PK2调控中性粒细胞功能在铜绿假单胞菌诱导的细菌性肺炎模型中的免疫机制研究

铜绿假单胞菌感染引起的细菌性肺炎已成为威胁人类健康的全球性问题。随着抗生素耐药率的增加，寻找替代性预防和治疗策略是当务之急，而免疫调节因子的应用为临床治疗提供了新方向。课题组前期发现PK2对铜绿假单胞菌诱导的肺炎具有保护作用，可能与增强了中性粒细胞募集和吞噬杀伤功能有关，但具体机制不清。由此提出“PK2可通过激活呼吸道上皮细胞相关信号通路，产生趋化因子，调控中性粒细胞免疫应答，改善肺部免疫稳态，增强宿主抗菌反应，进而发挥保护作用”的科学假说。本研究拟借助缺陷小鼠和体外原代细胞模型，采用荧光定量PCR、免疫荧光定位、流式细胞分选术、基因芯片、Luminex多重蛋白检测以及CRISPR/Cas 9等技术，在分子、细胞和动物三个层次上揭示“PK2调控中性粒细胞免疫应答功能”在铜绿假单胞菌诱导的细菌性肺炎中的作用及分子信号机制，为以PK2为靶点研发新的抗菌药物和治疗方法提供有价值的理论依据。

Bacterial pneumonia caused by P. aeruginosa has become a global problem that threatens human health. Since the increasing rates of antibiotic resistance, it is urgent to find alternative treatment strategies. Currently, application of novel immunoregulatory factors serves as new clinical therapies for bacterial pneumonia. Previous studies from our group have elucidated PK2 exert beneficial activities in a P. aeruginosa induced murine pneumonia model, as well as promote the recruitment and phagocytosis of neutrophils in mice, although the underlying mechanisms involving the protective roles of PK2 remain unknown. Therefore, we hypothesized that PK2 prevented mice from P. aeruginosa induced pneumonia through mediating the release of cytokines and chemokines from airway epithelial cells, thereby regulating neutrophil function and immune responses of host. In our present study, knock out (KO) mice and primary cell culture system will be applied for in vivo and in vitro research, respectively. Moreover, real-time quantitative PCR, immunofluorescence, flow cytometry, together with gene chips, Luminex based multiple protein detection and CRISPR/Cas 9 system for genome editing techniques will be introduced to reveal the immune mechanisms of PK2 modulating neutrophil functions, which will provide a solid basis for the development of antibacterial drugs targeting PK2.