摘要
目的探讨DNA识别受体干扰素诱导蛋白16(IFI16)在成人T淋巴细胞白血病1型病毒(HTLV-1)阳性T细胞中对HTLV-1病毒蛋白和促炎因子表达的影响。方法在不同的HTLV-1阳性T细胞系中,采用免疫印迹试验检测IFI16在HTLV-1阳性T细胞中的表达变化;构建IFI16基因沉默siRNA并用免疫印迹检测其IFI16敲低表达的效率;用siRNA在HTLV-1阳性T细胞系中敲低IFI16表达后,通过real-time PCR和采用免疫印迹分别检测HTLV-1病毒蛋白Tax在mRNA和蛋白质水平上的表达量;并通过real-time PCR检测干扰素(IFN)-α、IFN-γ、肿瘤坏死因子(TNF)-α和HTLV-1病毒基因Tax以及Env的表达量。结果与HTLV-1病毒阴性T细胞Jurkat相比,IFI16在HTLV-1阳性T细胞MT2、MT4以及C8166中表达均增高;在MT2和MT4细胞中用siRNA敲低IFI16表达后,HTLV-1病毒蛋白Tax表达量上升,IFN-α、IFN-γ以及TNF-α的表达量下降。结论在HTLV-1病毒稳定感染的MT2和MT4细胞中,IFI16表达量上升,从而促进干扰素和促炎因子的产生,进而抑制HTLV-1病毒蛋白的表达。
Objective To investigate the effects of interferon inducible protein 16 (IFI16), a cytosolic DNA sensor, on the expression of human T-cell leukemia virus type 1 (HTLV-1) proteins and pro-inflammatory cytokines in adult HTLV-1-positive T cells. Methods IFI16 expression in different HTLV-1-positive T cell lines was detected by immunoblot assay. Specific siRNA targeting the IFI16 gene was constructed and the gene silencing efficiency was detected by immunoblot assay. Expression of HTLV-1 Tax protein at mRNA and protein levels was respectively detected by real-time PCR and immunoblot assay after knocking down the expression of IFI16 in HTLV-1-positive T cells with siRNA. Expression of interferon (IFN)-α, IFN-γ, tumor necrosis factor (TNF)-α, Tax and Env were detected by real-time PCR. Results Compared with the HTLV-1-negative T cell line Jurkat, IFI16 expression was enhanced in the HTLV-1-positive T cell lines MT2, MT4 and C8166. Tax expression was increased, while that of IFN-α, IFN-γ and TNF-α was decreased in MT2 and MT4 cells after silencing the expression of IFI16 with siRNA. Conclusions IFI16 expression was increased in HTLV-1-positive MT2 and MT4 cells. Meanwhile, IFI16 promoted the production of interferon and pro-inflammatory cytokines and inhibited the expression of HTLV-1 proteins.
作者
崔钰晗
关宇鹤
刘月
张葛
陈凡
陈蒙蒙
孙静靓
任晓文
杨波
王洁
Cui Yuhan;Guan Yuhe;Liu Yue;Zhang Ge;Chen Fan;Chen Mengmeng;Sun Jingliang;Ren Xiaowen;Yang Bo;Wang Jie(Henan Key Laboratory of Immunology and Targeted Drugs, Xinxiang Medical University, Xinxiang 453003, China;Scientific Research Innovation Group, School of Laboratory Medicine, Xinxiang Medical University, Xinxiang 453003, China;Collaborative Innovation Center of Molecular Diagnosis and Laboratory Medicine in Henan Province, Xinxiang 453003, China)
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2019年第9期668-673,共6页
Chinese Journal of Microbiology and Immunology
