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目的评价IgG Fc段编码基因不同构建方式对乙型脑炎(JE)DNA疫苗细胞免疫应答的影响。方法以套式RT-PCR法获取BALB/c鼠IgG Fc编码基因,构建含乙型脑炎病毒(JEV)pr ME蛋白与IgG Fc编码基因以及单纯IgG Fc编码基因重组质粒,分别命名pJME/IgG Fc和pIgG Fc,以不同免疫原肌注免疫BALB/c小鼠,经流式细胞仪检测不同免疫原免疫鼠后脾T淋巴细胞亚群及Th细胞内细胞因子(IFN-γ、IL-4)变化,乳酸脱氢酶(Lactate dehydrogenase,LDH)法测定细胞毒性T淋巴细胞(Cytotoxic T lymphocytes,CTL)活性。数据进行单因素方差分析。结果 pJME/IgG Fc组CD4+T淋巴细胞比例为(66.64±5.84)%,明显高于pJME+pIgG Fc、pJME、JE灭活疫苗和pc DNA3.1(+)组(P<0.05),pJME+pIgG Fc组CD4+T淋巴细胞比例(60.55±2.09)%,高于pJME组(52.86±1.92)%,也高于JE灭活疫苗组(55.77±1.62)%(P<0.05),pc DNA3.1(+)组CD4+T淋巴细胞比例为(37.82±4.93)%,明显低于其他组(P<0.05);pJME/IgG Fc与pJME+pIgG Fc免疫组CD8+T细胞比例较空载体pc DNA3.1(+)及灭活疫苗组升高(P<0.05)。pJME/IgG Fc免疫组CTL活性为(46.92±1.97)%,明显高于其它组(P<0.05)。pJME/IgG Fc、pJME+pIgG Fc免疫组IFN-γ+CD4+T淋巴细胞比例分别为(37.90±4.79)%、(21.53±4.61)%,明显高于其它组(P<0.05)。结论相对于pJME+pIgG Fc联合免疫组,pJME/IgG Fc融合质粒免疫组能够诱导更强的细胞免疫反应。
Objective To evaluate the effect of different coding methods of IgG Fc fragment on the cellular immune response of Japanese encephalitis (JE) DNA vaccine. Methods The coding sequence of IgG Fc of BALB / c mice was obtained by nested RT-PCR, and the recombinant plasmids containing Japanese encephalitis virus (JEV) pr ME protein, IgG Fc gene and IgG Fc-encoding gene were constructed and named as pJME / IgG Fc and pIgG Fc were injected intraperitoneally to BALB / c mice with different immunogens. The splenic T lymphocyte subsets and Th cell cytokines (IFN-γ, IL-4 ), And the activity of cytotoxic T lymphocytes (CTL) was measured by Lactate dehydrogenase (LDH). Data were subjected to one-way analysis of variance. Results The percentage of CD4 + T lymphocytes in pJME / IgG Fc group was (66.64 ± 5.84)%, which was significantly higher than that in pJME + pIgG Fc, pJME, JE inactivated vaccine and pcDNA3.1 (+) group (60.55 ± 2.09)% in the pIgG Fc group, which was higher than that in the pJME group (52.86 ± 1.92)% and also higher than that in the JE inactivated vaccine group (55.77 ± 1.62)% (P <0.05) (37.82 ± 4.93)%, respectively (P <0.05). The proportion of CD8 + T cells in pJME / IgG Fc group and pJME + pIgG Fc group was significantly lower than that in other groups pcDNA3.1 (+) and inactivated vaccine group increased (P <0.05). The CTL activity of pJME / IgG Fc immunized group was (46.92 ± 1.97)%, which was significantly higher than other groups (P <0.05). The ratios of IFN-γ + CD4 + T lymphocytes in pJME / IgG Fc and pJME + pIgG Fc groups were (37.90 ± 4.79)% and (21.53 ± 4.61)%, respectively, which were significantly higher than those in the other groups (P <0.05). Conclusion Compared with the pJME + pIgG Fc combined immunization group, the pJME / IgG Fc fusion plasmid immunized group can induce a stronger cellular immune response.