Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China.
The First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510700, Guangdong Province, China.
World J Gastroenterol. 2018 May 28;24(20):2173-2180. doi: 10.3748/wjg.v24.i20.2173.
To assess the effects of hepatitis E virus (HEV) on the production of type I interferons (IFNs) and determine the underlying mechanisms.
We measured the production of interferon (IFN)-alpha and -beta (-α/β) in genotype 3 HEV-infected C3A cells at different time points (0, 8, 12, 24, 48, 72 and 120 h) by enzyme-linked immunosorbent assay (ELISA). The expression levels of IFN-stimulated gene (ISG)15 in HEV-infected C3A cells at different time points were tested by western blotting. The plasmid-expressing open reading frame 3 (ORF3) or control plasmids (green fluorescent protein-expressing) were transfected into C3A cells, and the levels of IFN-α/β and ISG15 were evaluated, respectively. Furthermore, the plasmid-expressing ISG15 or small interfering RNA-inhibiting ISG15 was transfected into infected C3A cells. Then, the production of IFN-α/β was also measured by ELISA.
We showed that genotype 3 HEV could enhance the production of IFN-α/β and induce elevation of ISG15 in C3A cells. HEV ORF3 protein could enhance the production of IFN-α/β and the expression of ISG15. Additionally, ISG15 silencing enhanced the production of IFN-α/β. Overexpression of ISG15 resulted in the reduction of IFN-α/β.
HEV may promote production of IFN-α/β and expression of ISG15 ORF3 in the early stages, and increased ISG15 subsequently inhibited the production of IFN-α/β.
评估戊型肝炎病毒（HEV）对 I 型干扰素（IFN）产生的影响，并确定其潜在机制。
我们通过酶联免疫吸附试验（ELISA）检测了不同时间点（0、8、12、24、48、72 和 120 h）感染基因型 3 HEV 的 C3A 细胞中 IFN-α和-β（-α/β）的产生。通过 Western blot 检测了不同时间点感染 HEV 的 C3A 细胞中干扰素刺激基因（ISG）15的表达水平。转染表达开放阅读框 3（ORF3）的质粒或对照质粒（表达绿色荧光蛋白），分别评估 IFN-α/β和 ISG15 的水平。此外，转染表达 ISG15 的质粒或抑制 ISG15 的小干扰 RNA 进入感染的 C3A 细胞。然后，通过 ELISA 测量 IFN-α/β的产生。
我们表明，基因型 3 HEV 可以增强 IFN-α/β的产生，并诱导 C3A 细胞中 ISG15 的升高。HEV ORF3 蛋白可以增强 IFN-α/β的产生和 ISG15 的表达。此外，ISG15 沉默增强了 IFN-α/β的产生。ISG15 的过表达导致 IFN-α/β的减少。
HEV 可能在早期促进 IFN-α/β和 ISG15 ORF3 的产生，随后增加的 ISG15 抑制 IFN-α/β的产生。
