摘要
为深入研究羊口疮疫苗诱导机体产生免疫效应的分子机理,用羊口疮疫苗免疫兔,并对其血清中的特异性抗体水平和外周血免疫相关细胞因子的表达水平进行研究。将500 m L灭活的羊口疮病毒感染细胞毒液和经超声波破碎的灭活羊口疮病毒感染细胞毒液分别浓缩至100 m L,按照佐剂说明书比例,分别与弗氏佐剂、201VG、1313VG、GEL01、11R VG混匀。另外,取灭活的羊口疮痂皮毒液10 m L与201 VG混匀,制备成疫苗。用所制备的不同佐剂疫苗各免疫2只兔子。在免疫后不同时间采集外周血,应用琼脂免疫扩散方法检测血清中特异性抗体效价,同时应用实时荧光定量PCR方法检测外周血中细胞因子表达水平。结果显示:在兔子接种羊口疮疫苗后期,血清中抗体效价除了"1313VG+未破碎病毒"组为1:8之外,其他组别的抗体效价均达到1:16或1:32;各组别外周血中Th1型细胞因子(IL-2、IFN-γ、TNF-α)的表达量与对照组相比总体上升,而Th2型细胞因子(IL-4、IL-6)的表达量总体呈下降趋势。结果表明,羊口疮疫苗可以同时诱导机体产生细胞免疫应答和体液免疫应答,但对该两种免疫应答的诱导水平不同。
In order to better understand the molecular mechanism of immune response induced by sheep ecthyma vaccine, the level of specific antibodies in serum and expression of related cytokines in peripheral blood of rabbits immunized with sheep ecthyma vaccine were detected. 500 mL inactivated sheep ecthyma virus-infected cell suspensions were condensed into 100 mL, as well as another 500 mL suspensions with ultrasonic fragmentation treatment. Then the two portions of concentrated solutions were respectively mixed with Freund＇s adjuvant, 201 VG, 1313 VG, GEL01 and llR VG to prepare different vaccines. In addition, 10 mL inactivated sheep ecthyma crust venom was also used to generate vaccine by being mixed with 201 VG. Each kind of vaccine was applied to immunize two rabbits, and then blood samples were collected at different time to detect the level of specific antibodies in serum by agar immunodiffusion assay and the expression of cytokines in peripheral blood by real-time fluorescence quantitative PCR. The results showed that the serum antibody titer all reached 1：16 or 1：32 in later period of immunization except the vaccine ＂1313VG＋inactivated virus-infected cell suspensions＂, antibody titer of which was 1：8. Interestingly, the expression quantity of Thl cytokines （IL-2, IFN- y and TNF- a ） in peripheral blood of all experimented rabbits increased in comparison with the control group. By contrast, the expression ofTh2 cytokines （IL-4, IL-6） decreased. In conclusion, both humoral immune response and cellular immune response of rabbits were induced by sheep ecthyma vaccines prepared, and the changes of the two kinds of immune responses varied.
出处
《中国动物检疫》
CAS
2016年第11期89-93,107,共6页
China Animal Health Inspection
关键词
羊口疮疫苗
体液免疫
细胞免疫
特异性抗体
细胞因子
佐剂
sheep ecthyma vaccine
humoral immune response
cellular immune response
specific antibody
cytokine
adjuvant
