College of Fishery, Guangdong Ocean University, Zhanjiang, 524088, China; Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals, Zhanjiang, 524088, China; Guangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals, Zhanjiang 524088, China.
College of Fishery, Guangdong Ocean University, Zhanjiang, 524088, China; Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals, Zhanjiang, 524088, China; Guangdong Key Laboratory of Control for Diseases of Aquatic Economic Animals, Zhanjiang 524088, China.
Fish Shellfish Immunol. 2018 Jan;72:259-265. doi: 10.1016/j.fsi.2017.11.002. Epub 2017 Nov 5.
NK-lysin, an effector of cytotoxic T cells and natural killer cells, is a potent antimicrobial peptide widely distributed in mammals. Homologues of NK-lysin have been discovered in several teleost species, but only several of their natural functions was recorded so far. Here we identified an NK-lysin from Nile tilapia (Oreochromis niloticus), On-NKL, and analysed its expression model and biological effects on pathogen infection. The open reading frame of On-NKL sequence spans 432 bp, codes for 143 amino acids and shares 27%-62% overall sequence identities with NK-lysin of other species. The deduced mature peptide of On-NKL possesses a saposin B domain and six well-conserved cysteine residues that essential for antimicrobial activity by forming three intrachain disulphide bonds. The results of qRT-PCR showed that On-NKL expression was observed in multiple tissues and head kidney leucocytes and nonspecific cytotoxic cells (NCCs) and is most abundant in gills. After bacterial challenge, On-NKL expression significantly varied in different tissues and NCCs. Following bacterial infection, On-NKL-overexpressing fish featured significantly lower pathogen loads in tissues than control fish. On-NKL-overexpressing fish also exhibited 33.3% relative percent survival compare with control groups. Findings suggested that On-NKL could be the potential effector of NCCs and act as immune-related gene that enhances antimicrobial defence.
NK 细胞溶素是细胞毒性 T 细胞和自然杀伤细胞的效应因子,是一种广泛分布于哺乳动物中的强效抗菌肽。NK 细胞溶素的同源物已在几种硬骨鱼物种中被发现,但迄今为止仅记录了它们的一些天然功能。在这里,我们从尼罗罗非鱼(Oreochromis niloticus)中鉴定出一种 NK 细胞溶素,命名为 On-NKL,并分析了其表达模式及其对病原体感染的生物学效应。On-NKL 序列的开放阅读框跨度为 432bp,编码 143 个氨基酸,与其他物种的 NK 细胞溶素的整体序列同一性为 27%-62%。On-NKL 的推断成熟肽具有一个 Saposin B 结构域和六个保守的半胱氨酸残基,这些残基对于通过形成三个链内二硫键发挥抗菌活性至关重要。qRT-PCR 的结果表明,On-NKL 在多种组织和头肾白细胞及非特异性细胞毒性细胞(NCC)中均有表达,在鳃中表达最为丰富。在细菌攻毒后,On-NKL 在不同组织和 NCC 中的表达显著变化。在细菌感染后,过表达 On-NKL 的鱼类组织中的病原体载量明显低于对照组鱼类。过表达 On-NKL 的鱼类的相对存活百分比也比对照组高 33.3%。这些发现表明,On-NKL 可能是 NCC 的潜在效应因子,并作为增强抗菌防御的免疫相关基因发挥作用。