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目的利用显微成像技术拍摄并分析人偏肺病毒(human metapneumovirus,hMPV)的入胞过程,以探讨hMPV的感染免疫方式。方法使用全内反射荧光技术(total internal reflection fluorescence,TIRF)连续拍摄带有绿色荧光蛋白的病毒影像,并逐帧分析病毒的感染过程。结果感染开始后邻近培养介质的细胞膜上hMPV数量缓慢增多并在15～20 min时大量进入细胞。黏附于细胞上的病毒大部分做小振幅的无规律运动,偶见快速移动。感染12 h后hMPV在细胞膜阴影处、两细胞交界面大量聚集。结论本研究探讨了使用TIRF实时观察病毒入胞的可行性,结果表明TIRF技术能够连续观察病毒的运动规律和分布状况,为进一步揭示hMPV的感染免疫机制以及疫苗研制奠定了实验基础。
OBJECTIVE: To investigate the invasion and immunization of hMPV by microscopic imaging and to analyze the process of human metapneumovirus (hMPV) entry. Methods The total internal reflection fluorescence (TIRF) was used to continuously capture the green fluorescent protein-containing virus images and the virus infection was analyzed frame by frame. Results The number of hMPV on the cell membrane adjacent to the culture media slowly increased after infection started and entered the cells in large quantities at 15-20 min. Most of the viruses attached to cells do irregular movements of small amplitude, and occasionally move quickly. After 12 h infection hMPV in the cell membrane shadow Department, a large number of two cell interface aggregation. Conclusions This study explored the feasibility of using TIRF to observe the virus entering cells in real time. The results showed that the TIRF technique could continuously observe the movement and distribution of the virus and lay the experimental foundation for revealing the immune mechanism of hMPV infection and vaccine development.