小鼠接种HBV疫苗引起的特异性细胞免疫反应

目的：研究小鼠免疫接种基因重组乙型肝炎表面抗原疫苗(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、5 μg 4组，腹腔分别接种0.65、1.25、2.5、5 μg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；分别进行以下实验：实验组用rHBs(10 mg/L)刺激脾T淋巴细胞，对照组用PBS代替rHBs刺激脾T淋巴细胞；3 d后用［3H］掺入法检测脾T淋巴细胞特异性增殖反应，以［3H］掺入的同位素counts·min-1值及刺激指数(SI, 实验组counts·min-1值/对照组counts·min-1值)表示。同时用ELISA方法检测培养液中白细胞介素-2(IL-2)及γ-干扰素(IFN-γ)的浓度。 结果：只接受单次免疫的0.65、1.25、2.5、5 μg组小鼠脾T淋巴细胞特异性增殖反应SI分别为1.55、1.93、2.41、2.811；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±20.88)ng/L，释放的IFN-γ分别为(8.22±8.61)、(9.89±9.34)、(20.27±15.50)、(30.77±22.12)ng/L。接受加强免疫的0.65、1.25、2.5、5 μg组小鼠脾T淋巴细胞特异性增殖反应SI分别为1.61、2.05、3.74、3.62；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±124.31)ng/L，释放的γ-干扰素分别为(3.63±4.42)、(28.33±13.04)、(59.66±25.75)、(80.73±19.30)ng/L。 结论： 小鼠接种rHBs后, 脾T淋巴细胞产生特异性增殖反应，并特异性分泌IL-2、γ-干扰素，反应强度与是否加强免疫及接种的剂量密切相关。

The specific T cell response in mice inoculated with HBV vaccine

AIM: To evaluate the specific cellular immune response in mice inoculated with the recombinant hepatitis B virus (HBV) surface vaccine (rHBs). METHODS: Spleen T lymphocyte reactivity to rHBs was assessed by a proliferation assay and cytokine secretion. BALB/c mouse were intraperitoneally inoculated with rHBs at doses of 0.65, 1.25, 2.5 or 5 μg for once or twice. 4 weeks later, spleen lymphocytes were harvested and restimulated with rHBs in experimental group or with PBS as control. The spleen lymphocytes were labeled with ［3H］-thymidine for 3 days. The ［3H］-thymidine incorporation was measured, which expressed as the incorporation of ［3H］ (counts·min-1) and stimulation index (SI) was calculated by the method of dividing the cpm obtained in the experimental group by that in control group. The content of IL-2 and IFN-γ secreted from the spleen lymphocyte were measured by the method of ELISA. RESULTS: 2 weeks after primary vaccination, the SI in 0.65, 1.25, 2.5 and 5 groups was 1.55, 1.93, 2.41, 2.81 ng/L, respectively. IL-2 was 5.48±8.88, 9.28±6.98, 28.53±14.32, 64.69±20.88 ng/L, respectively. IFN-γ was 8.22±8.61, 9.89±9.34, 20.27±15.50, 30.77±22.12 ng/L, respectively. 2 weeks after boost vaccination, the SI in 0.65, 1.25, 2.5 and 5 groups was 1.61, 2.05, 3.74, 3.62 ng/L, respectively. The IL-2 was 5.75±5.04, 102.53±67.52, 177.13±91.12, 332.10±124.31 ng/L, respectively. IFN-γ was 3.63±4.42, 28.33±13.04, 59.66±25.75, 80.73±19.30 ng/L, respectively. CONCLUSION: Specific proliferation activity and IL-2, IFN-γ secretion from the spleen lymphocytes of the mouse inoculated with rHBs are produced，that the strength is dependent on the dose of vaccination.