【摘要】 目的小鼠经皮下SARS-CoVS DNA疫苗免疫后,研究其特异性T细胞及相关细胞因子的特性。方法SARS-CoVS DNA疫苗免疫BALBPc小鼠后,获取淋巴细胞悬液。经S抗原多肽刺激后,采用ELISA检测细胞培养上清液中IFN-γ的水平,利用流式细胞仪在单个细胞水平上检测IFN-γ和IL-2的表达及其关系。结果当S混合多肽刺激后,DNA疫苗免疫小鼠的淋巴细胞产生大量的IFN-γ,与对照鼠相比差异有统计学意义(P<0.01)。细胞亚群分析的结果表明,IFN-γ+和IL-2+的CD4+T细胞百分率明显高于CD8+T细胞。单独产生IL-2的细胞占大多数,其次为IFN-γ和IL22双阳性细胞,只产生IFN-γ的细胞很少。结论SARS-CoVS DNA疫苗免疫小鼠后可以诱导抗原特异性CD4+和CD8+T细胞的产生。更多还原

【Abstract】 Objective To elucidate the characterization of antigen-specific T cells from mice after immunization with SARS-CoV S DNA vaccine. Methods BALB/c mice were immunized subcutaneously with SARS-CoV S DNA vaccine. By day 7 after vaccination, single cells from lymph nodes, spleen and lung were prepared and incubated with pooled SARS-S peptides. The level of IFN-γ in the culture supernatants was assayed by ELISA. The frequency of IFN-γ and IL-2-producing cells were determined at a single cell level with FACS. Results After stimulation with SARS-CoV S peptides, significantly higher level of IFN-γ in culture supernatants from spleen and lung were observed in the group of SARS-CoV S DNA immunized mice than in those of control ones(P<0.01). Further analysis indicated that most of cells expressed IL-2 alone, some cells produced both IL-2 and IFN-γ, and very few cells were IFN-γ~+ alone. In addition, higher percentage of IFN-γ~+ and IL-2~+ T cells were detected in lymph nodes and spleens than in lungs. ZConclusion Vaccination of mice with SARS-CoV S DNA induces in the generation of antigen-specific CD4~+/CD8~+ T cells, which may plays an important role in the control and prevention of SARS infection.更多还原