体的进一步修饰；D)合成与J5肽15聚体结合的MHC II类四聚体，用于检测JS组织和外周血中JS特异性调节性T细胞的频率。这种方法可以提供一种方法来确定获得最大治疗效果所需的J5肽15聚体的使用频率；e)通过靶向J5肽15肽在体内产生调节性T细胞；f)使用多态标记和多光子活体显微镜追踪体内调节性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. __