The IFN-γ ELISpot method was established to detect the cellular immune level of a novel atten-uated 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-γ ELISPOT assay.The results show that the optimal stimulator of experimental hole is Ultraviolet inactivation HSV-2 particles(1× 108 CCID50/mL),the optimal cell culture time is 24-48 h,the optimal cell concentration is 3×106 cells/mL,the optimal immune route is subcutaneous injection.The IFN-γ ELISpot assay for the vaccine was es-tablished,which provides a method for detecting the cellular immune level of the vaccine.
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