乙型肝炎疫苗加强免疫诱导小鼠细胞免疫应答相关因素的研究

Study of the Influences Related on Cellular Immune Response in Mice after Hepatitis B Vaccine Revaccination

目的探讨现有乙型肝炎(乙肝)疫苗(Hepatitis B Vaccine,HepB)在小鼠(H-2d)加强免疫中,不同免疫剂量、免疫时间及检测时间对细胞免疫检测的影响。方法采用三种疫苗剂量,两种加强免疫程序免疫BALB/c小鼠,即初次免疫后14d加强免疫和初次免疫后28d加强免疫,分别于加强免疫后7d、14d分离小鼠脾单个核细胞(Mononuclear Cell,MNC),使用酶联免疫斑点实验(Enzyme-linked Immunospot Assay.ELISPOT)测定小鼠MNC的干扰素(Interferon,IFN)-γ、白细胞介素(Interleukin,IL)-2和IL-5斑点形成细胞数(Spots-forming Cell,SFC)。结果加强免疫后7d,IFN-γ和IL-2的分泌均处于较高水平,而加强免疫后14d检测时两者SFC均出现下降;同时14d时可见ELISPOT阳转率也下降,而加强免疫后7d时阳转率均为100%;14d加强免疫与28d加强免疫程序相比,前者在加强免疫后7d、14d IL-2分泌水平均高于后者,在28d加强免疫后7d检测IL-5阳转率均较高,14d时其阳转率出现下降。结论 HepB在第14d进行加强免疫能够诱导较高的细胞免疫反应,加强免疫后7d可检测到较高的细胞免疫反应。

Objective To investigate the effect of cellular immunity detection after hepatitis B vaccine immunization in mice with different booster immunization procedure,doses,immunization time and detection time.Methods Three vaccine doses and two types of booster immunization procedure were applied.Booster immunization was administered 14 or 28 days after prime immunization on BALB/c mice,and 7 days,14 days after booster immunization enzyme-linked Immunospot（ELISPOT）assay were conducted to evaluate interferon（IFN）-γ,interleukin（IL）-2 and IL-5 spot-forming cells（SFC）of splenic mononuclear cells（MNC）.Results 7days after boost vaccination,IL-2 and IFN-γ demonstrated a high level and positive rate,while both of them became decreased when detected 14 days after vaccination.Comparing the two booster procedures,the former can be detected high IL-2 secreting level in both detection time point,while in the latter procedure the positive rate of IL-5 showed an increase according to the vaccine dose.Conclusion Hepatitis B vaccine can induce higher cellular immune response when boosted 14 days after prime immunization.At the 7th day after boost vaccination,cellular immune response was higher than the other detection time point.