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目的表达和纯化HCoV-229E的S1蛋白片段(S1 417-547),分析其诱导的免疫应答。方法将纯化蛋白免疫小鼠,ELISA检测小鼠血清特异性抗体以及血清IL-4和IFN-γ含量,流式细胞术检测小鼠脾脏T淋巴细胞亚群分布,观察其诱导的免疫应答。结果表达蛋白经金属螯合获得纯化,并诱导小鼠产生了高滴度的抗体。脾脏CD4+和CD8+比例均升高,CD4+/CD8+比值下降;免疫鼠血清IFN-γ和IL-4水平显著升高。结论成功构建了HCoV-229E S1蛋白的表达载体,并在BL21(DE3)中得到了高效表达,表达蛋白免疫小鼠后诱导了明显的细胞和体液免疫应答。
Objective To express and purify the S1 protein fragment of HCoV-229E (S1 417-547) and analyze its induced immune response. Methods The purified protein was used to immunize mice. Serum specific antibody, serum IL-4 and IFN-γ levels were detected by ELISA. The distribution of T lymphocyte subsets in splenic lymphocytes was detected by flow cytometry and the immune response was observed. Results The expressed protein was purified by metal chelation and the mice were induced to produce high titers of antibodies. The proportion of CD4 + and CD8 + in spleen increased, and the ratio of CD4 + / CD8 + decreased. The levels of IFN-γ and IL-4 in serum of immunized mice increased significantly. Conclusion The expression vector of HCoV-229E S1 protein was successfully constructed and highly expressed in BL21 (DE3). The expressed protein immunized mice induced obvious cellular and humoral immune responses.