重组人IL-12真核表达载体的基因佐剂功能

目的探讨重组人IL-12真核表达载体(pcDNA6-p70)对核酸疫苗(HCMV pp65基因重组腺病毒,Adeno-pp65)的基因佐剂功能。方法pcDNA6-p70与Adeno-pp65共免疫BALB/c小鼠,每2周1次,共4次,同时设立Adeno-pp65对照组及生理盐水对照组。于初次免疫后第6周末尾静脉取血,第9周处死小鼠,分离脾细胞,分别检测细胞内细胞因子水平、CTL杀伤活性、特异性淋巴细胞增生和IFN-γ水平,并观察pcDNA6-p70对小鼠的安全性。结果共免疫组HCMV特异性CD4/IFN-γ、CD8/IFN-γ双标记阳性细胞的百分率均高于Adeno-pp65组及生理盐水对照组,差异均有显著意义;其CTL杀伤活性、HCMV特异性淋巴细胞增生水平及脾淋巴细胞特异性IFN-γ释放水平均高于Adeno-pp65组和生理盐水对照组,差异均有显著意义。pcDNA6-p70肌肉注射昆明小鼠,其体温、体重及一般情况与对照组差异均无显著意义。结论pcDNA6-p70与Adeno-pp65共免疫可提高核酸疫苗的免疫效果,且具有较好的安全性。

Genetic Adjuvanticity of A Eukaryotic Expression Vector for Recombinant Human IL-12

Objective To explore the genetic adjuvanticity of eukaryotic expression vector pcDNA6-p70 for recombinant human IL-12 (rhIL-12) in immunization with DNA vaccine Adeno-pp65 (recombinant adenovirus with HCMV pp65 gene). Methods Eighteen BALB / c mice were divided into one test and two control groups, 6 for each. The mice in test group were co-immunized with pcDNA6-p70 and Adeno-pp65 every 2 weeks for 4 times, and those in two control groups with Adeno-pp65 and physiological saline respectively. Collect the sera of mice 6 weeks after the first immunization for determination of IFN-γ level. Kill the mice 9 weeks after the first immunization and isolate splenocytes for determination of cellular immunity, CTL killing activity, IFN-γ level and specific lymphocyte proliferation. Meanwhile, the safety of pcDNA6-p70 was observed in KM mice. Results The percentages of splenocytes positive for CD4 / IFN-γ and for CD8 / IFN-γ as well as CTL killing activity, the proliferation level of HCMV specific lymphocytes and the release level of splenic lymphocyte specific IFN-γ of mice in test group were significantly higher than those in the two control groups. However, the body temperature, bodyweight and general status of KM mice inject i. m. with pcDNA6-p70 showed no significant difference with those of control. Conclusion Eukaryotic expression vector pcDNA6-p70 enhanced the immune effect of Adeno-pp65 and showed good safety.