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分子克隆、鉴定与表达分析的主要组织相容性复合体 I 类 alpha 基因的河豚(Takifugu obscurus)。

Molecular cloning, characterization and expression analysis of major histocompatibility complex class I alpha gene of pufferfish (Takifugu obscurus).

机构信息

Guangzhou Key Laboratory of Subtropical Biodiversity and Biomonitoring, School of Life Sciences, South China Normal University, Guangzhou, China.

Guangdong South China Sea Key Laboratory of Aquaculture for Aquatic Economic Animals, Guangdong Ocean University, Zhanjiang, China.

出版信息

J Fish Dis. 2021 May;44(5):613-625. doi: 10.1111/jfd.13307. Epub 2020 Nov 21.

Abstract

The major histocompatibility complex (MHC) genes play a key role in immune response in vertebrates. In this study, an MHC I alpha homolog gene (PfMHC Ⅰα) from pufferfish (Takifugu obscurus) was identified and its subcellular localization and expression patterns of PfMHC Ⅰα after challenge in vivo and in vitro were analysed. The open reading frame of PfMHC Ⅰα was 1,089 bp in length, encoding 362 aa. The immunofluorescence result revealed that PfMHC Ⅰα was presented on the membrane of lymphocytes. qRT-PCR analysis indicated that PfMHC Ⅰα was expressed in all examined tissues, with the highest expression in skin, followed by the expression in gills and whole blood. After challenge of Aeromonas hydrophila or polyinosinic: polycytidylic acid (Poly I:C) in vitro, the expression levels of PfMHC Ⅰα on pufferfish kidney lymphocytes were significantly up-regulated, with the highest expression level at 48 hr post-challenge. After infection with A. hydrophila or Poly I:C in vivo, the expression levels of PfMHC Ⅰα in the skin, whole blood and kidneys were significantly up-regulated. Taken together, it is speculated that PfMHC Ⅰα associates with resistance to both intracellular and extracellular antigens and plays an important role in the host response against pathogen infection in pufferfish.

摘要

主要组织相容性复合体 (MHC) 基因在脊椎动物的免疫反应中发挥关键作用。本研究从河豚鱼 (Takifugu obscurus) 中鉴定出 MHC I alpha 同源基因 (PfMHC Ⅰα),并分析了 PfMHC Ⅰα的亚细胞定位及其在体内和体外受到刺激后的表达模式。PfMHC Ⅰα 的开放阅读框长 1089bp,编码 362 个氨基酸。免疫荧光结果显示 PfMHC Ⅰα 呈现在淋巴细胞的膜上。qRT-PCR 分析表明 PfMHC Ⅰα 在所有检测组织中均有表达,在皮肤中的表达最高,其次是在鳃和全血中的表达。在体外受到嗜水气单胞菌或聚肌胞苷酸 (Poly I:C) 刺激后,河豚鱼肾淋巴细胞中 PfMHC Ⅰα 的表达水平显著上调,在刺激后 48 小时达到最高表达水平。在体内感染嗜水气单胞菌或 Poly I:C 后,皮肤、全血和肾脏中 PfMHC Ⅰα 的表达水平显著上调。综上所述,推测 PfMHC Ⅰα 与抵抗细胞内和细胞外抗原有关,在河豚鱼抵抗病原体感染的宿主反应中发挥重要作用。

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