【作者】 王雪鹏； 宋憬愚； 丁雷； 李福昌；

【Author】 Xuepeng Wang, Jingyu Song, Lei Ding, Fuchang Li (College of Animal Science and Technology, Shandong Agricultural University, Taian 271018)

【机构】 山东农业大学动物科技学院；

【摘要】 本研究成功构建溶菌质粒载体,并将其转入迟缓爱德华菌,通过温度诱导制备出迟缓爱德华菌菌蜕疫苗。分别用1×105cells/ml(A组)、1×106cells/ml(B组)、1×107cells/ml(C组)的迟缓爱德华菌菌蜕疫苗和PBS(D组)免疫黑鲷30尾,在免疫后的第0、3、7、14、21和28d采样,并测定血清抗体水平和白细胞吞噬活性。结果表明:B和C组血清抗体水平显著高于A组(P<0.05)和对照组(P<0.01),B和C组间无显著差异(P>0.05);在免疫3d后,实验组的吞噬百分比(PP)与对照组相比均有显著差异(P<0.05),随着时间的延长,各实验组PP继续升高;在免疫后第21d,105,106和107这3个不同浓度的实验组PP均达到最高值,分别为45.7%,51.2%和50.7%;吞噬指数(PI)的变化略有不同,免疫7d后显著高于对照组水平(P<0.05),第14d3个实验组PI均达到最高水平。结果显示迟缓爱德华菌菌蜕疫苗能诱导强烈的细胞免疫和体液免疫,并有效地激活机体的免疫保护应答,是一种理想的疫苗体系。更多还原

【Abstract】 Bacterial ghosts may be generated by the controlled expression of the PhiX174 lysis gene E in Gram negative bacteria and they are intriguing vaccine candidates since ghosts retain functional antigenic cellular determinants often lost during traditional inactivation procedures. The Edwardsiella tarda ghost (ETG) vaccine was prepared using this technology and tested in vaccination trials in this study. Fish were divided into three immunization groups (A to C groups were respectively immunized with ETG at 1×105cells/ml, 1×106cells/ml, 1×107cells/ml, n=30 for each group) or fish (D, control group) treated with phosphate-buffered saline (PBS), respectively. Immune responses were measured after 0, 3, 7, 14, 21, 28 days of treating. The results showed that serum antibody titers were significantly higher in B and C groups compared to A (P<0.05)and D (P<0.01)groups, while there was no significantly difference in B and C groups(P>0.05). Phagocytic activity (percent phagocytes, PP) was significantly higher (p<0.05) in ETG immunized groups than in the control group after treated 3 days, and the PP continues to rise with time until day 21, were 45.7%,51.2% and 50.7%, respectively. However, the phagocytic activity (phagocytic index) was different, which was significantly higher (p<0.05) in ETG immunized groups than in the control group after treated 7 days, and were highest on the day 14. All these results suggested that an ETG could stimulate cellular, mucosal and humoral immunity, and could be used as vaccine candidates.更多还原