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目的观察SARS S DNA疫苗经不同的免疫途径及免疫次数,所诱导的免疫效果之间的差异。方法 C57BL/6小鼠经肌肉和皮下途径初次或加强免疫SARS S DNA疫苗,取其脾和肺淋巴细胞经S抗原多肽刺激以后,应用ELISA、ELISPOT和FACS等检测诱导细胞产生细胞因子情况。结果肌肉初次免疫后产生IFN-γ的量较低(均值脾:2.7 pg/ml;肺:68.9 pg/ml),加强免疫2次或3次后IFN-γ水平明显提高(脾:809.2～1398.7 pg/ml;肺:202.3～534.6 pg/ml),与初次免疫比较差异有统计学意义(脾P<0.001,肺P<0.05)。皮下二次免疫后IFN-γ产生量为54.3～106.1 pg/ml,但三次免疫后IFN-γ量显著提高均值达到729.1 pg/ml与对照组比较P<0.05。二次免疫后抗原特异性阳性细胞的频率肌肉免疫(188个/2×105细胞)高于皮下免疫(57个/2×105细胞),两者差异有统计学意义(P<0.05),而免疫3次后2种免疫途径可诱导数量相当的特异性阳性细胞(169～198个/2×105细胞,P>0.05)。S抗原特异性CD4+T细胞亚群以分泌IL-2为主,而抗原特异性CD8+T细胞分泌IFN-γ为主。结论经肌肉和皮下2种途径注射SARS S DNA疫苗免疫后,均可诱导小鼠产生特异性细胞免疫应答;加强免疫后诱导免疫应答明显强于初次免疫;肌肉免疫两次或皮下免疫三次后可达最佳免疫效果。
Objective To observe the difference between immune effects induced by SARS S DNA vaccine through different routes of immunization and frequency of immunization. Methods C57BL / 6 mice were immunized with SARS S DNA vaccine by primary and secondary immunization with subcutaneous or subcutaneous route. The spleen and lung lymphocytes were stimulated with S antigen peptide. The cytokines were induced by ELISA, ELISPOT and FACS. RESULTS: The amount of IFN-γ produced after the initial immunization of muscle was lower (mean spleen: 2.7 pg / ml; lung: 68.9 pg / ml), and the level of IFN-γ significantly increased after 2 or 3 times of intensive immunization (spleen: 809.2-1398.7 pg / ml; lung: 202.3-536.6 pg / ml) compared with the primary immunization. There was significant difference (P <0.001, P <0.05). The amount of IFN-γ produced after subcutaneous immunization was 54.3-106.1 pg / ml, but the IFN-γ level increased significantly after the three immunizations reached 729.1 pg / ml compared with the control group (P <0.05). The frequency of antigen-specific positive cells after secondary immunization was higher than that of subcutaneous immunized mice (188 cells / 2 × 105 cells) (57 cells / 2 × 105 cells), and the difference was statistically significant (P <0.05) After 3 immunizations, the two immune pathways induced a considerable number of specific positive cells (169-198 cells / 2 × 105 cells, P> 0.05). S antigen-specific CD4 + T cell subsets secreted IL-2, while antigen-specific CD8 + T cells secreted IFN-γ. Conclusions Both SARS S DNA vaccine injected intramuscularly and subcutaneously could induce specific cellular immune response in mice. The immune response induced by SARS S was stronger than that of primary immunization, and twice after immunization with muscle or subcutaneously The best immune effect.