Liaoning Key Laboratory of Marine Animal Immunology, Dalian Ocean University, Dalian, 116023, China; Liaoning Key Laboratory of Marine Animal Immunology and Disease Control, Dalian Ocean University, Dalian, 116023, China.
Liaoning Key Laboratory of Marine Animal Immunology, Dalian Ocean University, Dalian, 116023, China; Liaoning Key Laboratory of Marine Animal Immunology and Disease Control, Dalian Ocean University, Dalian, 116023, China; Dalian Key Laboratory of Disease Control and Prevention of Aquaculture Animals, Dalian Ocean University, Dalian, 116023, China.
Dev Comp Immunol. 2019 Jun;95:28-37. doi: 10.1016/j.dci.2019.01.013. Epub 2019 Jan 31.
The sp1A/ryanodine receptor (SPRY) family members have been reported to involve in important biological pathways, including innate immune signaling, cytokine signaling suppression, development, cell growth, and retroviral restriction. In the present study, a SPRY domain-containing SOCS box protein (named as CgSPSB3) was identified and characterized from oyster Crassostrea gigas. The open reading frame of CgSPSB3 gene was of 699 bp, encoding a polypeptide of 232 amino acid residues with a central SPRY domain and a C-terminal SOCS box motif. CgSPSB3 mRNA transcripts could be detected in all the examined tissues with the highest level in hemocytes, which was about 82.72-fold (p < 0.05) of that in gonad. Furthermore, the expression level of CgSPSB3 mRNA in granulocytes was significantly higher than that in semi-granulocytes and agranulocytes, which was about 2.04-fold (p < 0.05) of the average level of hemocytes. Immunofluorescence assay further revealed that CgSPSB3 protein was mainly distributed in the cytoplasm of granulocytes. The mRNA expression of CgSPSB3 in hemocytes was up-regulated after lipopolysaccharide (LPS) and Vibrio splendidus stimulations. The mRNA expression of CgIFNLP, CgIL17-5 and CgTNF-1 decreased significantly (p < 0.05) at 24 h after the CgSPSB3 mRNA was knocked down by RNAi. These results collectively indicated that CgSPSB3 might play an important role in regulating cytokines production in granulocytes of C. gigas.
SPRY 家族成员已被报道参与重要的生物学途径，包括先天免疫信号、细胞因子信号抑制、发育、细胞生长和逆转录病毒限制。本研究从牡蛎(Crassostrea gigas)中鉴定并表征了一个含有 SPRY 结构域的 SOCS 盒蛋白(CgSPSB3)。CgSPSB3 基因的开放阅读框为 699bp，编码一个 232 个氨基酸残基的多肽，其中央含有 SPRY 结构域，C 端含有 SOCS 盒基序。CgSPSB3 mRNA 转录本可在所有检测到的组织中检测到，在血细胞中的表达水平最高，约为性腺的 82.72 倍(p<0.05)。此外，粒细胞中 CgSPSB3 mRNA 的表达水平明显高于半粒细胞和无粒细胞，约为血细胞平均水平的 2.04 倍(p<0.05)。免疫荧光分析进一步表明，CgSPSB3 蛋白主要分布在粒细胞的细胞质中。LPS 和灿烂弧菌刺激后，血细胞中 CgSPSB3 的 mRNA 表达上调。用 RNAi 敲低 CgSPSB3 mRNA 后，24h 时 CgIFNLP、CgIL17-5 和 CgTNF-1 的 mRNA 表达显著降低(p<0.05)。这些结果共同表明，CgSPSB3 可能在调节牡蛎血细胞中细胞因子的产生中发挥重要作用。
