人乳头瘤病毒16型E5与IL-12联合基因疫苗的免疫活性

为了研制人乳头瘤病毒16型(HPV16)防治性疫苗,分析了HPV16 E5与IL-12联合基因疫苗的免疫活性。将构建的pcDNA3.1(+)/E5与pcDNA3.1(+)/IL-12联合免疫BALB/c小鼠,以ELISA测定小鼠血清中抗HPV16 E5 IgG水平、小鼠脾细胞培养上清中IFN-γ和IL-4含量;MTT法检测脾淋巴细胞增殖反应。结果显示末次免疫后,联合基因疫苗组和单基因疫苗组血清IgG A450值分别明显高于pcDNA3.1(+)组、pcDNA3.1(+)/IL-12组和PBS组(P<0.01);且联合基因疫苗组显著高于单基因疫苗组(P<0.01)。联合基因疫苗组和单基因疫苗组的IFN-γ和IL-4含量分别均明显高于pcDNA3.1(+)组、pcDNA3.1(+)/IL-12组和PBS组IFN-γ和IL-4含量(P<0.01),且联合基因疫苗组含量显著高于单基因疫苗组(P<0.01)。联合基因疫苗组和单基因疫苗组脾淋巴细胞刺激指数(SI)分别显著高于pcDNA3.1(+)组、pcDNA3.1(+)/IL-12组和PBS组(P<0.01);联合基因疫苗组与单基因疫苗组比较,SI差异无统计学意义(P>0.05)。结果表明HPV16 E5单基因疫苗以及与IL-12联合基因疫苗均能刺激机体产生较强的免疫应答,且联合基因疫苗优于单基因疫苗。

Immunological competence of combination DNA vaccine of human papillomavirus type 16 E5 and IL-12

To explore the prophylactic and treatmental vaccines for preventing the infections of human papillomavirus type 16(HPV16),the immunological competence of HPV16 E5 and IL-12 combination DNA vaccines was studied.The constructed plasmids pcDNA3.1(+) /E5 and pcDNA3.1(+) /IL-12 were inoculated into BALB/c mice.ELISA was used to detect anti-E5 IgG in sera of the mice,IFN-γ or IL-4 levels of the supernatant of mice spleen cells quantitatively.The proliferation response of the spleen cells was detected using MTT assay.After the last inoculation,A450 values of IgG from the combination(DNA vaccine) group pcDNA3.1(+)/E5+pcDNA3.1(+)/IL-12] or the single(DNA vaccine) grouppcDNA3.1(+)/E5] were significantly higher than that of IgG of group pcDNA3.1(+),group pcDNA3.1(+) /IL-12 or group PBS(P<0.01);meanwhile,the value from the combination group is obviously higher than that of the single group(P<0.01).The levels of IFN-γ or IL-4 from the combination group or the single group were significantly higher than those of group pcDNA3.1(+) group pcDNA3.1(+)/IL-12 or group PBS(P<0.01);there is the statistical difference between the combination group and the single group(P<0.01).The stimulation index(SI) of the spleen cells from the combination group or the single group is higher than that of group pcDNA3.1(+),group pcDNA3.1(+)/IL-12,and group PBS(P<0.01) respectively,but there is no significant difference between the combination group and the single group(P>0.05).It showed that HPV16 E5 DNA vaccine and combined DNA vaccine with IL-12 could induce strong specific immune response,and the combination DNA vaccine could make better immune effect.