Marine Science and Engineering College, Qingdao Agricultural University, Qingdao, 266109, China.
Marine Science and Engineering College, Qingdao Agricultural University, Qingdao, 266109, China.
Fish Shellfish Immunol. 2020 May;100:161-170. doi: 10.1016/j.fsi.2020.02.063. Epub 2020 Mar 2.
Chemokines are small cytokines that are classified into four groups, one of which is called CC chemokines. In the present study, the full-length cDNA of a CCL25 chemokine was identified from black rockfish, Sebastes schlegelii (named as SsCCL25) by EST (expressed sequence tag) analysis. The cDNA of SsCCL25 consisted of a 5-terminal untranslated region (UTR) of 74 bp, a 3-UTR of 882 bp with a poly (A) tail, and an open reading frame (ORF) of 303 bp encoding a polypeptide of 100 amino acids with the putative molecular mass of 11.5 kDa. There was a SCY domain in the deduced amino acid sequence of SsCCL25. The phylogenetic relationships and syntenic analyses provided evidences for the identities of SsCCL25 with CCL25 group. The mRNA transcripts of SsCCL25 were expressed in all detected tissues and dominantly in liver, muscle and gill. Moreover, after Vibrio anguillarum stimulation, the mRNA expression levels of SsCCL25 were significantly up-regulated at 24 h (p < 0.05) in the liver and during 4-8 h (p < 0.05) in the spleen. Recombinant SsCCL25 protein induced chemotaxis of both control and LPS-stimulated peripheral blood leukocytes (PBL) and enhanced their resistance to bacterial infection in a dose-dependent manner. Furthermore, rSsCCL25 showed significant inhibitory effect on V. anguillarum and Edwardsiella tarda growth. All these results collectively indicated that SsCCL25 might contribute to the defense against microbe infection and function as a chemoattractant in black rockfish.
趋化因子是一类小分子细胞因子,可分为四个亚群,其中之一被称为 CC 趋化因子。本研究通过 EST(表达序列标签)分析,从青石斑鱼(Sebastes schlegelii)中鉴定出一种 CCL25 趋化因子的全长 cDNA(命名为 SsCCL25)。SsCCL25 的 cDNA 由 74bp 的 5'非翻译区(UTR)、882bp 的 3'UTR 组成,带有 poly(A)尾,开放阅读框(ORF)编码一个 303bp 的多肽,包含 100 个氨基酸,推测分子量为 11.5 kDa。在 SsCCL25 的推导氨基酸序列中存在一个 SCY 结构域。系统发育关系和基因同线性分析为 SsCCL25 与 CCL25 亚群的同源性提供了证据。在所有检测的组织中均有 SsCCL25 的 mRNA 转录本表达,在肝脏、肌肉和鳃中表达丰度最高。此外,在鳗弧菌刺激后,SsCCL25 的 mRNA 表达水平在肝脏中于 24 小时(p<0.05)显著上调,在脾脏中于 4-8 小时(p<0.05)显著上调。重组 SsCCL25 蛋白可诱导对照组和 LPS 刺激的外周血白细胞(PBL)的趋化作用,并呈剂量依赖性增强其对细菌感染的抵抗力。此外,rSsCCL25 对鳗弧菌和迟缓爱德华氏菌的生长具有显著的抑制作用。所有这些结果表明,SsCCL25 可能有助于抵抗微生物感染,并作为青石斑鱼的趋化因子发挥作用。