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目的研究人乳头瘤病毒18型E2基因(HPV18-E2)疫苗的免疫效果及白细胞介素-12(IL-12)对其免疫作用的影响。方法选6~8周龄Balb/c雌性小鼠35只,随机分为5组,即PBS组、pcDNA3.1(+)空质粒组、pcD-NA3.1(+)/E2组、pcDNA3.1(+)/IL-12组和pcDNA3.1(+)/E2+pcDNA3.1(+)/IL-12组,每组7只。小鼠肌肉注射DNA疫苗,200μg/次。隔2周免疫1次,共免疫4次。于0、2、4、6周剪尾取血,第8周摘眼球放血。ELISA测血清中的特异性IgG抗体及小鼠脾淋巴细胞培养上清IFN-γ、IL-4,MTT比色法检测淋巴细胞的增殖。结果免疫8周pcDNA3.1(+)/E2及pcDNA3.1(+)/E2+pcDNA3.1(+)/IL-12疫苗组抗体IgG A值显著升高,与对照组比较差异有统计学意义(P<0.01),pcDNA3.1(+)/E2及pcDNA3.1(+)/E2+pcDNA3.1(+)/IL-12组小鼠脾淋巴细胞培养上清IFN-γ、IL-4水平明显高于其他对照组,差异有统计学意义(P<0.01),pcDNA3.1(+)/E2+pcDNA3.1(+)/IL-12组小鼠脾淋巴细胞增殖活性明显增强,与pcDNA3.1(+)/E2组比较,差异无统计学意义(P>0.05),与其他对照组比较差异有统计学意义(P<0.01)。结论pcDNA3.1(+)/E2+pcDNA3.1(+)/IL-12核酸疫苗联合免疫小鼠能够有效的诱导细胞免疫和体液免疫应答,且免疫效应比pcDNA3.1(+)/E2基因疫苗强。
Objective To investigate the effect of HPV18-E2 vaccine and the effect of interleukin-12 (IL-12) on its immunological function. Methods 35 female Balb / c mice aged 6-8 weeks were randomly divided into 5 groups: PBS group, pcDNA3.1 (+) empty plasmid group, pcD-NA3.1 (+) / E2 group and pcDNA3. 1 (+) / IL-12 group and pcDNA3.1 (+) / E2 + pcDNA3.1 (+) / IL-12 group. Mice were injected intramuscularly with DNA vaccine, 200 μg / dose. Immunization every 2 weeks, a total of 4 immunizations. Tail blood was collected at 0, 2, 4 and 6 weeks, and blood was taken at 8 weeks. Specific IgG antibodies in ELISA serum and IFN-γ, IL-4 and MTT colorimetric assay were used to detect the proliferation of lymphocytes. Results The IgG A value of the antibody immunized with pcDNA3.1 (+) / E2 and pcDNA3.1 (+) / E2 + pcDNA3.1 (+) / IL-12 vaccine was significantly increased at 8 weeks after immunization (P <0.01). The levels of IFN-γ and IL-6 in the splenic lymphocytes of pcDNA3.1 (+) / E2 and pcDNA3.1 (+) / E2 + pcDNA3.1 (+) / 4 levels were significantly higher than those in other control groups (P <0.01). The proliferation activity of splenic lymphocytes in mice immunized with pcDNA3.1 (+) / E2 + pcDNA3.1 (+) / IL- Compared with pcDNA3.1 (+) / E2 group, the difference was not statistically significant (P> 0.05), and there was significant difference compared with other control groups (P <0.01). Conclusion The combination of pcDNA3.1 (+) / E2 + pcDNA3.1 (+) / IL-12 nucleic acid vaccine can effectively induce cellular and humoral immune responses, and the immune effect is better than that of pcDNA3.1 (+) / E2 Strong vaccine.