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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.
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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

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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