cGAS enhances the HTLV-1 reverse transcription intermediate ssDNA90-induced innate immune re-sponses
10.3760/cma.j.issn.0254-5101.2018.06.006
- VernacularTitle:cGAS促进HTLV-1反转中间体ssDNA90诱导的固有免疫应答
- Author:
Zhixiang GUO
1
;
Di SONG
;
Yue LIU
;
Yuhan CUI
;
Lingling MA
;
Yuhe GUAN
;
Bo YANG
;
Jie WANG
Author Information
1. 453003 新乡医学院河南省免疫与靶向药物重点实验室;453003 新乡医学院医学检验学院科研创新班
- Keywords:
cGAS;
HTLV-1;
DNA sensor;
Reverse transcription intermediate;
Innate immune re-sponse
- From:
Chinese Journal of Microbiology and Immunology
2018;38(6):434-439
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the role of cyclic GMP-AMP synthase (cGAS),a cytosolic DNA sensor,in regulating innate immune responses induced by reverse transcription intermediate of human T cell leukemia virus type 1 (HTLV-1). Methods (1)ssDNA90,the reverse transcription intermediate of HTLV-1,was transfected into HeLa cells to observe changes in the expression pattern of cGAS in transfected-HeLa cells with immunoblot assay. (2) HeLa cells were firstly transfected with cGAS-encoding plasmid and then ssDNA90 24 hours later. Real-time PCR was used to measure the expression of interferon ( IFN)-β, IFN-gamma-inducible protein 10 ( IP-10 ), regulated on activation, normal T cell expressed and secreted (RANTES) and tumor necrosis factor (TNF)-α. Immunoblot assay was performed to measure phosphorylated interferon regulatory factor 3 (IRF3) and p65. (3)cGAS expression was silenced by siRNA in HeLa and phorbol-12-myristate-13-acetate (PMA)-treated THP1 (PMA-THP1) cells and then ssDNA90 was transfect-ed into these cells 24 hours later. Real-time PCR was used to measure the expression of IFN-β,IP-10,RAN-TES and TNF-α. Immunoblot assay was performed to measure phosphorylated IRF3 and p65. Results Ex-pression of cGAS was increased in HeLa cells after ssDNA90 transfection. Compared with control cells, cGAS-transfected HeLa cells showed increased expression of IFN-β, IP-10, RANTES and TNF-α and en-hanced phosphorylation of IRF3 and p65 after ssDNA90 transfection. Compared with control cells,both HeLa and PMA-THP1 cells with silenced expression of cGAS showed impaired production of IFN-β,IP-10,RAN-TES and TNF-α after ssDNA90 transfection. Moreover,ssDNA90-induced phosphorylation of IRF3 and p65 were decreased after cGAS gene-knockdown. Conclusion cGAS might promote HTLV-1 RTI ssDNA90-in-duced innate immune responses.
