RHBV疫苗接种引起特异性细胞免疫反应.docVIP

RHBV疫苗接种引起特异性细胞免疫反应 　【摘要】 目的 研究小鼠免疫接种基因重组乙型肝炎表面抗原疫苗(recombinant HBV surface vaccine，rHBs)产生的特异性细胞免疫反应。方法 40只BALB/c小鼠随机分为0.65、1.25、2.5、5μg 4组，腹腔分别接种0.65、1.25、2.5、5μg的rHBs 1次或2次。初次免疫后4周或加强免疫后2周分离小鼠脾T淋巴细胞，分别进行脾T淋巴细胞释放IFNr、IL-2及细胞毒T淋巴细胞(cytotoxic T lymphocyte，CTL)实验；脾T淋巴细胞释放实验：用10g/ml rHBs刺激脾T淋巴细胞，3天后用ELISA方法检测培养液中IL-2及IFNr的浓度。CTL实验，脾T淋巴细胞用特异性多肽刺激，用51Cr释放实验检测CTL。结果 只接受单次免疫的0.65、1.25、2.5、5μg组小鼠脾T淋巴细胞释放的IL-2分别为5.48±8.88、9.28±6.98、28.53±14.32、64.69±20.88pg/ml，释放的IFNr分别为8.22±8.61、9.89±9.34、20.27±15.50、30.77±22.12pg/ml。接受加强免疫的0.65、1.25、2.5、5μg组小鼠脾T淋巴细胞释放的IL-2分别为5.75±5.04、102.53±67.52、177.13±91.12、332.10±124.31pg/ml，释放的IFNr分别为3.63±4.42、28.33±13.04、59.66±25.75、80.73±19.30pg/ml。CTL反应的靶细胞自然释放率为28.76%±13.45%；加强免疫组小鼠0.65、1.25、2.5、5组小鼠，CTL特异性释放率分别为(42.36±19.32)%、(91.21±22.97)%、(69.25±24.13)%、(51.49±21.661)%；只接受1次免疫接种的小鼠，其脾淋巴细胞特异性CTL特异性释放率分别为(32.27±15.35)%、(56.28±24.35)%、(44.34±18.65)%、(40.76±56)%。结论 小鼠接种rHBs后，脾T淋巴细胞产生特异性CTL反应，并特异性分泌IL-2、IFNr，反应强度与是否加强免疫及接种的剂量密切相关。 【关键词】 HBV疫苗；免疫接种；小鼠；脾T淋巴细胞；IL-2；IFNr；CTL反应 The cellular immune response to recombinant HBV vaccine 【Abstract】 Objective Hepatitis B virus(HBV) surface vaccine(rHBs) specific MHC class I-restricted cytotoxic T lymphocytes (CTL) responses induced by immunization with rHBs in murine were assayed.Methods BALB/c(H-2d) murine were injected intraperitoneal(i.p.)with rHBs at doses ranging from 0-6 μg and boosted after 2 weeks. 4 weeks after the last immunization，to measure the IL-2 and IFNr，spleen lymphocytes were harvested and restimulated with rHBs in experimental group or with PBS as control，4 weeks later after the last vaccination. For CTL assay，spleen cells were restimulated in vitro with specific peptides，which represents a Ld-restricted CTL epitope of the S-protein in H-2d BALB/c murine. The specific cytotoxic reactivity of in vivo primed and in vitro restimulated cells was tested against P815 cells impulsed with specific peptides and labeled with 51Cr. 4 hours release assay of 51Cr was performed. Results 2 weeks af