绿脓杆菌外毒素PEA抗原性高副作用强而临床效用差。前期分析改造获低过敏原性突变体mPEA，IL-5、IL-13产率极显著降低并与空白对照持平，IL-4、IL-6产率虽显著降低却显著高于对照。故认为：内吞入上皮细胞后，mPEA中密集的芳香族残基/亮氨酸残基启动多环芳香烃受体AHR通路、激活NLRC4炎症小体而导致残存的超敏反应。本研究以mPEA为基本模板及对照，合成两类残基不同配置的短肽、免疫刺激性DNA装载无超敏反应的纳米材料ns(mPEA)为试材，刺激细胞系16HBE和THP-1及条件敲除小鼠，LightSheet记录细胞命运，检测CYP1A1、IL-1β、Caspase-1等指标，确证AHR、NLRC4两类炎症事件发生及与特定肽的关联；构建特异HLA基因型的人源化小鼠，基于多个成对材料发掘其间的级联通路，揭示mPEA诱导残存超敏反应的机制，为变态反应哮喘的治疗提供新靶点及新材料。

The clinical application of Pseudomonas aeruginosa exotoxin A (PEA) has been studied for several decades, with the clinical efficacy debated by its high allergenicity. Our previous work showed that the mutants (mPEA), obtained through amino acid substitutions which reduces the HLA binding affinity, induced a significantly lower level of IL-5 and IL-13 compared to wtPEA, while that of IL-4 and IL-6 also declined but still significantly exceeded the blank control. We therefore hypothesize that the residual hypersensitivity may have resulted from the densely populated aromatic amino acid residues and/or leucine residues in certain segments of mPEA endocytosed by epithelial and/or dendritic cells, through activation of the aryl hydrocarbon receptor (AHR) pathway and the NLRC4 inflammasome. In this study, mPEA will be used as the basic template and control. Two types of short peptides composed of different aromatic amino acid and/or leucine residues, and ns(mPEA), nanoscale particles assembled by immunostimulatory DNAs and mPEA, will be used along with their counterpart controls to stimulate the cell lines 16HBE and THP-1 and conditional knock-out mice. The cellular responses will be monitored in real-time by LightSheet Fluorescence Microscopy, accompanied by the investigation of the indicators including CRYP1A1, IL-1β, and caspase-1, etc., in order to confirm the involvement of these two inflammatory events, relating to both AHR and NLRC4 inflammasome, and the causality between the specific peptide(s) and the events. Humanized mice with specific HLA genotypes will be created by adoptively transferring urine stem cells from specific allergic patients to investigate any crosstalk between these two pathways. In conclusion, this study aims to uncover the mechanism by which mPEA induces the residual hypersensitivity, and thus will provide new therapeutic targets and agents for the treatment of allergy.