小鼠接种HBV疫苗引起的特异性细胞免疫反应

The specific T cell response in mice inoculated with HBV vaccine

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、5μg4组,腹腔分别接种0·65、1·25、2·5、5μg的rHBs1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(10mg/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;3d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、5μg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、5μg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。

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 was5.48 ± 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- γsecretion from the spleen lymphocytes of the mouse inoculated with rHBs are produced, that the strength is dependent on the dose of vaccination.