脓毒症时中性粒细胞经P38/MSK-1/MK-2途径抑制T细胞免疫功能的机制研究

脓毒症时中性粒细胞生物学功能异常和免疫抑制是导致机体免疫系统失衡、器官功能不全的重要因素。我们前期研究显示，脓毒症时中性粒细胞相关免疫抑制基因和蛋白表达异常，尤其对T细胞有明显的抑制效应，但详细机制不清。本研究采用基因敲除动物模型、CRISPR-Cas9基因编辑细胞模型，运用单细胞测序、蛋白质定位定量测定等手段，在分子、细胞和整体等不同层面上，研究脓毒症时中性粒细胞相关免疫抑制基因和蛋白的动态变化、对T细胞活化和转分化的调节作用、P38/MSK-1/MK-2信号途径的调控作用及机制，进一步利用原子力显微镜平台和多色免疫荧光二维重建技术精准测定脓毒症时中性粒细胞与T细胞之间的空间相关关系，旨在探明脓毒症时中性粒细胞对T细胞活化和转分化调节的重要特征，详细阐明P38/MSK-1/MK-2信号途径调控中性粒细胞表现免疫抑制的（部分）分子学基础，为发现脓毒症干预新靶点提供新思路。

Abnormal biological functions and immunosuppressive characteristics of neutrophil in sepsis are important factors leading to imbalance of immune system and organ dysfunction. Our previous studies have revealed that expressions of immunosuppressive genes and proteins of neutrophil in sepsis are upregulated, the detail mechanisms, however, are not completely elucidate. This project employs gene knock-out animal models and CRISPR-Cas9 gene editing system to examine the dynamic state of expressions of immunosuppressive genes and proteins of neutrophil during sepsis. The important factors will be investigated in the molecular and cellular levels through applying updated molecular biological techniques including gene analysis, 10X Genomics single cell sequencing、mass spectrometric analysis, measurement of protein expressions, which investigate the dynamic state of neutrophil functions, the dynamic changes of T cell activation, expressions of related immunosuppressive genes and proteins during sepsis. Furthermore, this project also focuses on the important influences of the adjustment disorder of P38/MSK-1/MK-2 signal transduction in neutrophil to immune functions. Therefore, we are in attempt to elucidate the disorder of neutrophil regulating capacity during sepsis to determine whether P38/MSK-1/MK-2 signal transduction will be the novel targets, in order to elucidate molecular mechnisms (even partly) responsible for the immunoregulation of neutrophil and to provide theoretical basis and new idea for the research and treatment of sepsis.