Characterization and expression analysis of rockfish (Sebastes schlegelii) myeloid differentiation factor-88 (SsMyD88) and evaluation of its ability to induce inflammatory cytokines through NF-ĸB
| Autor: | M.D. Neranjan Tharuka, K.A.S.N. Shanaka, Hyerim Yang, Jehee Lee, Thanthrige Thiunuwan Priyathilaka, Sarithaa Sellaththurai |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Fish Proteins
Lipopolysaccharides 0301 basic medicine Gene Expression Aquatic Science Biology Proinflammatory cytokine Mice 03 medical and health sciences Immune system Antigen Animals Humans Environmental Chemistry Receptor Transcription factor Inflammation Innate immune system Macrophages NF-kappa B Interleukin 04 agricultural and veterinary sciences General Medicine Immunity Innate Perciformes Cell biology HEK293 Cells Poly I-C RAW 264.7 Cells 030104 developmental biology Myeloid Differentiation Factor 88 040102 fisheries Cytokines 0401 agriculture forestry and fisheries Streptococcus iniae |
| Zdroj: | Fish & Shellfish Immunology. 99:59-72 |
| ISSN: | 1050-4648 |
| DOI: | 10.1016/j.fsi.2020.01.060 |
| Popis: | Innate immunity is characterized by nonspecific, prompt reactions toward armada of antigens. Animals funnel down a repertoire of immune stimulants to activate non-selective defense mechanisms rapidly. This study was conducted to characterize the rockfish (Sebastes schlegelii) adaptor protein MyD88 (SsMyD88), which interacts with both toll-like receptors and interleukin receptors. The tissue expression of unchallenged SsMyD88 was evaluated by quantitative real time PCR (qPCR). Fish were intraperitoneally injected with immune stimulants including poly I:C, lipopolysaccharides, and Streptococcus iniae. Then, the temporal expression of SsMyD88 was analyzed. Finally, the inflammatory gene expression and downstream promoter activation were analyzed. Strongest expressions were reported in the liver, gills and spleen in unchallenged conditions. All diverse immune stimulants were found to be capable of significantly altering SsMyD88 transcription during the challenge experiment. Evaluation of downstream promoter biases by SsMyD88 found a predominant activation of NF-ĸB transcription factors when compared with the AP-1, revealing significant and substantial upregulation of major inflammatory mediators such as IL-1-β, IL-6, iNOS, COX-2 and TNF-α. Fluorescent detection confirmed an intense production of NO and the predominant differentiation of macrophages into M1 lineage with the overexpression of SsMyD88 in vitro. These results further corroborate the role of SsMyD88 as a mediatory molecule that bridges distinct immune stimulants to induce drastic immune responses in fish. |
| Databáze: | OpenAIRE |
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