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目的探讨CpG ODN1826为佐剂的布氏菌DNA疫苗对C57/BL6小鼠的免疫效果。方法将C57/BL6小鼠随机分为3组:重组质粒(pc DNA3.1-L7/L12)+CpG ODN1826组(Vaccine+CpG组,每只注射20μg重组质粒和10μg CpG ODN1826)、重组质粒组(Vaccine组,每只注射20μg重组质粒)和对照组(注射生理盐水),注射部位均为后腿胫骨前肌。各组均于初次免疫后2周加强免疫1次。于加强免疫后2周,采用ELISA法检测各组小鼠血清中细胞因子水平;MTT掺入法检测小鼠脾细胞非特异性增殖效应;HE染色观察小鼠脾脏组织学变化。结果与Vaccine组和对照组相比,Vaccine+CpG组小鼠血清中Th1型细胞因子IL-2、IL-12、IFNγ含量均显著升高(P均<0.01);脾细胞增殖效应明显增强(P<0.05);脾脏生发中心明显变大,细胞增殖更活跃。结论 CpG ODN1826为佐剂的布氏菌DNA疫苗能够明显诱导C57/BL6小鼠产生Th1型为主的免疫应答,增强脾脏淋巴细胞的增殖反应。
Objective To investigate the immunogenicity of C57 / BL6 mice induced by DNA vaccine with CpG ODN1826 adjuvant. Methods C57 / BL6 mice were randomly divided into 3 groups: pcDNA3.1-L7 / L12 + CpG ODN1826 group (Vaccine + CpG group, 20μg recombinant plasmid and 10μg CpG ODN1826), recombinant plasmid group (Vaccine group, each injection of 20μg recombinant plasmids) and control group (injection of saline), the injection site are hind tibialis anterior muscle. Each group was boosted once every 2 weeks after the first immunization. Two weeks after booster immunization, the levels of cytokines in serum of mice in each group were detected by ELISA. The non-specific proliferation of spleen cells was detected by MTT assay. The histological changes of spleen in mice were observed by HE staining. Results Compared with Vaccine group and control group, the levels of Th1 cytokines IL-2, IL-12 and IFNγ in Vaccine + CpG group were significantly increased (all P <0.01), and the proliferation of spleen cells was significantly increased P <0.05); spleen germinal center was significantly larger, more active cell proliferation. Conclusion The DNA vaccine against B. bovis induced by CpG ODN1826 can significantly induce Th1-type immune response in C57 / BL6 mice and enhance the proliferation of spleen lymphocytes.