新型减毒HSV-2疫苗IFN-γELISpot检测方法的建立

Establishment of IFN-γELISpot Assay for a New Attenuated HSV-2 Vaccine

通过比较不同刺激物、细胞培养时间、细胞接种浓度、免疫途径,建立检测新型减毒HSV-2疫苗细胞免疫水平的IFN-γELISpot方法。对C57BL/6小鼠进行减毒HSV-2疫苗免疫,第14天加强免疫,第21天分离小鼠脾脏淋巴细胞,采用IFN-γELISpot检测该疫苗的细胞免疫水平。结果表明:最佳实验孔刺激物为紫外灭活30 min、原滴度为10^(8)CCID_(50)/mL的疫苗全病毒颗粒,最佳细胞培养时间为24~48 h,最佳细胞接种浓度为3×10^(6)cells/mL,最佳免疫途径是皮下注射。实验建立了针对该疫苗的IFN-γELISpot方法,为该疫苗提供了一种检测细胞免疫水平的方法。

The IFN-γELISpot method was established to detect the cellular immune level of a novel attenuated HSV-2 vaccine by comparing the experimental conditions such as different stimulators,cell culture time,cell inoculation concentration,and immune pathway.C57BL/6 mice were immunized with the novel attenuated HSV-2 vaccine,and strengthened on the 14th day.Spleen lymphocytes were isolated on the 21st day.The cellular immune level of the vaccine was detected by IFN-γELISPOT assay.The results show that the optimal stimulator of experimental hole is Ultraviolet inactivation HSV-2 particles(1×10^(8)CCID_(50)/mL),the optimal cell culture time is 24-48 h,the optimal cell concentration is 3×10^(6)cells/mL,the optimal immune route is subcutaneous injection.The IFN-γELISpot assay for the vaccine was established,which provides a method for detecting the cellular immune level of the vaccine.