Autoimmune Determinants of Human Cardiac Myosin

描述(申请人提供)：心肌炎和风湿性心脏炎 病毒和细菌感染的后遗症分别发生在人类身上 和柯萨奇病毒感染后的动物模型，A组链球菌 感染，或心肌肌球蛋白免疫。这些疾病的发病机制 疾病可能是由于感染性病原体和 宿主自身抗原心肌肌球蛋白。尽管心肌肌球蛋白可以诱导 动物心肌炎，人类心肌炎的分子发病机制是 不清楚。此外，在人类身上很少有研究定义 疾病的免疫学参数。拟议工作的目标是定义 人和人心肌肌球蛋白自身免疫反应的参数 在心肌肌球蛋白诱导的心肌炎和瓣膜炎大鼠模型中。我们会 检验分子模仿的假设和细胞因子的影响 环境导致疾病的发展。我们计划：1)生产和销售 研究心肌炎患者的人类T细胞克隆 与人心肌肌球蛋白、链球菌M蛋白和 柯萨奇病毒及其多肽；检测人T细胞因子应答 细胞克隆以及人类白细胞抗原I、II类反应和细胞的限制 用流式细胞仪分析表面抗原2)评估基因的表达 交叉反应性CD4/-和CD8 T细胞克隆及正常人和心肌炎 患者外周血中CD4和CD8淋巴细胞对刺激的反应 肌球蛋白、M蛋白和柯萨奇病毒及其多肽的DNA阵列 分析，细胞因子的产生，并研究T细胞表面标志物 流式细胞仪分析淋巴细胞3)研究Lewis大鼠心脏S2 多肽诱导的重症心肌炎模型炎症参数的研究 包括分子拟态和细胞因子在心脏病中的作用 易感、耐药和耐受的大鼠品系(Lewis和BB/DR)， 确定心脏特异性浸润性T细胞的表位特异性 从Lewis大鼠模型的心脏中提取细胞，并测定细胞因子在 应用核糖核酸酶保护试验检测心脏TH1/TH2细胞因子及其表达 使用DNA阵列的一大组基因。我们将建立一个宽容模型来 研究肌球蛋白特异性I细胞、抗体和 疾病中的细胞因子。

DESCRIPTION (provided by applicant): Myocarditis and rheumatic carditis are sequelae of viral and bacterial infections, respectively, and occur in humans and animal models following coxsackievirus infection, group A streptococcal infection, or immunization with cardiac myosin. The pathogenesis of these diseases may be due to molecular mimicry between the infectious pathogen and the host autoantigen cardiac myosin. Although cardiac myosin can induce myocarditis in animals, the molecular pathogenesis of the disease in humans is unclear. In addition, there are few studies in humans, which define the immunological parameters of disease. The goal of the proposed work is to define the parameters of the autoimmune response to human cardiac myosin in humans and in a cardiac myosin-induced rat model of myocarditis and valvulitis. We will test the hypothesis that molecular mimicry and the influence of the cytokine environment leads to development of disease. We plan: 1)To produce and investigate human T cell clones from myocarditis patients which are crossreactive with human cardiac myosin, streptococcal M protein and coxsackievirus and their peptides; to determine cytokine responses of human T cell clones as well as HLA class I and II restriction of responses and cell surface antigens by FACS analysis 2)To evaluate expression of genes by crossreactive CD4+/- and CD8+ T cell clones and by normal and myocarditis patient peripheral blood CD4+ and CD8+ lymphocytes in response to stimulation with myosin, M protein and coxsackievirus or their peptides by DNA array analysis, cytokine production, and to study the cell surface markers on T lymphocytes by FACS analysis 3) To investigate a Lewis rat cardiac S2 peptide-induced model of severe myocarditis for parameters of inflammatory heart disease including the role of molecular mimicry and cytokines in susceptible, resistant, and tolerized rat strains (Lewis and BB/DR), to determine the epitope specificity of the heart specific infiltrating T cells from hearts in the Lewis rat model and to determine cytokine expression in hearts using the RNase protection assay for TH1/TH2 cytokines and expression of a large group of genes using DNA arrays. We will establish a tolerance model to investigate the downregulation of myosin specific I cells, antibodies and cytokines in disease.