【摘要】 目的探索并建立带状疱疹减毒活疫苗免疫大鼠脾细胞的体外IFN-γ释放检测方法 ,并建立评价带状疱疹疫苗免疫应答的检测平台。方法取雌性SD大鼠免疫后4、6、8、12周的血清及脾细胞,分别用gp-ELISA法定量检测血清IgG水平,用双抗体夹心ELISA法定量检测脾细胞体外IFN-γ释放水平,用SPSS 16.0软件分析检测数据。结果大鼠IFN-γ释放受到脾细胞数、培养时间、刺激物种类的影响,脾细胞数为1×107个/ml、用带状疱疹疫苗刺激6 d的IFN-γ释放量[(6 617±1.33)pg/ml]达到峰值,协同刺激因子可加速脾细胞体外IFN-γ释放速度,刺激2 d后IFN-γ释放量[(6 016±1.46)pg/ml]达到峰值,与疫苗单独刺激组6 d的峰值水平一致。大鼠免疫后血清IgG水平与脾细胞释放的IFN-γ水平随带状疱疹疫苗免疫次数增加的变化趋势一致,皆于第2次免疫后达到峰值期。结论通过对影响带状疱疹疫苗免疫大鼠脾细胞体外IFN-γ释放因素的研究,建立了大鼠脾细胞体外IFN-γ释放检测平台,且发现协同刺激因子可加速而不额外增加脾细胞体外IFN-γ释放量,为快速检测大鼠脾细胞体外IFN-γ释放量提供了方法;同时验证了gp-ELISA检测的IgG与双抗体夹心检测的IFN-γ具有一致性。更多还原

【Abstract】 Exploring VZV-specific IFN-γ release in vitro in order to establish an assay for detecting the level of immune response induced by herpes zoster vaccine.Immune sera and spleen lymphocytes were taken from female SD rats at weeks 4,6,8,and 12 post-immunization for evaluating IgG level and IFN-γ release by a quantitative gp-ELISA and a quantitative ELISA,respectively.All data were analyzed by SPSS 16.0 statistical software.The results showed that the appropriate IFN-γ release depends on the density of spleen cells,culture time and stimulus types.PMA,Ion,IL-2 could simultaneously increase the speed of IFN-γ release compared with vaccine alone.And the maximum release of IFN-γ is about 6 000-7 000 pg/ml.With the increase of the immunization times,IgG level and IFN-γ release reached a plateau after the second booster and followed a consistent tendency.In conclusion,we established an assay for detecting the level of immune response induced by herpes zoster vaccine successfully.And costimulatory molecules could significantly accelerate IFN-γ release in vitro.Both IgG level and IFN-γ release stimulated either vaccine alone or costimulatory molecules have a consistent tendency.更多还原