Enterovirus D68 protease 2A affects anti-viral interferon type Ⅰ pathway
10.3760/cma.j.issn.0254-5101.2019.06.001
- VernacularTitle:肠道病毒-D68 2A蛋白对抗病毒IFN-Ⅰ通路影响的研究
- Author:
Huiwen ZHENG
1
;
Zhiyao YANG
;
Zening YANG
;
Jie SONG
;
Xing HUANG
;
Nan LI
;
Lisha DING
;
Heng LI
;
Hongzhe LI
;
Lei GUO
;
Manman CHU
;
Haijing SHI
;
Longding LIU
Author Information
1. 中国医学科学院&北京协和医学院医学生物学研究所
- Keywords:
EV-D68 infection;
Protease 2A;
Anti-viral IFN-Ⅰ pathway
- From:
Chinese Journal of Microbiology and Immunology
2019;39(6):401-409
- CountryChina
- Language:Chinese
-
Abstract:
Objective To analyze how enterovirus D68 (EV-D68) protease 2A affects the anti-vi-ral interferon typeⅠ(IFN-Ⅰ) pathway in 293T cells following infection. Methods Western blot was used to detect the expression of recombinant protease 2A, IFN-α and signal transducers and activators of tran-scription 1 (STAT1) at protein level. Expression of EV-D68 viral protein (VP1) and protease 2A was ana-lyzed by immunofluorescence at different time points. Cytopathic effects were recorded to calculate 50％ cell culture infective dose ( CCID50 ) . Expression of the genes involved in the anti-viral IFN-Ⅰ pathway was measured by real-time PCR (RT-PCR). Results The recombinant plasmid pCLIPf-2A was successfully constructed and the expression of recombinant protease 2A could be detected by Western blot 24 h after transfection. The recombinant protease 2A promoted the proliferation of EV-D68 at the late stage of infection and induced the production of IFN-α. Expression of the genes involved in the anti-viral IFN-Ⅰ pathway at mRNA level was up- or down-regulated to different degrees with various trends in different groups following infection. Expression of STAT1 was enhanced in all groups. Conclusions EV-D68 protease 2A promoted the activation of anti-viral IFN-Ⅰpathway in response to viral infection and enhanced the proliferation of virus at the late stage of infection.
