Department of Marine Biology, College of Oceanography, Hohai University, 1 Xikang Road, Nanjing, Jiangsu, 210098, China.
Biodiversity and Biosafety Research Center, Nanjing Institute of Environmental Sciences, 8 Jiangwangmiao Road, Nanjing, Jiangsu, 210042, China.
Dev Comp Immunol. 2022 Sep;134:104456. doi: 10.1016/j.dci.2022.104456. Epub 2022 May 28.
Tumor necrosis factor (TNF) is one of the most important cytokines involved in various biological processes in vertebrates and invertebrates. In the present study, a new member of the TNF superfamily (named EsTNFSF) was identified from the Chinese mitten crab (Eriocheir sinensis). The full-length cDNA of EsTNFSF is 2462 bp and encodes a polypeptide with 499 amino acids. The deduced EsTNFSF protein contained a transmembrane region and a conserved extracellular C-terminal TNF domain. Phylogenetic analysis indicated that EsTNFSF was closely related to other TNFSFs from crustaceans. Quantitative real-time PCR analysis showed that EsTNFSF was expressed in all the tissues examined, and the highest expression was found in the hepatopancreas. The mRNA levels of EsTNFSF in hemocytes underwent a time-dependent and variable degree of enhancement after stimulation with lipopolysaccharide, peptidoglycan, Staphylococcus aureus, and Vibrio parahaemolyticus. Functionally, EsTNFSF knockdown by siRNA suppressed the transcriptional levels of c-Jun N-terminal kinase and two antimicrobial peptides, anti-lipopolysaccharide factor and crustin. Furthermore, purified recombinant EsTNFSF protein accelerated the bacterial clearance in vivo and inhibited the growth of V. parahaemolyticus and S. aureus in vitro. The results revealed that EsTNFSF, as an inducible immune response gene, plays a crucial role in the antibacterial immune defense of E. sinensis.
肿瘤坏死因子（TNF）是参与脊椎动物和无脊椎动物各种生物过程的最重要细胞因子之一。本研究从中华绒螯蟹（Eriocheir sinensis）中鉴定出一种 TNF 超家族的新成员（命名为 EsTNFSF）。EsTNFSF 的全长 cDNA 为 2462bp，编码一个含有 499 个氨基酸的多肽。推断的 EsTNFSF 蛋白含有跨膜区和保守的细胞外 C 端 TNF 结构域。系统发育分析表明，EsTNFSF 与其他甲壳类动物的 TNFSFs 密切相关。定量实时 PCR 分析显示，EsTNFSF 在所有检测到的组织中均有表达，在肝胰腺中表达量最高。EsTNFSF 在血细胞中的 mRNA 水平在受到脂多糖、肽聚糖、金黄色葡萄球菌和副溶血弧菌刺激后呈时间依赖性和不同程度的增强。功能上，siRNA 介导的 EsTNFSF 敲低抑制了 c-Jun N 端激酶和两种抗菌肽（抗脂多糖因子和 crustin）的转录水平。此外，纯化的重组 EsTNFSF 蛋白在体内加速了细菌清除，并在体外抑制了副溶血弧菌和金黄色葡萄球菌的生长。结果表明，EsTNFSF 作为一种诱导免疫反应基因，在中华绒螯蟹的抗菌免疫防御中发挥着关键作用。
