羊口疮病毒F1L与B2L基因核酸疫苗联合免疫小鼠特性的研究

为评价羊口疮病毒(OrfV)F1L与B2L基因核酸疫苗PVAX1-F1L、PVAX1-B2L联合免疫小鼠诱导体液和细胞免疫的应答效果,分别将PVAX1-F1L+PVAX1-B2L、PVAX1-B2L、PVAX1-F1L、PVAX1空载体与PBS通过后腿肌肉注射方式免疫小鼠,采用ELISA方法检测免疫不同时间段的小鼠血清中OrfV特异性抗体及细胞因子IL-2、IL-4和IFN-γ,并通过MTT法检测免疫小鼠脾淋巴细胞增殖反应。结果显示,PVAX1-F1L+PVAX1-B2L联合免疫组小鼠血清抗体效价及IL-2、IFN-γ细胞因子水平显著(P<0.05)高于PVAX1-F1L和PVAX1-B2L单独免疫组,IL-4细胞因子水平差异不明显,但均与PVAX1空载体和PBS组差异极显著(P<0.01);PVAX1-F1L+PVAX1-B2L联合免疫组小鼠脾淋巴细胞增殖水平与PVAX1-F1L和PVAX1-B2L单独免疫组相比,差异显著(P<0.05),与PVAX1空载体和PBS组相比,差异极显著(P<0.01)。结果表明,PVAX1-F1L+PVAX1-B2L联合免疫小鼠能够增强OrfV特异性抗体的分泌和细胞免疫应答,这为羊口疮新型疫苗的研制奠定了理论基础。

Study on immune response of DNA vaccine consisted with F1L and B2L gene in mice

To evaluate the effect of inducing humoral and cellular immune response in mice immunized with DNA vaccines PVAX1-F1L and PVAX1-B2L of OrfV F1L and B2L gene,the test was divided into five groups PVAX1-F1L +PVAX1-B2L,PVAX1-B2L,PVAX1-F1L,PVAX1 empy vector and PBS group,they were respectively immunized by intramuscular injection in hind legs,The specific antibody of OrfV and cytokines IL-2,IL-4 and IFN-γ were detected by ELISA,and the proliferation of spleen lymphocytes was detected by MTT.The results showed that the ser-um antibody titer,IL-2 and IFN-γ cytokine levels of PVAX1-F1L+PVAX1-B2L combined immunization group were sign-ificantly higher than those of PVAX1-F1L and PVAX1-B2L group alone(P<0.05),There wa-s no significant difference in the level of IL-4,but it was significantly different from PVAX1 empt-y carrier and PBS group(P<0.01);the spleen lymphocyte proliferation level of PVAX1-F1L+PVAX1-B2L combined immunization group was significantly different from that of PVAX1-F1L a-nd PVAX1-B2L immunized groups(P<0.05),compared with PVAX1 empty vector and PBS gro-up.very significant(P<0.01).The results showed that PVAX1-F1L+PVAX1-B2L combined im-munization mice can enhance the secretion of OrfV-specific antibodies and cellular immune respo-nse,which provides a theoretical basis for the development of a new vaccine for Orf.