淡水鱼主要致敏原经小鼠肠道树突状细胞CD80/86介导CD4+T细胞活化的机制研究

项目名称： 淡水鱼主要致敏原经小鼠肠道树突状细胞CD80/86介导CD4+T细胞活化的机制研究

项目编号： No.81502810

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 周催

作者单位： 首都医科大学

项目金额： 18万元

中文摘要： 淡水鱼过敏对大众健康的影响已成为当前备受关注的公共卫生问题之一。本课题组前期研究表明致敏模型小鼠肠道中高表达CD80/86的成熟树突状细胞（DCs）数量增多，而DCs及CD80/86分子在鱼类致敏中的作用机制尚不清楚。本研究通过收集淡水鱼过敏人体血清，利用免疫印迹鉴定、分离得到常食用淡水鱼中的主要致敏蛋白，并用其致敏处理C3H/HeJ小鼠后，测定其血清特异性抗体、细胞因子及血浆组胺水平，建立并验证淡水鱼致敏蛋白小鼠致敏模型；通过磁珠分选、小分子干扰RNA和流式细胞术从致敏小鼠获得低表达CD80或CD86的DCs，将其分别与鱼致敏蛋白和CD4+ T细胞共培养，检测T细胞亚群及细胞因子IL-4、IL-5和IL-10的水平，研究肠道DCs亚群通过CD80和CD86活化CD4+ T细胞的机制。本研究对阐明鱼致敏蛋白肠道致敏分子机制有重要意义，可为预防食物过敏提供科学依据。

中文关键词： 淡水鱼；肠道致敏；CD4+；T细胞；树突状细胞；CD80/86分子

英文摘要： Food allergy caused by freshwater fish has been one of the current public health problems because its serious influence to human health. Our preliminary study suggested that number of intestinal dendritic cells (DCs) which high expressing CD80/86 was significantly increased in sensitized mice, while the detailed role and mechanism of DCs and CD80/86 in CD4 + T cell activation has not yet been elucidated. In present study, protein extracts of widely consumed freshwater fish and fish allergic human serum were collected to identify the main allergic proteins in freshwater fish using western blot. Then an ideal mice allergy model to freshwater fish allergen will be established through measuring levels of specific antibodies and cytokines in serum, histamine in plasma and investigate allergic manifestation such as body temperature and blood pressure changes after oral administration.to C3H/HeJ mice. The DCs that low expressing CD80 or CD86 will be obtained by magnetic cell sorting, small interfering RNA and flow cytometry. Then the two kinds of DCs will be co-cultured with freshwater fish allergen and CD4+ T cells respectively. The levels of T cells subsets and cytokines (IL-4, IL-5 and IL-10) will be detected to research the mechanism of CD4+ T cells activation by different DCs subsets through CD80 or CD86 after 96h of co-culture. The results of this research would help us to further clarify the molecular mechanism of intestinal sensitization by fish allergen. Results from the present study would be helpful in prevention of food allergy.

英文关键词： freshwater fish; intestinal sensitization;CD4+ T cell;dendritic cells;CD80/86