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Partial sequences of cytokine genes IFNγ, IL-1β, IL-8, IL-10, and TNFα were identified from next-generation sequencing data for three amberjack species: Seriola quinqueradiata, S. lalandi and S. dumerili. The cytokine genes had more than 90 % nucleotide identity among the three species. Quantitative PCR (qPCR) primers were designed from the consensus sequences to detect cytokine genes. The specificities of consensus qPCR primers were investigated in the head kidney of three amberjack species treated with formalin-killed cells (FKC) of Vibrio anguillarum and polyinosinic-polycytidylic acid [poly(I:C)] at 3 h post injection (hpi) using qPCR. The primers detected up-regulation of certain cytokine genes after FKC of V. anguillarum and poly(I:C) stimulation in all species examined. Consequently, the primers are suitable for studying and detecting cytokine genes after vaccination and pathogen infection in these three amberjack species. [ABSTRACT FROM AUTHOR]

Abstract: The mucosal surfaces of fish allow for the introduction of foreign substances, including antigens, from the surrounding environment. In this study, uptake of Vibrio anguillarum J-O-3 serotype bacterin by Japanese flounder, and the subsequent immune responses were investigated. Immunohistochemistry revealed that the bacterin was taken up through the epithelial cells of gills. The transcription levels of inflammatory cytokines such as interleukin (IL)-1β, IL-6 and tumor necrosis factor α were significantly up-regulated in the gills at 3 days following exposure to the bacterin. There was also a corresponding increase in IL-8 receptor, CD4-1, CD4-2 and CD8α transcript levels in the gills. Our findings suggest that the gills play a major role in the uptake of V. anguillarum bacterin and induction of inflammation, which results in an activation of the adaptive immune response in teleost fish. [Copyright &y& Elsevier]