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大黄鱼虹彩病毒灭活疫苗对斑马鱼的注射免疫效果

Injection immunization effect of inactivated large yellow croaker iridovirus vaccine on zebrafish

  • 摘要: 为了研究大黄鱼虹彩病毒(large yellow croaker iridovirus, LYCIV)灭活疫苗注射免疫效果,将斑马鱼(Danio rerio)分为4组(对照组、灭活病毒组、佐剂组、疫苗组),分别注射细胞培养液、灭活LYCIV、佐剂、灭活疫苗。免疫后第21天,每尾斑马鱼腹腔注射LYCIV (5×107 copies/尾),采用荧光定量PCR法测定免疫后第10、21、35天脾、肾组织免疫相关基因ifngil1bil12ail2mxirf1tlr9myd88tnfa的表达水平。结果表明,对照组、灭活病毒组、佐剂组和疫苗组攻毒后的存活率分别为14.29%、28.57%、28.57%和42.86%,疫苗组存活率显著高于对照组(P<0.05),相对免疫保护率为33.33%;疫苗组中脾组织ifngil1bil12amxirf1tlr9myd88tnfa在第21天的表达水平极显著高于对照组、灭活病毒组和佐剂组(P<0.01);疫苗组中肾组织ifngil1bil12amxirf1tlr9tnfa在第21天的表达水平极显著高于对照组、灭活病毒组和佐剂组(P<0.01)。研究表明,以Montanide TM GEL 02 PR为佐剂制备的LYCIV灭活疫苗能提高斑马鱼IFN-γ、IRF1、TLR9、MyD88等干扰素相关信号分子和IL-1β等炎症因子的表达水平,提高斑马鱼感染LYCIV后的存活率。

     

    Abstract: To study the immunological effect of inactivated large yellow croaker iridovirus (LYCIV) vaccine on zebrafish (Danio rerio), the fishes tested were divided into 4 groups (control group, inactivated virus group, adjuvant group, and vaccine group) and injected with cell culture solution, inactivated LYCIV, adjuvant and inactivated vaccine respectively. On the 21d post-immunization, each zebrafish was intraperitoneally injected with LYCIV at 5×107 copies/fish. The expression levels of immune-related genes ifng, il1b, il12a, il2, mx, irf1, tlr9, myd88 and tnfa in the spleen and kidney tissues were determined by real-time quantitative PCR in 10 d, 21 dfand 35 d post-immunization. Results showed that the survival rates of control group, inactivated virus group, adjuvant group and vaccine group were 14.29%, 28.57%, 28.57%, and 42.86%, respectively. The survival rate of the vaccine group was significantly higher than that of the control group (P<0.05) with a survival of 33.33 %. The genes ifng, il1b, il12a, mx, irf1, tlr9, myd88 and tnfa in the spleen tissue of vaccine group had significantly higher expression than those of the control, inactivated virus and adjuvant groups (P<0.01). The expression of the genes ifng, il1b, il12a, mx, irf1, tlr9 and tnfa in kidney tissues of the fish in vaccine group were significantly higher than those in the control, inactivated virus and adjuvant groups on day 21 after immunization (P<0.01). It indicates that the LYCIV inactivated vaccine prepared with MontanideTM GEL 02 PR as adjuvant could improve the expression of interferon-related signaling molecules such as IFN-γ, IRF1, TLR9, MyD88 and inflammatory factors such as IL-1β in zebrafish besides the survival rate after infection.

     

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