过敏原诱导肥大细胞/嗜碱性粒细胞CD203c表达变化的机制及其临床意义研究

Abstract：It has long been recognized that allergen-induced mast cell and basophil degranulation tests are decisive tests for diagnosing allergic disorders in vitro. In recent years, CD203c, a reliable marker for basophil activation has been discovered, which allows us for the first time to have the proper tools to investigate and answer if activation of mast cells and basophils definitely causes degranulation, a long-term unsolved scientific question. In the present study, flowcytometry, RNA interference, real time PCR and other techniques will be employed to study: ①the potential correlations between expression of CD203c and CD123 in whole blood and specific IgE or histamine levels in the plasma of allergic patients; ②correlations of expression of CD117、CD203c、CD123 on nasal mast cells and histamine release from the mast cells pre or post allergen challenge; ③influence of allergens, haptens or low molecular weight activators on expression of CD203c and histamine release from passively sensitized highly purified mast cells and basophils, and its potential mechanisms and ④regulatory actions of cytokines on expression of CD203c and histamine release from mast cells and basophils, and its potential mechanisms, therefore to elucidate the role of CD203c in the pathogenesis of allergic inflammation and its diagnostic value for allergic disorders, and to clarify the relationship between mast cell/basophil activation and degranulation.

摘要：过敏原诱导的肥大细胞(MC)/嗜碱性粒细胞(Bas)脱颗粒试验（测组胺）早已被公认是过敏性疾病体外诊断的定性试验。近年来，人们又发现CD203c这一可靠的Bas激活的标记物，从而使回答是否MC/ Bas激活一定会出现脱颗粒这个十分重要的科学问题有了真正可行性。本项目将应用流式细胞术、RNA干扰、定量PCR等技术，通过观察①过敏患者血液中Bas上CD203c、CD123表达变化与血浆中特异性IgE/组胺水平变化的相关性；②过敏原激发前后过敏患者鼻腔中MC上CD117、CD203c、CD123表达变化与组胺水平变化的相关性；③过敏原、半抗原及小分子激活剂对被动致敏的高纯度MC/Bas的CD203c表达、组胺分泌的影响及机制；④细胞因子对MC/Bas的CD203c表达、组胺分泌的调节作用及机制；揭示CD203c在过敏性炎症中的作用机制和临床诊断意义，以及MC/Bas激活与脱颗粒的关系。

本项目用流式细胞术检测过敏患者血液中嗜碱性粒细胞经进行体外临床可疑过敏原激发前后，血嗜碱性粒细胞CD203c、CD123、HLA-DR、CD63、FcRI、CCR3共6个指标表达变化；用UNIPCAP方法检测过敏患者屋尘、螨虫、蟑螂、杨柳榆树、豚草、葎草、蒿草、鱼、花生、牛奶、黄豆、鸡蛋、霉菌、猫毛特异性IgE水平；用ELISA方法检测过敏患者血清中IL-13、IL-17A、IL-18、IL-28A、IL-29、IL-31、IL-33、TSLP水平表达。发现CD203c+CCR3；CD123+HLA-DR+CD63；CD123+HLA-DR+FcRI 组合均能不同程度的反映嗜碱性粒细胞激活状态（可靠性达90%以上）；但是CD203c+CCR3+CD123+HLA-DR+CD63组合不仅能表达出嗜碱性粒细胞激活状态还能表达出嗜碱性粒细胞脱颗粒状态（可靠性达96%以上）；初步建立起与国际先进水平接轨的过敏原诱导MC/Bas激活和脱颗粒的高技术试验平台；已经开始分析皮肤疾病患者CD203c、CD123、HLA-DR、CD63、FcRI、CCR3表达与血清中IL-13、IL-17A、IL-18、IL-28A、IL-29、IL-31、IL-33、TSLP水平表达、不同过敏原特异性IgE阳性患者间的相关性。本项目研究已发表论文7篇（其中SCI收录论文2篇，中文核心期刊收录论文3篇），已投在审SCI收录论文1篇，正在撰写SCI收录论文1篇（将于2014年发表）。