大黄鱼IL-35调节巨噬细胞极化及其机制研究

抗炎细胞因子白细胞介素-35（IL-35）在调节T、B和巨噬细胞功能及维持机体免疫稳态方面发挥重要作用。前期发现大黄鱼IL-35可调节巨噬细胞极化为M2型，但有关大黄鱼IL-35对巨噬细胞极化的调节机制仍不清楚。本项目拟在前期基础上通过研究IL-35对大黄鱼巨噬细胞吞噬、杀菌、活性氧与一氧化氮水平、细胞增殖及凋亡的影响，明确大黄鱼IL-35调节巨噬细胞M2型极化中的功能；采用免疫共沉淀结合受体敲降或抗体封阻受体实验，鉴定大黄鱼IL-35的巨噬细胞受体；利用重组IL-35蛋白刺激大黄鱼巨噬细胞，研究IL-35信号通路中关键信号分子转录水平、蛋白磷酸化水平和启动子活性水平的变化，及效应分子转录变化，以揭示IL-35调节巨噬细胞极化的信号通路。预期结果对于认识鱼类IL-35调节巨噬细胞极化及机制具有重要意义，也将为大黄鱼养殖病害的免疫防治提供新的思路。

Interleukin-35 (IL-35), an anti-inflammatory cytokine, plays important roles in regulating the functions of T cells, B cells and macrophages, and maintaining the balance of immune system. Our previous studies indicate that IL-35 can modulate macrophages to M2 type in large yellow croaker (Larimichthys crocea). However, the immunoregulatory mechanisms of IL-35 on macrophage polarization are still unclear. On the basis of previous studies, this project intends to detect the influences of IL-35 on phagocytosis, bactericidal activity, the levels of reactive oxygen species and nitric oxide, cell proliferation and apoptosis of macrophages to clarify the immunoregulatory functions of IL-35 on macrophages M2 type polarization. Besides, Co-immunoprecipitation assay and RNA interference or antibody blocking assay will be used to identify IL-35 receptors on macrophages. In addition, the changes of key signal molecular at the levels of transcription, protein, phosphorylation and promoter activity and effector molecular transcriptions will be tested, so as to clarify the signal pathways of IL-35 on macrophage polarization. The expected results are of great significance in understanding fish IL-35 on macrophage polarization and its regulatory mechanisms, and will also provide new ideas to prevent diseases during large yellow croaker cultivation.