Cytokine Regulation of Collagen-induced Arthritis

描述(由申请人提供)： 胶原性关节炎是一种研究较多的类风湿性关节炎动物模型。在免疫佐剂中的异种II型胶原蛋白诱导炎症反应后，会发生一种自身免疫过程。引起的炎症反应的性质可能与所使用的抗原一样关键，因为以前被认为对疾病具有抵抗力的小鼠品系在免疫方案改变时可能会变得敏感。这项应用的长期目标是开发一项独立的研究计划，在对自身免疫性疾病发生所需的炎症反应进行更清楚的表征的基础上，提高对炎症性关节炎的病因的理解。 首先，将进行免疫调节性Th2细胞因子在疾病进展中的表征。IL-4和IL-10缺陷小鼠将被II型胶原免疫，并将监测关节炎的发展。在缺乏内源性IL-4或IL-10的情况下，将通过细胞因子和趋化因子的表达来研究对胶原的免疫反应。也将研究不同免疫方案导致的T细胞对抗原的反应，因为T细胞谱系的差异可能是疾病存在或不存在的原因。将确定T细胞受体V3基因的使用以及胶原反应性T细胞的表位特异性。将测定胶原反应性抗体水平和结合特异性，以评估对B细胞反应性的影响。最后，将确定对这些免疫方案的先天免疫反应。用流式细胞术和免疫印迹法研究Toll样受体的表达和功能，并测定在不同免疫条件下产生IL-12和IL-18等细胞因子的能力。通过更好地了解疾病发展所需的独特炎症途径，可能会设计出治疗类风湿性关节炎等自身免疫过程的特定免疫治疗策略。

DESCRIPTION (provided by applicant): Collagen-induced arthritis is a well-studied animal model of human rheumatoid arthritis. An autoimmune process evolves after immunization with heterologous type II collagen in an adjuvant that induces an inflammatory response. The nature of the inflammatory response induced may be as critical as the antigen used, for mouse strains that have been previously believed to be resistant to disease can become susceptible when the immunization protocol is modified. The long-term objectives of this application are to develop an independent program of research that leads to an improved understanding of the etiopathogenesis of inflammatory arthritis based on a clearer characterization of the inflammatory responses necessary for an autoimmune disease to develop. Initially, characterization of immunomodulatory Th2 cytokines in disease progression will be performed. IL-4 and IL-10 deficient mice will be immunized with type II collagen, and the development of arthritis will be monitored. The immune response to collagen will be studied as determined by cytokine and chemokine expression in the absence of endogenous IL-4 or IL-10. The T cell response to antigen resulting from different immunization protocols will also be studied, as differences in the T cell repertoire may be responsible for the presence or absence of disease. T cell receptor V3 gene usage as well as epitope specificity of collagen-reactive T cells will be determined. Levels of collagen-reactive antibodies and binding specificities will be determined to assess the effect on B cell reactivity. Finally, innate immune responses to these immunization protocols will be ascertained. The expression and function of toll-like receptors will be studied by flow cytometry and immunoblotting, and the ability to produce cytokines such as IL-12 and IL-18 under different immunization conditions will be determined. By better understanding unique inflammatory pathways that are required for the development of disease, specific immunotherapy strategies for the treatment of autoimmune processes such as rheumatoid arthritis may be designed.