PS通过TIM-1激活Src介导T淋巴细胞活化及其在TRALI发生发展中的作用机制

TRALI是导致输血相关性死亡的首要的严重不良反应，目前缺乏有效治疗手段。研究表明血液制备、储存所产生的PS是诱发TRALI的重要非特异性免疫因素，但作用机制不清。我们前期发现沉默PS受体TIM-1或抑制Src活性可抑制PS活化T细胞介导TRALI的发生，推测PS通过TIM-1激活Src活化T细胞是诱发TRALI的关键通路。本研究拟在已有工作基础上，通过CO-IP、定点突变和基因沉默等技术探讨PS活化Src过程中TIM-1与Src相互作用；运用流式细胞、测序和质谱等技术探索Src调控的下游信号通路；采用流动小室、双荧光图像处理等技术探讨PS诱导T细胞胞内钙响应、骨架重排和粘附特性及免疫效应的变化；利用T细胞特异性条件敲除TIM-1小鼠模型和Src活性抑制进一步阐明PS通过TIM-1激活Src活化T细胞诱发TRALI的发生发展，为PS诱发TRALI的疾病预防及靶向治疗提供实验基础和理论依据。

TRALI is the first serious adverse reaction which leads to transfusion related death, and there is no effective treatment at present. Studies have shown that PS produced during blood preparation and storage is an important non-specific immune factor for TRALI, but the mechanism is unclear. We have found that silencing TIM-1 or inhibiting the activity of Src kinase could inhibit T lymphocyte activity and TRALI induced by PS. It is suggested that the activation of Src mediates by PS combined with TIM-1 is the key pathway which leads to TRALI induced by T lymphocyte activation. In this study, based on the previous work, the interaction between TIM-1 and Src and its mechanism in PS activated Src were studied by CO-IP, site directed mutation and gene silencing. Flow cytometry, sequencing and mass spectrometry were used to analyze the changes of molecular signal pathway after Src activation. Flow chamber and double fluorescence were used to observe the calcium response, cytoskeleton rearrangement, adhesion level and immune effection of T lymphocyte mediated by PS. The mouse model of T lymphocyte specific TIM-1 knockout and Src inhibitor were used to further verify that PS activated Src through TIM-1 which mediated T lymphocyte activation and its role in the development of TRALI. It provides experimental basis and theoretical basis for the disease prevention and targeted treatment of TRALI induced by PS.