新型过敏原特异性嗜碱性粒细胞激发试验在过敏诊断中的临床意义研究

Nowadays, serum specific IgE test almost becomes the only laboratory diagnosing test for allergy. The objective fact that allergen specific challenge test is the "golden standard" of allergy diagnosis seems to be completely neglected. Recently, we have developed an allergen specific basophil activation test (BAT) method, and hope to apply clinically to the diagnosis of allergic diseases which should have huge clinical needs from patients. We will use flowcytometry analysis, RNA interference, real time PCR techniques, and our unique aeroallergen specific IgG test kit, skin patch tests for aeroallergens and food allergens, and the second generation of standardized allergen preparations to investigate ① correlations between expression levels of CD203c, FcεRI, CD63 in blood basophils and changes in serum levels of specific IgE and IgG, and results of skin patch tests and skin prick tests; ② expression levels of the above molecules pre and post allergen challenge, the relationship between the levels of CD203c, FcεRI, CD63 and histamine, and the potential mechanisms behind these changes; ③ changes in the levels of CD203c、FcεRI、CD63 on mast cells in nasal lavage fluid and its correlation with expression of the above molecules on blood basophils for a total of 300 patients. Through these investigations, we shall evaluate clinical usage of our novel allergen specific BAT in order to work out a complete laboratory diagnosing procedure for allergic diseases.

目前，血清中特异性IgE水平的检测几乎成了过敏性疾病的唯一诊断方法，忽略了过敏性疾病诊断的"金标准"是过敏原特异性激发试验这一客观事实。本课题组最近研发出了过敏原诱导嗜碱性粒细胞（Bas）激发试验（BAT）方法，有望率先解决过敏性疾病实验室诊断这一有巨大需求的临床问题。本项目应用流式细胞术、RNA干扰、定量PCR等技术，及特有的气传过敏原特异性IgG检测试剂盒，气传过敏原、食物过敏原等斑贴试验试剂盒，第二代标准化过敏原制剂，通过观察300例患者①血液Bas上CD203c、FcεRI、CD63表达变化与血浆特异性IgE、IgG水平变化，皮肤斑贴、点刺试验结果的相关性；②过敏原激发前后上述分子表达变化与组胺分泌的关系及其机制；③过敏性鼻炎鼻灌洗液肥大细胞及血液Bas上述分子表达变化的相关性；临床上系统评价过敏原特异性BAT这一新型技术在过敏诊断中的意义，进一步完善过敏性疾病的实验诊断程序。

目前，血清中特异性IgE 水平的检测几乎成了过敏性疾病的唯一诊断方法，忽略了过敏性疾病诊断的“金标准”是过敏原特异性激发试验这一客观事实。本课题组研发出了过敏原特异性嗜碱性粒细胞（Bas）激发试验（BAT）方法，率先探索解决过敏性疾病实验室诊断这一有巨大需求的临床问题。本项目临床观察了1000 例以上过敏患者，在掌握了过敏患者血液中不同标记的嗜碱性粒细胞群上CD203c、Fcε RI、CD63 表达变化规律的基础上，又发现了新的嗜碱性粒细胞亚型，并且初步发现此亚型能够更好的反映过敏时嗜碱性粒细胞的变化，为实现过敏性疾病的实验诊断提供了新的依据。.课题研究成果包括发表论文28篇（SCI收录论文15篇，中文核心期刊论文13篇）；获省级科技进步二等奖2项；申请并获国家发明专利授权3项；培养研究生6名；主办和参加国内外学术交流7次；并制备出具有全部自主知识产权的过敏原特异性嗜碱性粒细胞激发试验试剂盒，目前已经完成千余例患者的临床检测。