title: TNF-α is involved in apoptosis triggered by grass carp reovirus infection in vitro.
abstract: Grass carp reovirus (GCRV) infection causes apoptosis in Ctenopharyngodon idella kidney cells (CIK). However, the cause of GCRV-induced apoptosis and its signaling pathways remain unknown. This study investigated the role of TNF-α-induced capase-8 pathways in mediating GCRV-induced apoptosis in the grass carp (Ctenopharyngodon idella). Recombinant TNF-α was expressed and purified from Escherichia. coli. The western blot assay indicated that TNF-α expression level in kidney and spleen was higher than that in liver. In apoptosis assay, recombinant TNF-α triggered significant apoptosis in CIK cells, which was characterized by increased mRNA levels of TNF-α, TRADD or caspase-8, and enhanced caspase-8 activity in CIK cells. To confirm the biological activity of TNF-α during GCRV infection, significant apoptosis in CIK cells was induced by GCRV correlating with enhanced caspase-8 activity, increased mRNA level of TNF-α, TRADD or caspase-8, increased protein level of TNF-α in CIK cells and cell supernatant, suggesting that TNF-α-induced capase-8 pathways might be involved in GCRV-triggered apoptosis. Furthermore, treatment with an anti-TNF-α polyclonal antibody significantly decreased the degree of apoptosis in infected CIK cells compared with cells treated with a control antibody, which confirmed that TNF-α was a key mediator involved in GCRV-induced apoptosis. Taken together, these results indicated that GCRV might trigger apoptosis via TNF-α induced capase-8 pathways in CIK cells.
authors.affiliation: Aquatic Pathogen Collection Center, MOA Key Laboratory of Freshwater Fishery Germplasm Resources, Shanghai Ocean University, Shanghai 201306, China.
authors.forename: Jianfei
authors.initials: J
authors.lastname: Lu
authors.affiliation: Aquatic Pathogen Collection Center, MOA Key Laboratory of Freshwater Fishery Germplasm Resources, Shanghai Ocean University, Shanghai 201306, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Aquatic Pathogen Collection Center, MOA Key Laboratory of Freshwater Fishery Germplasm Resources, Shanghai Ocean University, Shanghai 201306, China.
authors.forename: Zhaoyuan
authors.initials: Z
authors.lastname: Shen
authors.affiliation: Aquatic Pathogen Collection Center, MOA Key Laboratory of Freshwater Fishery Germplasm Resources, Shanghai Ocean University, Shanghai 201306, China.
authors.forename: Cuiyu
authors.initials: C
authors.lastname: Lu
authors.affiliation: Aquatic Pathogen Collection Center, MOA Key Laboratory of Freshwater Fishery Germplasm Resources, Shanghai Ocean University, Shanghai 201306, China. Electronic address: lqlv@shou.edu.cn.
authors.forename: Liqun
authors.initials: L
authors.lastname: Lu
mesh_terms: D000818:Animals; D017209:Apoptosis; D002347:Carps; D004926:Escherichia coli / Q000235:genetics; D005393:Fish Diseases / Q000276:immunology* / Q000821:virology; D029941:Fish Proteins / Q000235:genetics* / Q000378:metabolism; D007113:Immunity, Innate; D007668:Kidney / Q000276:immunology / Q000502:physiology; D009928:Organ Specificity; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012087:Reoviridae / Q000502:physiology; D012088:Reoviridae Infections / Q000276:immunology / Q000662:veterinary* / Q000821:virology; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics* / Q000378:metabolism
chemical_list: D029941:Fish Proteins; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha
keywords: Apoptosis; Capase-8; Grass carp reovirus; TNF-α; TRADD