【摘要】 目的:研究严重急性呼吸综合征(SARS)冠状病毒(CoV)S蛋白作用于呼吸道上皮及免疫细胞后对趋化因子IP-10和其他细胞因子生成的影响。方法:通过昆虫-杆状病毒表达系统表达SARS-CoV的S蛋白,并经镍亲和磁珠纯化,将重组的S蛋白分别作用于人支气管上皮细胞16HBE、外周血单个核细胞和单核细胞、肺泡单核-巨噬细胞,观察其形态学变化,并检测IP-10和其他细胞因子的变化。结果:基础状态下16HBE并不生成IP-10,S蛋白作用于人支气管上皮细胞16HBE,引起部分细胞变圆、空泡形成并裂解,12h后即可见IP-10生成达(79.97±13.81)pg/ml,呈显著量效关系。但对其他参与免疫反应的Th1/Th2细胞因子及趋化因子影响不明显。S蛋白并不诱导外周血单核细胞和单个核细胞(主要为淋巴细胞)合成释放IP-10。比较而言,干扰素-γ(IFN-γ)不仅能诱导16HBE,而能诱导免疫细胞生成IP-10。结论:①SARS-CoV可通过其S蛋白早期诱导肺上皮细胞合成释放IP-10,从而启动免疫病理过程。②SARS-CoVS蛋白诱导IP-10在宿主细胞的生成不依赖IFN-γ。更多还原

【Abstract】 Objective: To investigate the role of the recombinant S protein of SARS-CoV in the induction of chemokine IP-10 and other cytokines in airway epithelial cells and immunocytes. Methods: Using insect-baculovirus expression system and Nickel affinity Magnet Beads, S protein of SARS-CoV was produced and then used to stimulate cultured human bronchial epithelial cells (16HBE), human peripheral blood mononuclear cells(PBMC), human peripheral blood monocytes and alveolar macrophages. The levels of IP-10 and the cytokines involved in immunoreaction in response to virus infection were detected in the supernatants of those cells cultured with the S protein by liquid chip system. Results: Under normal condition, no detectable IP-10 was found in 16HBE. A high level of IP-10(79.97± 13.81) pg/ml was detected in the 16HBE 12 hrs after being treated with the S protein, and the induction of IP-10 by S protein displayed at a significant quantity-effect reaction, but not in PBMC, monocytes and alveolar macrophages. In contrast, IFN-γ was able to induce the production of IP-10 in either 16HBE or the immunocytes. Conclusion: 1.S protein of SARS-CoV can induce a high level of IP-10 in lung epithelial cells at early stage after the virus infection, which may initiate the process of the immune damage in the lung. 2. S protein of SARS-CoV induces the production of IP-10 by a way of IFN-γ independent.更多还原