Role of Rotavirus Enterotoxin NSP4 in the Inflammatory Response in Murine Macrophage RAW 264.7 Cells
Journal of Bacteriology and Virology
;
: 221-230, 2016.
Article
in English
| WPRIM
| ID: wpr-228230
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Plasmids
/
Rotavirus Infections
/
RNA
/
Glycoproteins
/
Dinoprostone
/
Blotting, Western
/
Cytokines
/
Chromatography, Affinity
/
Interleukin-6
/
Interleukin-10
Type of study:
Prognostic study
Limits:
Humans
Language:
English
Journal:
Journal of Bacteriology and Virology
Year:
2016
Type:
Article
Similar
MEDLINE
...
LILACS
LIS