输血相关急性肺损伤(TRALI)是导致输血相关性死亡的首要原因。有研究显示HLA-A2抗体（A2抗体）与TRALI发生密切相关，但机制不明。我们前期研究发现A2抗体能上调嗜中性粒细胞（PMN）粘附特性和炎性因子分泌及胞内钙信号，而现有研究已证实其有赖于PMN胞内Src磷酸化，提示A2抗体可能通过Src-钙信号通路诱发TRALI。本项目在前期基础上借助Src、Ca2+关键信号分子抑制剂，结合流动小室、荧光实时记录处理手段探讨不同条件下PMN粘附特性和炎性因子分泌的变化；采用荧光标记、细胞培养和流式细胞技术探寻不同条件下PMN钙信号、ROS与肺血管内皮细胞凋亡率的变化规律，以阐明A2抗体通过上调PMN粘附特性和炎性因子分泌及Src-钙信号通路增强钙响应信号而促进ROS产生，介导肺血管内皮细胞损伤诱发TRALI的发病机制，为以A2抗体为靶点防治及发病风险预警和病情监测应用奠定实验基础和理论依据。

Transfusion-related acute lung injury (TRALI) is the leading cause of transfusion-related death. Studies have shown that anti-HLA-A2 is closely related to TRALI occurrence, but the mechanism is unclear. Our previous study have showed that anti-HLA-A2 can raise polymorphonuclear neutrophils (PMN) adhesion properties, inflammatory cytokine secretion and intracellular calcium signaling. It is also reported that activation of PMN immune function are dependent on Src phosphorylation, suggesting that anti-HLA-A2 may activate PMN to induce TRALI through Src-Calcium signal pathway. Based on previous study, we plan to compare the secretion of adhesion characteristics and inflammatory cytokines of PMN with different treatment by combining inhibitors of key signaling molecules (Src or intracellular Ca2+) with flow chamber and real-time fluorescence technique, and also intend to compare calcium signaling and ROS of PMN with different treatment, and the apoptosis rate of pulmonary vascular endothelial cells by fluorescent labeling, cell culture and flow cytometry to clarify the pathogenesis that anti-HLA-A2 induced TRALI through upregulating PMN adhesion properties and inflammatory cytokine secretion and enhancing calcium response signal and ROS production by intracellular Src-Calcium signaling pathway to mediate pulmonary vascular endothelial cell injury, which to lay the theoretical foundation for the possible applications of the prevention of TRALI by targeting to anti-HLA-A2 and in the early warning and monitoring of TRALI.