呼吸道合胞病毒感染A549细胞后对Ⅰ型干扰素抗病毒免疫应答的影响  

Effect of respiratory syncytial virus infection on typeⅠ interferon response in A549 cells

作　　者：杨丽[1,2] 金鑫 余福勋[2] 张梅 余本莉 崔玉霞[4] 叶芝旭 YANG Li;JIN Xin;YU Fu-xun;ZHANG Mei;YU Ben-li;CUI Yu-xia;YE Zhi-xu(Department of Biomedicine,Guizhou Universily,Guiyang 5500001 Guizhou Province,China;不详)

机构地区：[1]贵州大学生物医学系,贵州贵阳550000 [2]中国卫生健康委员会肺脏免疫性疾病重点实验室,贵州贵阳550002 [3]贵州省人民医院眼科,贵州贵阳550002 [4]贵州省人民医院儿科,贵州贵阳550002

出　　处：《微生物学免疫学进展》2021年第6期8-12,共5页Progress In Microbiology and Immunology

基　　金：国家自然科学基金地区基金(81860003、81960001);贵州省科技厅支撑计划([2018]2785);贵州省中医药管理局中医药、民族医药科学技术研究课题(QZYY-2018-010);贵州省卫生计生委科学技术基金项目(gzwjw2018-1-049)。

摘　　要：目的研究呼吸道合胞病毒(respiratory syncytial virus,RSV)感染人肺腺癌细胞A549(简称A549)细胞后对I型干扰素抗病毒免疫应答的影响。方法RSV感染A549细胞,显微镜观察感染后细胞病变效应,免疫荧光检测RSV N蛋白表达以验证病毒感染成功。RT-PCR检测感染后12~48 h细胞中STAT1、IFN-α、IFN-βmRNA表达及ELISA检测细胞培养上清中IP-10、IFN-α、IFN-β、IFN-γ水平。结果RSV感染A549细胞后48 h形成明显的合胞病变,免疫荧光检测到大量RSV N蛋白表达。RSV感染后STAT1 mRNA及IP-10分泌水平明显升高(P<0.05),但IFN-α及IFN-βmRNA分泌水平较对照组差异无统计学意义(P>0.05)。结论RSV感染A549细胞后诱导了以IP-10分泌升高的抗病毒状态,但并未诱导IFN-α、IFN-β产生,提示I型干扰素的合成对RSV感染气道上皮细胞后局部抗病毒防御不是最重要的。Objective To study the effect of respiratory syncytial virus(RSV) infection on type I interferon antiviral immune response of A549 cells. Methods After infected by RSV, the cytopathic effect of A549 cells was observed by microscope, and the expression of RSV N protein was detected by immunofluorescence to verify the success of virus infection. RT-PCR was applied to determine the transcription levels of STAT1、IFN-α、IFN-β mRNA in cell culture during 12 to 48 hours post infection, and ELISA was applied to determine the levels of IP-10, IFN-α, IFN-β and IFN-γ in the supernatant. Results The cytopathic effect of RSV was obvious at 48 h post A549 infection, and a high level of RSV N protein was detected by immunofluorescence. The level of STAT1 mRNA and IP-10 increased significantly after RSV infection(P<0.05). However, both the mRNA expression and secretion of IFN-α and IFN-β were not significantly increased compared to the control group(P>0.05). Conclusion RSV infection of A549 cells induced an antiviral state by increasing the secretion of IP-10;however, it did not induce the secretion of IFN-α and IFN-β. This suggests that the synthesis of type I interferon is not the most important for the local antiviral defense post RSV infection of epithelial cells.

关 键 词：呼吸道合胞病毒 I型干扰素 干扰素-γ诱导蛋白10 抗病毒免疫应答 

分 类 号：R373.1[医药卫生—病原生物学]