一种干扰素基因（CgSTING）刺激物，参与牡蛎 Crassostrea gigas 的抗菌免疫反应。
A stimulator of interferon gene (CgSTING) involved in antimicrobial immune response of oyster Crassostrea gigas.
机构信息
Liaoning Key Laboratory of Marine Animal Immunology, Dalian Ocean University, Dalian, 116023, China; Functional Laboratory of Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266235, 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; Southern Laboratory of Ocean Science and Engineering (Guangdong, Zhuhai), Zhuhai, 519000, China; Functional Laboratory of Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266235, China; Liaoning Key Laboratory of Marine Animal Immunology and Disease Control, Dalian Ocean University, Dalian, 116023, China; Dalian Key Laboratory of Aquatic Animal Disease Prevention and Control, Dalian Ocean University, Dalian, 116023, China.
出版信息
Fish Shellfish Immunol. 2022 Sep;128:82-90. doi: 10.1016/j.fsi.2022.07.059. Epub 2022 Jul 30.
The stimulator of interferon gene (STING), an intracellular sensor of cyclic dinucleotides, is critical to the innate immune response, especially the induction of type I interferon (IFN) during pathogenic infection. A STING homologue (CgSTING) regulating the expression of IFN-like protein (CgIFNLP) was previously identified in the Pacific oyster Crassostrea gigas, and its involvement in antibacterial immunity was further investigated in the present study. The mRNA transcripts of CgSTING were ubiquitously detected in all the three subpopulations of haemocytes with the highest expression in semi-granulocytes. After the stimulation with Vibrio splendidus, the mRNA expression of CgSTING in haemocytes was significantly up-regulated and peaked at 72 h, which was 12.91-fold of that in control group (p < 0.01). The CgSTING protein was mainly located in the cytoplasm of haemocytes. After the expression of CgSTING was knocked down (0.12-fold of that in control group, p < 0.05) by RNAi, the mRNA expression levels of interleukin17-1 (CgIL17-1), interleukin17-3 (CgIL17-3), interleukin17-4 (CgIL17-4), defensins (Cgdefh1, Cgdefh2), big defensin (CgBigDef1), interferon-like protein (CgIFNLP), tumor necrosis factor (CgTNF) and nuclear factor-κB (CgRel) all decreased significantly at 12 h after V. splendidus stimulation, which was 0.12-fold-0.72-fold (p < 0.05) of that in control group, respectively. The positive signals of CgRel were observed in the haemocyte nucleus after V. splendidus stimulation. The nuclear translocation of CgRel was suppressed in CgSTING-RNAi oysters, and the green signals of CgRel were mainly observed in the haemocyte cytoplasm after V. splendidus stimulation. Furthermore, the number of V. splendidus in the haemolymph of CgSTING-RNAi oysters increased significantly, which was 26.78-fold (p < 0.01) of that in the control group at 12 h after V. splendidus stimulation. These results indicated that CgSTING played important role in the immune defense against bacterial infection by inducing the expressions of cytokines and defensins.
干扰素基因刺激物（STING）是一种细胞内环二核苷酸的感应器，是先天免疫反应的关键，尤其是在致病性感染期间诱导 I 型干扰素（IFN）的产生。先前在太平洋牡蛎（Crassostrea gigas）中鉴定出一种调节 IFN 样蛋白（CgIFNLP）表达的 STING 同源物（CgSTING），本研究进一步研究了其在抗菌免疫中的作用。CgSTING 的 mRNA 转录本在三种血淋巴细胞亚群中广泛存在，其中半颗粒细胞中的表达最高。在受到灿烂弧菌刺激后，血细胞中 CgSTING 的 mRNA 表达水平显著上调，在 72 小时达到峰值，是对照组的 12.91 倍（p<0.01）。CgSTING 蛋白主要位于血细胞的细胞质中。用 RNAi 敲低 CgSTING 的表达（对照组的 0.12 倍，p<0.05）后，在灿烂弧菌刺激后 12 小时，白细胞介素 17-1（CgIL17-1）、白细胞介素 17-3（CgIL17-3）、白细胞介素 17-4（CgIL17-4）、防御素（Cgdefh1、Cgdefh2）、大防御素（CgBigDef1）、干扰素样蛋白（CgIFNLP）、肿瘤坏死因子（CgTNF）和核因子-κB（CgRel）的 mRNA 表达水平均显著降低，分别为对照组的 0.12 倍-0.72 倍（p<0.05）。在灿烂弧菌刺激后，可观察到 CgRel 的阳性信号位于血细胞核内。在 CgSTING-RNAi 牡蛎中，CgRel 的核转位受到抑制，在灿烂弧菌刺激后，CgRel 的绿色信号主要观察到在血细胞细胞质中。此外，在灿烂弧菌刺激后 12 小时，CgSTING-RNAi 牡蛎的血淋巴中灿烂弧菌的数量显著增加，是对照组的 26.78 倍（p<0.01）。这些结果表明，CgSTING 通过诱导细胞因子和防御素的表达，在抵抗细菌感染的免疫防御中发挥重要作用。
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