论文部分内容阅读
目的研究H5N1病毒HA蛋白作用于小鼠后对其外周免疫器官脾脏的损伤其相关细胞因子的分泌。方法通过昆虫-杆状病毒表达系统表达H5N1病毒HA蛋白,并经镍亲和磁珠纯化,将重组的HA蛋白作用于小鼠,72 h后,取其脾脏,观察其镜下形态学变化;分离脾细胞并培养,检测其细胞因子的分泌。结果脾组织病理表现为局部性弥漫性肺泡损伤,单核细胞侵润,脾脏肿胀,毛细血管充血,生发中心局部结构破坏,有空泡样改变,淋巴细胞减少,见较多大单核吞噬细胞。HA蛋白预处理的小鼠其脾细胞分泌IP-10、MCP-1a、Rantes较PBS预处理的小鼠明显增高;同时HA蛋白预处理组小鼠脾细胞在LPS刺激后分泌的IFN-γ、IP-10、MCP-1a、Rantes较PBS处理的小鼠其脾细胞明显增高。结论气道给予H5N1 HA表面膜蛋白能够始动异常的免疫应答,引起急性肺损伤和外周淋巴器官与免疫细胞破坏。
Objective To study the secretion of related cytokines in the spleen of peripheral immune organs after H5N1 virus HA protein acts on mice. Methods The HA protein of H5N1 virus was expressed by the insect - baculovirus expression system and purified by nickel affinity magnetic beads. The recombinant HA protein was applied to mice for 72 h. The spleens were harvested and the morphological changes were observed. Spleen cells were isolated and cultured to detect the secretion of cytokines. Results The pathological changes of splenic tissue were local diffuse alveolar injury, mononuclear cell infiltration, spleen swelling, capillary congestion, local structure destruction in germinal centers, vacuolar changes and lymphopenia, and more mononuclear phagocytes. HA protein pretreatment mice spleen cells secreted IP-10, MCP-1a, Rantes significantly increased compared with PBS pretreatment mice; at the same time HA protein pretreatment group splenocytes secreted IFN-γ after LPS stimulation, The spleen cells of IP-10, MCP-1a and Rantes were significantly higher than those of PBS-treated mice. Conclusion Airway administration of H5N1 HA surface membrane protein initiates an abnormal immune response, causing acute lung injury and destruction of peripheral lymphoid organs and immune cells.