Role of Rotavirus Enterotoxin NSP4 in the Inflammatory Response in Murine Macrophage RAW 264.7 Cells.
10.4167/jbv.2016.46.4.221
- Author:
Song Ah KIM
1
;
Van Thai THAN
;
Wonyong KIM
Author Information
1. Department of Microbiology, Chung-Ang University College of Medicine, Seoul, Korea. kimwy@cau.ac.kr
- Publication Type:Original Article
- Keywords:
Rotavirus;
NSP4;
RAW 264.7 murine macrophage;
Inflammation
- MeSH:
Blotting, Western;
Chromatography, Affinity;
Clone Cells;
Cytokines;
Diarrhea;
Dinoprostone;
DNA, Complementary;
Electrophoresis, Polyacrylamide Gel;
Enterotoxins*;
Glycoproteins;
Humans;
Inflammation;
Interleukin-10;
Interleukin-6;
Macrophages*;
Nitric Oxide;
Plasmids;
RAW 264.7 Cells*;
RNA;
Rotavirus Infections;
Rotavirus*
- From:Journal of Bacteriology and Virology
2016;46(4):221-230
- CountryRepublic of Korea
- Language:English
-
Abstract:
The rotavirus nonstructural glycoprotein, NSP4, has been identified as the first viral enterotoxin capable of inducing diarrhea. To investigate the biological function of NSP4 in the inflammatory process, a cDNA from human rotavirus (Wa strain) RNA segment 10 was amplified by RT-PCR, cloned into TA vector, and subsequently subcloned into pET23b expression plasmid. The expression of NSP4 protein was determined by SDS-PAGE and Western blotting, then, the protein was purified by affinity chromatography on Ni-NTA-agarose column. The inflammatory effects of NSP4, namely, production of nitric oxide (NO), pro-inflammatory cytokines (IL-1β, IL-6, IL-10, and TNF-α), and prostaglandin E2 (PGE₂), was evaluated using NSP4-stimulated RAW 264.7 murine macrophages and compared with those observed after stimulation with lipopolysaccharide (LPS). The levels of IL-1β, IL-6, and TNF-α were significantly increased, and those of NO and PGE₂ also increased in NSP4-stimulated RAW 264.7 cells. These findings indicate that NSP4 plays an important role in the inflammatory response observed during rotavirus infection.
