乌鳢IL-1RAcP的分子特征及其对两种病原菌感染的免疫应答  

Molecular Characterization of IL-1RAcP in Snakehead(Channa argus)and Its Immune Response to the Infection with Two Pathogenic Bacteria

作　　者：李冬琦 崔正伟 赵飞[2] 邓玉婷 谭爱萍[2] 黄志斌[2] 姜兰[2] LI Dong-Qi;CUI Zheng-Wei;ZHAO Fei;DENG Yu-Ting;TAN Ai-Ping;HUANG Zhi-Bin;JIANG Lan(National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University,Shanghai 201306,China;Pearl River Fisheries Research Institute/Key Laboratory of Fishery Drug Development of Ministry of Agriculture and Rural Affairs/Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province,Chinese Academy of Fishery Sciences,Guangzhou 510380,China;Key Laboratory of Marine Biotechnology of Fujian Province/Institute of Oceanology,Fujian Agriculture and Forestry University,Fuzhou 350002,China)

机构地区：[1]上海海洋大学水产科学国家级实验教学示范中心,上海201306 [2]中国水产科学研究院珠江水产研究所/农业农村部渔用药物创制重点实验室/广东省水产动物免疫技术重点实验室,广州510380 [3]福建农林大学海洋研究院/福建省海洋生物技术重点实验室,福州350002

出　　处：《农业生物技术学报》2022年第1期96-105,共10页Journal of Agricultural Biotechnology

基　　金：中国水产科学研究院中央级公益性科研院所基本科研业务费专项(2021SJ-XT3);现代农业产业技术体系建设专项(CARS-46)。

摘　　要：白细胞介素l受体辅助蛋白(interleukin-1 receptor accessory protein, IL-1RAcP)是参与促炎细胞因子白细胞介素1 (interleukin-1, IL-1)信号传导的重要分子,在调节机体的免疫应答、炎症反应和抗菌感染等过程中发挥重要作用。为了探究鱼类IL-1RAcP在防御病原体感染过程中的作用机制,本研究克隆了乌鳢(Channa argus) IL-1RAcP基因shIL-1RAcP (GenBank No. MW928496),并初步分析了shIL-1RAcP对舒伯特气单胞菌(Aeromonas schubertii)和鰤诺卡氏菌(Nocardia seriolae) 2种病原菌感染及脂磷壁酸(lipoteichoic acid, LTA)、脂多糖(lipopolysaccharides, LPS)和聚肌苷酸胞苷酸(polyinosinic-polycytidylic acid, Poly(I:C)) 3种病原类似物和重组IL-1β蛋白刺激的免疫应答反应。结果显示,shIL-1RAcP基因的开放阅读框(ORF)长度为1 848 bp,预测编码615个氨基酸。编码蛋白的序列分析显示,sh IL-1RAcP具有IL-1家族成员典型的结构特征,含有3个胞外免疫球蛋白样(Ig-like)结构域、1个跨膜结构域和1个胞内Toll/IL-1受体(TIR)结构域。系统进化树分析显示,shIL-1RAcP与其他鱼类IL-1RAcP聚为一簇,并与大口黑鲈(Micropterus salmoides)和黄条鰤(Seriola lalandi) IL-1RAcP的同源性最近。qPCR检测显示,shIL-1RAcP在健康乌鳢的所有检测组织中广泛表达,其中在肝脏中的表达量最高,在头肾、体肾、脾脏等免疫组织中表达量较高,在心脏中的表达量最低。对2种病原菌感染的免疫应答分析显示,舒伯特气单胞菌和鰤诺卡氏菌能够诱导乌鳢体内头肾和脾脏中shIL-1RAcP的表达。LTA、LPS、Poly(I:C)和乌鳢的重组IL-1β蛋白(shIL-1β)能够诱导体外头肾白细胞中shIL-1RAcP的表达。本研究结果证实shIL-1RAcP在防御病原菌感染的免疫应答过程中发挥重要作用,为进一步揭示鱼类IL-1RAcP在抗菌免疫中的作用提供了基础资料。Interleukin-1 receptor accessory protein(IL-1RAcP) is an important molecule involved in the proinflammatory cytokine interleukin-1(IL-1) signal transduction, which plays critical roles in the inflammation,immune response and antimicrobial defense. To explore the mechanism of IL-1RAcP in the fish immune defense response to pathogen infection, the IL-1RAcP gene named shIL-1RAcP of snakehead(Channa argus)was cloned, and its immune response to the infection with 2 pathogenic bacteria and the stimulation with 3 pathogen analogues or recombinant IL-1β protein was preliminarily analyzed. The results showed that the fulllength ORF of shIL-1RAcP was 1 848 bp(GenBank No. MW928496), which encoded a putative protein of 615 amino acids. Sequences analysis showed that shIL-1RAcP possessed a typical structural characterization composed of three extracellular Ig-like domains, one transmembrane domain, and one intracellular Toll/IL-1 receptor(TIR) domain. The phylogenetic tree analysis revealed that shIL-1RAcP clustered within a subgroup of other teleost species and shared the closest evolutionary relationship with the IL-1RAcP proteins of Micropterus salmoides and Seriola lalandi. Moreover, qPCR analysis showed that shIL-1RAcP was widely expressed with different expression levels in all tested tissues of healthy snakehead. The highest expression of shIL-1RAcP was in the liver, followed by several immune tissues, including the head kidney, spleen, and truck kidney, whereas the lowest expression was detected in the heart. After infection with Nocardia seriolae or Aeromonas schubertii, the relative expressions of shIL-1RAcP were mainly up-regulated in the head kidney and spleen in vivo at different time points, respectively. Similarly, after stimulation with three pathogen analogues and recombinant shIL-1β protein, the expressions of shIL-1RAcP in head kidney leukocytes in vitro could be induced by lipoteichoic acid(LTA), lipopolysaccharides(LPS), polyinosinic-polycytidylic acid [Poly(I: C)],and recombinant shIL-1β protein. In c

关 键 词：乌鳢 白细胞介素l受体辅助蛋白(IL-1RAcP) 舒伯特气单胞菌 鰤诺卡氏菌 免疫应答 

分 类 号：S917.4[农业科学—水产科学]