Aquaculture International ( IF 2.9 ) Pub Date : 2021-07-26 , DOI: 10.1007/s10499-021-00721-6 Sweta Das 1 , Arathi Dharmaratnam 1 , Charan Ravi 1 , Raj Kumar 1 , Thangaraj Raja Swaminathan 1
Goldfish is one of the preferred ornamental fish which is highly susceptible to cyprinid herpesvirus-2 (CyHV-2) infection. The present study aimed to analyse immune gene expression in a co-culture of CyHV-2-sensitized goldfish peripheral blood leukocytes (PBLs) with CyHV-2-infected fantail goldfish fin cell lines (FtGF). Goldfish were sensitized with intraperitoneal TCID50 dose (107.8±0.26/mL) of CyHV-2. After 2 weeks, PBLs were collected and co-cultured with CyHV-2-infected FtGF cells keeping both uninfected FtGF cells and PBL control groups. After 2 days of co-culture, WST-1 assay for cell proliferation was performed at 450 nm during the 2nd, 4th and 6th days of co-culture. The results showed a significant increase (p < 0.05) in cell density in CyHV-2-infected PBL and virus-infected FtGF cells during the 4th day post co-culture which confirmed effector cell generation. Expressions of few immune genes were checked taking RNA samples of CyHV-2-induced PBLs post co-culture with infected FtGF cells along with uninfected FtGF cells as control group at different time periods (2nd, 4th and 6th days) in triplicate. The results indicated increased expression of CD8α, IFNγ, b2m, MHC I, LMP 7, IL-10, IL-12 and GATA3 except Tapasin. From the above study, we concluded that goldfish showed both Th1- and Th2-mediated immune responses to CyHV-2. The current findings support the scope for further vaccine development against CyHV-2 for goldfish.
中文翻译:
与 CyHV-2 感染的金鱼鳍细胞系共培养的 cyprinid herpesvirus-2 (CyHV-2) 致敏外周血白细胞 (PBL) 中的免疫基因表达
金鱼是首选的观赏鱼之一,对鲤科疱疹病毒2(CyHV-2)感染高度敏感。本研究旨在分析 CyHV-2 致敏金鱼外周血白细胞 (PBL) 与 CyHV-2 感染扇尾金鱼鳍细胞系 (FtGF) 共培养中的免疫基因表达。金鱼用腹腔内 TCID 50剂量 (10 7.8±0.26/mL) CyHV-2。2 周后,收集 PBLs 并与 CyHV-2 感染的 FtGF 细胞共培养,保留未感染的 FtGF 细胞和 PBL 对照组。共培养 2 天后,在共培养的第 2、第 4 和第 6 天在 450 nm 处进行细胞增殖的 WST-1 测定。结果显示,在共培养后第 4 天,CyHV-2 感染的 PBL 和病毒感染的 FtGF 细胞的细胞密度显着增加 (p < 0.05),这证实了效应细胞的产生。在不同时间段(第 2、第 4 和第 6 天)以 CyHV-2 诱导的 PBL 与感染的 FtGF 细胞以及未感染的 FtGF 细胞共培养后的 RNA 样品作为对照组,检查少数免疫基因的表达,一式三份。结果表明除了 Tapasin 外,CD8α、IFNγ、b2m、MHC I、LMP 7、IL-10、IL-12 和 GATA3 的表达增加。从上述研究中,我们得出结论,金鱼对 CyHV-2 表现出 Th1 和 Th2 介导的免疫反应。目前的研究结果支持进一步开发针对金鱼的 CyHV-2 疫苗的范围。