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目的 构建HBV囊膜中蛋白核酸疫苗表达载体并免疫小鼠 ,观察白细胞介素 18(IL 18)对基因免疫的辅助作用。方法 构建质粒 pVR10 12 M、pcDNA 3 .1- IL 18,肌内注射法免疫 2 5只Balb/c小鼠 ,3组小鼠分别注射 10 0 μgpVR10 12 ,pVR10 12 M ,pVR10 12 M和 pcDNA 3 .1- IL 18质粒 ,每 2周 1次 ,共 3次。每次免疫 2周后眼眶采血 ,检测血清抗 HBs,第 3次免疫后 2周应用乳酸脱氢酶检测法验证特异性细胞杀伤率。结果 经注射上述质粒后 ,可观察到注射 pVR10 12 M组的小鼠随免疫次数的增加 ,抗 HBs阳性率、抗体滴度均逐步增高 ;同时联用质粒 pcDNA 3 .1- IL 18的小鼠抗 HBs阳性率、抗体滴度均较单独应用 pVR10 12 M组为低 (第 3次免疫后抗 HBs阳性率、抗体滴度两组比较P <0 .0 5 )。细胞毒性实验证实联用组的细胞杀伤率为 (89.0 2± 15 .5 4) % ,较单独应用pVR10 12 M组 (83 .0 8± 14 .0 2 ) %为高 ,但两组之间差异无显著性。 结论 注射质粒 pVR10 12 M可诱导小鼠产生足量的抗 HBs ,并可检测出特异性细胞免疫反应 ;联合应用IL 18对特异性体液免疫有抑制作用 ,对特异性细胞免疫可能有增强作用
Objective To construct HBV DNA vaccine expression vector and immunize mice to observe the adjuvant effect of interleukin 18 (IL 18) on gene immunity. Methods Plasmids pVR10 12 M and pcDNA 3 .1-IL 18 were constructed and 25 Balb / c mice were immunized intramuscularly. Three groups of mice were injected with 10 μg of pVR10 12, pVR10 12 M, pVR10 12 M and pcDNA3 .1-IL 18 plasmid once every 2 weeks for 3 times. Serum anti-HBs was collected 2 weeks after each immunization, and specific cytotoxicity was tested by lactate dehydrogenase assay 2 weeks after the third immunization. Results The mice injected with pVR10 12 M were observed to have an increased frequency of immunization and the positive rate of anti-HBs and antibody titers were increased after injection of the above plasmids. In addition, the mice immunized with pcDNA3.1-IL18 The positive rate of anti-HBs and antibody titers were lower than that of pVR10 12 M alone (the positive rate of anti-HBs after the third immunization, the antibody titer of the two groups were P <0.05). Cytotoxicity test showed that the cell killing rate of the combination group was (89.0 ± 2.54)%, which was higher than that of the pVR10 12 M group (83.0 ± 14.0 ± 2)% group alone No significant difference. Conclusion Injection of plasmid pVR10 12 M can induce a sufficient amount of anti-HBs in mice and detect specific cellular immune responses. Combined use of IL 18 can inhibit specific humoral immunity and enhance specific cellular immunity