在这项研究中,建立了一种新的细胞系日本比目鱼脾脏(JFSP),该细胞系源自日本比目鱼(Paralichthysolivaceus)的脾脏。JFSP细胞在29℃生长迅速,L-15培养基中的最佳胎牛血清浓度为15%。细胞传代培养超过80代。JFSP细胞的二倍体染色体数为2n=68,与正常二倍体日本比目鱼的染色体数不同。建立的细胞对Bohle病毒(BIV)易感,病毒性出血性败血症病毒(VHSV),Hirame弹状病毒(HIRRV),传染性造血系统坏死病毒(IHNV),和淋巴细胞病病毒(LCDV),不同程度的细胞病变效应(CPE)证明了这一点。通过qRT-PCR和透射电子显微镜确认病毒在JFSP细胞中的复制。此外,四个免疫相关基因的表达,TRAF3,IL-1β,TNF-α,和TLR2在病毒感染后差异改变。结果表明,细胞经历了抗病毒免疫反应。JFSP细胞系是病毒学的理想体外工具。利用鱼类细胞系研究鱼类的免疫基因和免疫机制,阐明鱼类的免疫机制,对于从根本上预防和治疗鱼类疾病具有重要的理论意义和实际应用价值。
A new cell line Japanese flounder spleen (JFSP) derived from the spleen of Japanese flounder (Paralichthys olivaceus) was established and characterized in this
study. The JFSP cells grew rapidly at 29 °C, and the optimum fetal bovine serum concentration in the L-15 medium was 15%. Cells were subcultured for more than 80 passages. The JFSP cells have a diploid chromosome number of 2n = 68, which differs from the chromosome number of normal diploid Japanese flounder. The established cells were susceptible to Bohle virus (BIV), Viral hemorrhagic septicemia virus (VHSV), Hirame rhabdovirus (HIRRV), Infectious hematopoietic necrosis virus (IHNV), and Lymphocystis disease virus (LCDV), as evidenced by varying degrees of cytopathic effects (CPE). Replication of the virus in JFSP cells was confirmed by qRT-PCR and transmission electron microscopy. In addition, the expression of four immune-related genes, TRAF3, IL-1β, TNF-α, and TLR2, was differentially altered following viral infection. The results indicated that the cells underwent an antiviral immune response. JFSP cell line is an ideal tool in vitro for virology. The use of fish cell lines to
study the immune genes and immune mechanism of fish and to clarify the immune mechanism of fish has important theoretical significance and practical application value for the fundamental prevention and treatment of fish diseases.