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目的探讨不同种类乙型肝炎(乙肝)疫苗免疫小鼠后诱导抗原特异性细胞因子反应的差异及单个核细胞产生TH1型细胞因子的能力,评价不同种类乙肝疫苗的免疫原性。方法应用重组汉逊酵母、酿酒酵母、CHO细胞来源的乙肝疫苗分别免疫BALB/c小鼠(H-2d),于免疫后4、7、10、14、25和35 d处死,分离脾单个核细胞(Mononuclear cell,MNC),经HBsAg体外刺激后,采用液相芯片酶联(Luminex)技术测定小鼠IFNγ、IL-2和TNF-α分泌水平。结果 3种乙肝疫苗诱导小鼠产生IFNγ和TNF-α的峰值均出现在免疫后10 d,之后开始下降;IL-2水平呈逐渐上升趋势。免疫后10和14 d,酿酒酵母疫苗组诱导小鼠产生IFNγ的水平显著高于CHO疫苗组(P<0.01);免疫后10、25和35 d,汉逊酵母疫苗组诱导小鼠产生IL-2的水平显著高于CHO疫苗组(P<0.05);免疫后10、14和35 d,酿酒酵母疫苗组诱导小鼠产生TNF-α的水平显著高于CHO疫苗组(P<0.05);免疫后7和25 d,汉逊酵母疫苗组诱导小鼠产生TNF-α的水平显著高于CHO疫苗组(P<0.05)。结论 不同种类乙肝疫苗诱导小鼠的细胞免疫反应强度与变化趋势不同。酵母疫苗诱导的TH1型细胞因子水平高于CHO疫苗。
Objective To investigate the difference of antigen-specific cytokine response induced by different kinds of Hepatitis B vaccine and the ability of mononuclear cells to produce TH1-type cytokines and to evaluate the immunogenicity of different kinds of Hepatitis B vaccines. Methods BALB / c mice (H-2d) were immunized with recombinant Hepatitis B, Saccharomyces cerevisiae and CHO cell-derived Hepatitis B vaccine and were sacrificed at 4, 7, 10, 14, 25 and 35 d after immunization, (MNC) cells were stimulated with HBsAg in vitro. The levels of IFNγ, IL-2 and TNF-α in mice were measured by Luminex. Results The peak values of IFNγ and TNF-α produced by three kinds of hepatitis B vaccines were all appeared on the 10th day after immunization, and then decreased. The level of IL-2 showed a gradual upward trend. The levels of IFNγ induced by Saccharomyces cerevisiae vaccine group was significantly higher than that of CHO vaccine group at 10 and 14 days after immunization (P <0.01). At 10, 25 and 35 days after immunization, the Hansenula vaccine group induced IL- 2 was significantly higher than that of CHO vaccine group (P <0.05). At 10, 14 and 35 days after vaccination, the level of TNF-α induced by Saccharomyces cerevisiae vaccine group was significantly higher than that of CHO vaccine group (P <0.05) After 7 and 25 d, the level of TNF-α production in Hansenula vaccine group was significantly higher than that in CHO vaccine group (P <0.05). CONCLUSION: The intensity and changing tendency of cellular immune response in different kinds of hepatitis B vaccine are different. Yeast vaccines induce higher levels of TH1-type cytokines than CHO vaccines.