Guangzhou Key Laboratory of Subtropical Biodiversity and Biomonitoring, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, College of Life Science, South China Normal University, Guangzhou, 510631, PR China; State Key Laboratory of Developmental Biology of Freshwater Fish, College of Life Science, Hunan Normal University, Changsha, 410081, PR China.
Guangzhou Key Laboratory of Subtropical Biodiversity and Biomonitoring, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, College of Life Science, South China Normal University, Guangzhou, 510631, PR China.
Fish Shellfish Immunol. 2019 Jun;89:486-497. doi: 10.1016/j.fsi.2019.04.020. Epub 2019 Apr 11.
CD59, a multifunctional glycoprotein, not only plays a regulatory role in complement cascades, but also participates in modulation of teleostean immunity. In this study, full length sequence of EcCD59 was obtained, comprising a 5'UTR of 163 bp, an ORF of 354 bp and a 3'UTR of 559 bp. EcCD59 gene encoded a polypeptide of 117 amino acids. Tissue-specific analysis revealed that the highest expression of EcCD59 mRNA was observed in muscle. Vibrio alginolyticus challenge can significantly increase EcCD59 mRNA expression in liver, kidney and spleen. EcCD59 distribution was detected by a combined approach using GFP-overexpression, immunofluorescence and ELISA assay, indicating that EcCD59 may be predominantly aggregated in cellular membrane. Both EcCD59 and EcCD59delGPI can directly bind to V. alginolyticus and decrease the in vitro growth of V. alginolyticus. Additionally, vibrio injection experiment indicated that the binding of EcCD59 or EcCD59delGPI to V. alginolyticus can restrict its growth rate in vivo. In this study, we found that EcCD59 may be involved in immune defense against vibrio infection in a complement-independent manner.
CD59 是一种多功能糖蛋白,不仅在补体级联反应中发挥调节作用,还参与调节鱼类的免疫。本研究获得了全长 EcCD59 序列,包括 163bp 的 5'UTR、354bp 的 ORF 和 559bp 的 3'UTR。EcCD59 基因编码一个由 117 个氨基酸组成的多肽。组织特异性分析显示 EcCD59 mRNA 的表达量在肌肉中最高。哈维弧菌刺激可显著增加肝、肾和脾中 EcCD59 mRNA 的表达。通过 GFP 过表达、免疫荧光和 ELISA 检测的联合方法检测 EcCD59 的分布,表明 EcCD59 可能主要聚集在细胞膜上。EcCD59 和 EcCD59delGPI 均可直接与哈维弧菌结合,降低哈维弧菌的体外生长速度。此外,弧菌注射实验表明 EcCD59 或 EcCD59delGPI 与哈维弧菌的结合可以限制其在体内的生长速度。本研究发现,EcCD59 可能通过非补体依赖的方式参与鱼类抵抗弧菌感染的免疫防御。