TBEV对人神经细胞致病性的实验研究

Studying the pathogenicity of TBEV to human neuroblastoma tumor cells

目的确定蜱传脑炎病毒（Tick-borne encephalitis virus，TBEV）对人神经母瘤细胞的感染性及复制增殖情况。方法将TBEV感染人神经母瘤细胞SK－N－SH，取不同时间点的细胞培养上清，分别采用real－timeRT－PCR方法及空斑测定的方法测定病毒滴度，以确定TBEV在SK－N－SH细胞中的复制增殖情况；同时进行细胞形态观察及免疫荧光检测，以确定TBEV感染后的SK－N－SH细胞形态变化。结果TBEV感染SK－N－SH细胞后，可进行大量复制增殖，感染后3d，病毒滴度达到峰值，可达2．92×10＾7PFU／ml，感染细胞产生明显的形态改变，免疫荧光法可检测到细胞内的病毒粒子。结论TBEV可在人神经母瘤细胞SK－N－SH中大量复制增殖，并可造成细胞明显的形态改变，因此人神经母瘤细胞SK-N-SH可以作为TBEV培养良好的细胞模型。

Objective To identify the infection and the replication of Tick-borne encephalitis virus （TBEV） in human neuroblastoma cells. Methods After being infected with TBEV, the cell culture supernatant of human neuroblastoma cell line SK-N-SH was collected and assayed at different time points. By using real-time RT-PCR and plaque assay to measure the titer of virus in the supernatant, the replication and proliferation of TBEV in human neuroblastoma cell was identified. Meanwhile, the morphological change of SK-N-SH after TBEV infection was also visualized by observation under microscope and immunofluoreseence assay. Results Real-time RT-PCR and plaque assay both demonstrated that TBEV could replicate effectively in SK-N-SH cells, the peak titer could reach 2.92×107 PFU/ml on 3 days post-inoculation. And significant morphological change occured on infected SK-N-SH cells after 2 days post inoculation. By immunofluoreseence assay, the virus particles could be detected and visualized. Conclusion TBEV can replicate and proliferate effectively and cause significant cell morphological changes in human neuroblastoma cell SK-N- SH, which demonstrated that SK-N-SH could be a suitable cell model for TBEV culture.