绘制尘螨过敏原全谱并揭示其致敏机制是变态反应学基础性科学目标。课题组虽首次解析了粉尘螨基因组、转录组和微生物组图谱；并发现和命名了主要过敏原Der f24（JACI，2015），但尚未建立高通量过敏原组学技术，Der f24致敏机制尚不明确。为此，继续研究并获得：① Der f24 IgE主要线性表位位于N端1-32aa；②Der f24促进上皮细胞分泌TSLP，使NFAT去磷酸化，并被钙离子通道抑制剂抑制。在此基础上，开展抗IgE偶联亲和层析分离纯化与高分辨LC-MS/MS结合、肽阵列、点突变、信号通路分析等实验，拟解决科学问题：①建立高通量过敏原组学分析技术，解析粉尘螨IgE反应蛋白质组；②解析Derf24 关键IgE线性表位；③验证Der f24通过STIM1/ORAI1/NFAT通路促进TSLP致敏的假说。本项目有望克服过敏原组学高通量技术瓶颈，为过敏性疾病防治奠定基础。

Elucidation of the full spectrum of dust mite allergens and their immunological mechanism are key allergological objectives. Through continuous efforts ，we published a description of the dust mite genome, transcriptome and microbiome and discovered a new major dust mite allergen, Der f 24（JACI，2015，135(2):539-548）. We have obtained clues as to Der f 24’s immunological mechanism, including our observations that Der f 24 promotes epithelial cell secretion of thymic stromal lymphopoietin (TSLP), possible through dephosphorylation of nuclear factor of activated T cells (NFAT), and that this effect is attenuated by calcium ion channel inhibitors. Additionally, we have data indicating that the major IgE linear epitopes of Der f 24 are near the N-terminus (amino acids [aa] 1–32). Important allergologic problems remain that could be tackled by the establishment of a high-throughput allergenomic technology platform. Hence, building on our experience, our plan is to move forward with the following three aims: (1) establish a high-throughput allergenomic technology that enables all IgE-reactive proteins to be identified in a single pass (2) analyze the main IgE linear epitopes of Der f 24; and (3) test the hypothesis that Der f 24 promotes TSLP secretion via the stromal interaction molecule 1 (STIM1)/ Calcium release-activated calcium channel protein 1 (Orai1) /NFAT pathway. We will employ multiple complementary methods including anti-human IgE binding affinity chromatography, liquid chromatography-tandem mass spectrometry (LC-MS/MS), peptide arrays, mutation studies, and signaling pathway analysis. Completion of these aims will resolve the bottleneck of high-throughput allergenomic technology and lay the groundwork for identification of the full spectrum of dust mite allergens and the development of diagnostics and immunotherapeutic vaccines for HDM allergic diseases.