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半滑舌鳎干扰素诱导蛋白 44 样(CgIFI44L)参与抗病毒免疫反应。

The involvement of an interferon-induced protein 44-like (CgIFI44L) in the antiviral immune response of Crassostrea gigas.

机构信息

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 Aquatic Animal Disease Prevention and 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; 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 Oct;129:96-105. doi: 10.1016/j.fsi.2022.08.064. Epub 2022 Aug 30.

Abstract

Interferon-stimulated genes (ISGs) encoding proteins are the essential executors of interferon (IFN) mediated antiviral defense. In the present study, an ISG member, interferon-induced protein 44-like (IFI44L) gene (designed as CgIFI44L-1) was identified from the Pacific oyster Crassostrea gigas. The ORF of CgIFI44L-1 cDNA was of 1437 bp encoding a polypeptide of 479 amino acids with a TLDc domain and an MMR_HSR1 domain. The mRNA transcripts of CgIFI44L-1 were detected in all the tested tissues with highest level in haemocytes, which was 15.78-fold of that in gonad (p < 0.001). Among the haemocytes, the CgIFI44L-1 protein was detected to be highly expressed in granulocytes with dominant distribution in cytoplasm. The mRNA expression level of CgIFI44L-1 in haemocytes was significantly induced by poly (I:C) stimulation, and the expression level peaked at 24 h, which was 24.24-fold (p < 0.0001) of that in control group. After the treatment with the recombinant protein of an oyster IFN-like protein (rCgIFNLP), the mRNA expression level of CgIFI44L-1 was significantly enhanced at 6 h, 12 h and 24 h, which was 2.67-fold (p < 0.001), 5.44-fold (p < 0.001) and 5.16-fold (p < 0.001) of that in control group, respectively. When the expressions of CgSTAT and CgIFNLP were knocked down by RNA interference (RNAi), the mRNA transcripts of CgIFI44L-1 were significantly down-regulated after poly (I:C) stimulation, which was 0.09-fold (p < 0.001) and 0.06-fold (p < 0.001) of those in EGFP group, respectively. These results suggested that CgIFI44L-1 was a conserved ISG in oyster, which was regulated by CgIFNLP and CgSTAT, and involved in the oyster antiviral immune response.

摘要

干扰素刺激基因(ISGs)编码的蛋白是干扰素(IFN)介导的抗病毒防御的必要执行者。在本研究中,从太平洋牡蛎(Crassostrea gigas)中鉴定出一个 ISG 成员,干扰素诱导蛋白 44 样(IFI44L)基因(命名为 CgIFI44L-1)。CgIFI44L-1 cDNA 的 ORF 为 1437 bp,编码 479 个氨基酸的多肽,具有 TLDc 结构域和 MMR_HSR1 结构域。CgIFI44L-1 的 mRNA 转录本在所有检测的组织中均有表达,在血细胞中表达水平最高,比性腺高 15.78 倍(p < 0.001)。在血细胞中,CgIFI44L-1 蛋白在粒细胞中高度表达,主要分布在细胞质中。聚(I:C)刺激后,血细胞中 CgIFI44L-1 的 mRNA 表达水平显著诱导,在 24 小时时达到峰值,是对照组的 24.24 倍(p < 0.0001)。在用牡蛎 IFN 样蛋白(rCgIFNLP)的重组蛋白处理后,CgIFI44L-1 的 mRNA 表达水平在 6、12 和 24 小时时显著增强,分别是对照组的 2.67 倍(p < 0.001)、5.44 倍(p < 0.001)和 5.16 倍(p < 0.001)。当通过 RNA 干扰(RNAi)敲低 CgSTAT 和 CgIFNLP 的表达时,聚(I:C)刺激后 CgIFI44L-1 的 mRNA 转录本显著下调,分别是 EGFP 组的 0.09 倍(p < 0.001)和 0.06 倍(p < 0.001)。这些结果表明,CgIFI44L-1 是牡蛎中一种保守的 ISG,受 CgIFNLP 和 CgSTAT 调节,参与牡蛎抗病毒免疫反应。

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