Recombinant core proteins of Japanese encephalitis virus as activators of the innate immune response.
Nitric oxide (NO) has been shown to suppress Japanese encephalitis virus (JEV) RNA synthesis, viral protein accumulation, and virus release from infected cells. In this article, the potential viral structural proteins as the activators of NO product were studied at the molecular level. First, the genomic region encoding the JEV structural proteins was cloned into a prokaryotic expression vector pET for high-level expression. After purification, these JEV recombinant proteins were added to macrophages to examine the productions of NO and pro-inflammatory mediators. In this study, the recombinant core protein, but not envelope (E), could trigger NO and pro-inflammatory mediators (TNF-alpha, IL-6, and IL-12) productions on macrophages. And their effects were about 85-95% relative to LPS-stimulated macrophages in a dose-dependent manner. Meanwhile, the rCore-2D could up regulate promoters of IL-8 and TNF-alpha via EGFP expression in reporter plasmid (IL-8p-EGFP and TNF-alphap-EGFP)-transfected cells by flow cytometric analysis. These results suggest that JEV core protein could regulate pro-inflammatory mediators and NO production, and may play a crucial role in the innate immunity for the host to restrict the initial stage of JEV infection.
Chen
Shu-O
SO
Department of Veterinary, College of Veterinary Medicine, National Chung-Hsing University, Taichung, 402, Taiwan.
Fang
Shih-Hua
SH
Shih
Dien-Yun
DY
Chang
Tien-Jye
TJ
Liu
Jau-Jin
JJ
D000818:Animals; D019677:Artificial Gene Fusion; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis*; D004664:Encephalitis Virus, Japanese / Q000235:genetics / Q000276:immunology*; D005260:Female; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D017737:Macrophages, Peritoneal / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009569:Nitric Oxide / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology*; D014758:Viral Core Proteins / Q000235:genetics / Q000276:immunology*
D016207:Cytokines; D011994:Recombinant Proteins; D014758:Viral Core Proteins; D049452:Green Fluorescent Proteins; D009569:Nitric Oxide