title: Immune responses in the Japanese pufferfish (Takifugu rubripes) head kidney cells stimulated with particulate silica.
abstract: Studies on immune response to crystal silica in mammals indicate immune stimulation effect of environmental parameters including silica or asbestos, but there is no information on this aspect in lower vertebrates. Therefore, we examined expression of cytokine genes related to innate immunity in the Japanese pufferfish, Fugu (Takifugu rubripes) head kidney (HK) cells stimulated with particulate silica at 10 and 50 μg mL(-1). Expression of eleven cytokine genes was analyzed by the multiplex RT-PCR method (GenomeLab Genetic Analysis System, GeXPS; Beckman Coulter Inc.). Additionally, to confirm functionality of activated inflammatory immunity, we assessed phagocytic activity. Expression of NLR family genes as potential sensor molecules of inflammasome and inflammasome-associated genes (ASC and caspase-1) was also confirmed in HK cells by quantitative real-time PCR (qRT-PCR). As a result, an increased gene expression of pro-inflammatory cytokines (IL-6, IL-17A/F3, TNF-α, TNF-β and IFN-γ) and other cytokines (IL-4/13A, IL-4/13B, Type I-IFN) was recorded in particulate silica stimulated HK cells. Moreover, phagocytic activity showed a tendency to significantly increase in stimulated monocyte of HK cells after 6 h. Expression of NLR-C9 and NLR-C12 genes significantly increased in silica-stimulated HK cells. The particulate silica also significantly induced expression of inflammosome-associated genes, which may relate to the induced NLR-Cs.
authors.affiliation: Department of Biochemistry and Applied Biosciences, Faculty of Agriculture, University of Miyazaki, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Morimoto
authors.affiliation: Interdisciplinary Graduate School of Agriculture and Engineering, University of Miyazaki, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan.
authors.forename: Gouranga
authors.initials: G
authors.lastname: Biswas
authors.affiliation: Department of Biochemistry and Applied Biosciences, Faculty of Agriculture, University of Miyazaki, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan.
authors.forename: Tomoya
authors.initials: T
authors.lastname: Kono
authors.affiliation: Department of Biochemistry and Applied Biosciences, Faculty of Agriculture, University of Miyazaki, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Sakai
authors.affiliation: Department of Biochemistry and Applied Biosciences, Faculty of Agriculture, University of Miyazaki, 1-1 Gakuenkibanadai-nishi, Miyazaki 889-2192, Japan. Electronic address: jhikima@cc.miyazaki-u.ac.jp.
authors.forename: Jun-Ichi
authors.initials: J
authors.lastname: Hikima
mesh_terms: D000818:Animals; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D005393:Fish Diseases / Q000139:chemically induced / Q000276:immunology*; D029941:Fish Proteins / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D060226:Head Kidney / Q000276:immunology / Q000378:metabolism; D058847:Inflammasomes; D007249:Inflammation / Q000139:chemically induced / Q000276:immunology / Q000662:veterinary*; D010587:Phagocytosis; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D012822:Silicon Dioxide / Q000494:pharmacology*; D023741:Takifugu / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D016207:Cytokines; D029941:Fish Proteins; D058847:Inflammasomes; D012822:Silicon Dioxide
keywords: ASC; Caspase-1; Inflammatory cytokines; Japanese pufferfish; Multiplex RT-PCR; Particulate silica