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目的研究SA14-14-2株流行性乙型脑炎(乙脑)减毒活疫苗(Live Attenuated Japanese Encephalitis Vaccine,JEV-L)在小鼠体内的细胞免疫持久性。方法以SA14-14-2株JEV-L0.1ml皮下免疫BALB/c小鼠,分别于第14d和1、3、6、12个月处死,取脾脏,用酶联免疫斑点法(Enzyme-linked Immunospot,ELISPOT)检测脾淋巴细胞经JEV-L抗原刺激后分泌干扰素(Interferon,IFN)-γ和白细胞介素(Interleukin,IL)-2的水平,同时分离血清,混合后检测中和抗体水平。另一组相同免疫的小鼠于免疫后各时间点以P3毒株进行腹腔攻击,观察疫苗保护效果。结果 JEV-L免疫后14d细胞免疫水平达到高峰,持续1～3个月后开始略有下降,免疫后3～12个月的脾细胞IFN-γ斑点形成细胞(Spot Forming Cell,SFC)和IL-2SFC数量仍维持在较高水平。小鼠中和抗体水平在3个月内由1:20增加到1:80,12个月内仍维持在1:40。12个月内JEV-L免疫的小鼠均能很好地抵御乙脑强毒株病毒的腹腔攻击。结论 SA14-14-2株JEV-L能同时诱导细胞免疫反应和体液免疫反应,细胞免疫至少能维持12个月,JEV-L产生的保护效果也至少能持续12个月。
Objective To study the cellular immunity of SA14-14-2 live attenuated Japanese encephalitis vaccine (JEV-L) in mice. Methods BALB / c mice were immunized subcutaneously with SAE 14-14-2 strain JEV-L 0.1 ml and sacrificed on day 14 and 1, 3, 6 and 12 respectively. The spleens were harvested and stained with enzyme-linked immunosorbent assay Immunospot and ELISPOT) were used to detect the levels of Interferon (IFN) -γ and Interleukin (IL) -2 secreted by spleen lymphocytes stimulated by JEV-L antigen. At the same time, the serum was separated and the levels of neutralizing antibodies . Another group of mice immunized with the same immunization at each time point after the IP strain of intraperitoneal attack to observe the vaccine protective effect. Results The level of cellular immunity peaked at 14 days after JEV-L immunization, and began to decline slightly after 1-3 months. After immunized for 3 to 12 months, IFN-γ spot-forming cells (SFC) and IL -2SFC quantity is still maintained at a high level. Mice neutralizing antibody levels increased from 1:20 to 1:80 in 3 months, and JEV-L immunized mice remained within 1: 40.12 months after 12 months Severe strains of the virus intraperitoneal attack. Conclusion The JEV-L strain SA14-14-2 induces both cellular and humoral immune responses. The cellular immunity can be maintained for at least 12 months. The protective effect of JEV-L can last for at least 12 months.