【作者】 刘冉阳； 史成银； 谢国驷； 王海波；

【Author】 LIU Ranyang;SHI Chengyin;Xie Guosi;WANG Haibo;Key Laboratory of Sustainable Development if Marine Fisheries, Ministry of Agriculture;Yellow Sea Fisheries Research Institute, Chinese Academy if Fishery Science;

【机构】 农业部海洋渔业可持续发展重点实验室黄海水产研究所中国水产科学研究院； 上海海洋大学水产与生命技术学院；

【摘要】 利用福尔马林灭活法制备了哈维氏弧菌疫苗,并以黄芪多糖作为佐剂,腹腔注射接种牙鲆?在第35天采取血样并检测抗体效价?于免疫后14天和35天取脾?头肾组织,提取mR NA,应用实时荧光定量PCR法检测2个组织中主要组织相容性复合体MHCⅠ和MHCⅡ?干扰素(IFN-γ)?白介素(IL)-6?白细胞表面受体(CD40)?Mx蛋白(Mx)和自然杀伤细胞增强因子(NKEF)这7种免疫相关基因的表达?第35天进行人工攻毒实验,测定免疫保护力。结果表明,黄芪多糖佐剂组和无佐剂组两组的抗体效价都在第35天达到2~7。定量PCR结果显示黄芪多糖佐剂疫苗引起各组织中7种基因表达显著上调,基因表达量最高值是对照组的2.2-32倍;黄芪多糖佐剂疫苗组基因的表达量显著高于无佐剂疫苗组?攻毒后黄芪多糖佐剂疫苗组的免疫保护率为80.2%,无佐剂疫苗组为74.4%?实验结果证明,哈维氏弧菌疫苗对牙鲆具有免疫作用,而且添加黄芪多糖佐剂组的免疫效果好于无佐剂组。更多还原

【Abstract】 The inactivated vaccine of pathogenic Vibrio harveyi was prepared by formalin,and then mixed with astragalan-adjuvant.Flounder Paralichthys olivaceus were vaccinated by intraperitoneal injection. Antibody titers were examined on the 35 th day after firstvaccination. The spleen and head kidney were taken at 14 th and 35 th day after immunization, respectively, and then total mR NA was extracted and reversely transcribed into cD NA.The expression of 7 immune related genes including the major histocompatibility complex class(MHCI)and(MHCII),Interferon(IFN-γ), interleukin-6(IL-6),cluster of differentiation 40(CD40),interferon-induced Myxovirus resistance protein(Mx), and natural killer cell enhancing factor(NKEF), genes was analyzed with quantitative real-time polymerase chain reaction(qR T-PCR) using β-actin as internal reference.Finally, all the fish were challenged with the bacteria(Vibrio harveyi) on 35 th day.There sults showed that the antibody titers of both groups also reached 2~7 on 35 th day. In astragalan-adjuvant group, the expression of all the tested genes was up-regulated significantly,2.2-32 folds higher than that of control.The transcriptional level of genes in livers,astragalan-adjuvant group was higher than that of in group whiout adjuvant.The RPS of astragalan-adjuvant group was 80.2%, and 74.4% in group whiout adjuvant.There sults suggest that vaccine can enhance the immunity in Paralichthys olivaceus.In addition, the immune enhancement was improved in astragalan-adjuvant group, indicating that astragalan-adjuvant has a positive effect on immunity.更多还原