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日本沼虾 2-跨膜 C 型凝集素参与抗菌免疫反应。

2-Transmembrane C-type lectin from oriental river prawn Macrobrachium nipponense participates in antibacterial immune response.

机构信息

College of Oceanography, Hohai University, 1 Xikang Road, Nanjing, Jiangsu, 210098, China; Postdoctoral Innovation Practice Base, Jiangsu Shuixian Industrial Company Limited, 40 Tonghu Road, Baoying, Yangzhou, Jiangsu, 225800, China.

College of Marine Science and Engineering, Nanjing Normal University, 1 Wenyuan Road, Nanjing, Jiangsu, 210023, China.

出版信息

Fish Shellfish Immunol. 2019 Aug;91:58-67. doi: 10.1016/j.fsi.2019.05.029. Epub 2019 May 13.

Abstract

As a type of pattern-recognition proteins (PRRs), C-type lectins (CTLs) perform important functions in non-self recognition and clearance of pathogens in innate immunity. In this study, a unique 2-transmembrane CTL (designated as Mn-2TM-cLec) with a single carbohydrate recognition domain (CRD) was isolated from Macrobrachium nipponense. The full-length cDNA of Mn-2TM-cLec consisted of 3265 bp with an 837 bp open reading frame encoding a protein with 278 amino acids. Mn-2TM-cLec was ubiquitously distributed in various tissues of normal prawn, particularly in the hemocytes, hepatopancreas, and gills. The expression of Mn-2TM-cLec was significantly up-regulated in the gills and hepatopancreas after the prawns were challenged with Staphylococcus aureus and Vibrio parahaemolyticus. RNA interference knock-down of Mn-2TM-cLec gene decreased the transcription levels of three antimicrobial peptides (anti-lipopolysaccharide factor (ALF) 1, ALF2, and Crustin (Crus) 1) after V. parahaemolyticus infection. The recombinant CRD of Mn-2TM-cLec could bind lipopolysaccharide, peptidoglycans, and diverse bacterial strains and agglutinate S. aureus and V. parahaemolyticus in a Ca-dependent manner. In addition, the rCRD enhanced the clearance of V. parahaemolyticus injected in prawns. In summary, Mn-2TM-cLec might act as a PRR to participate in the prawn immune defense against pathogens through its antimicrobial activity.

摘要

作为一种模式识别蛋白(PRRs),C 型凝集素(CTLs)在先天免疫中对非自身的识别和清除病原体起着重要作用。本研究从日本沼虾(Macrobrachium nipponense)中分离出一种具有独特的 2 次跨膜 CTL(命名为 Mn-2TM-cLec),它具有单个碳水化合物识别结构域(CRD)。Mn-2TM-cLec 的全长 cDNA 长 3265bp,包含一个 837bp 的开放阅读框,编码一个由 278 个氨基酸组成的蛋白质。Mn-2TM-cLec 在正常虾的各种组织中广泛分布,特别是在血细胞、肝胰腺和鳃中。虾受到金黄色葡萄球菌和副溶血弧菌攻击后,Mn-2TM-cLec 在鳃和肝胰腺中的表达显著上调。Mn-2TM-cLec 基因的 RNA 干扰敲低降低了副溶血弧菌感染后三种抗菌肽(抗脂多糖因子(ALF)1、ALF2 和 Crustin(Crus)1)的转录水平。Mn-2TM-cLec 的重组 CRD 可以与脂多糖、肽聚糖和多种细菌菌株结合,并以 Ca 依赖性方式凝集金黄色葡萄球菌和副溶血弧菌。此外,rCRD 增强了注射在虾体内的副溶血弧菌的清除。综上所述,Mn-2TM-cLec 可能作为一种 PRR,通过其抗菌活性参与虾对病原体的免疫防御。

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