地方株HPV16E7基因疫苗诱发的小鼠免疫反应

目的评价地方株HPV16E7基因疫苗诱导的免疫反应。方法从先期构建的地方株HPV16E7基因重组质粒pGEM-T-E7经限制性核酸内切酶酶切,获得E7基因亚克隆到真核表达载体pcDNA3.1(-)中,构建地方株pcDNA-E7基因疫苗,转染CHO细胞,通过IFA试验验证E7蛋白的表达。用pcDNA-E7肌注方法免疫6周龄BALB/c小鼠,同时设pcDNA空载体为阴性对照,于第1天、第15天和第43天共免疫3次,用IFA和MTT法分析基因免疫在小鼠体内诱导的免疫应答。结果pcDNA-E7免疫小鼠后能检测出特异的抗体,而原载体免疫后不能产生特异抗体;但2组刺激淋巴细胞增殖的能力差异不显著。结论地方株HPV16E7基因疫苗可诱导产生小鼠免疫反应。

Immune responses induced by DNA vaccine for local strain of human papillomavirus E7 in mice

Objective:To evaluate the ability of DNA vaccine of local strain of HPV (human papilloma virus)16E7 in inducing specific humoral immune responses.Methods:The recombinant plasmid pGEM-T-E7 of HPV16E7 local strain was enzyme-digested with restriction endonucleases to get E7 gene subclone. E7 was subcloned into the eukaryotic expression vector pcDNA3.1(-) . The CHO cells were transfected with pcDNA-E7. E7 protein expression was identified by immunofluorescence assay. BALB/c mice (6-week-old) was immunized with pcDNA-E7 3 times intramuscularly at d 1,d 15 and d 43,and pcDNA vector was as the control. The immune response was measured by indirect immunofluorescence assay (IFA) and lymphocyte proliferation was determined by MTT method. Results: Specific anti-HPV16E7 antibodies could be detected in pcDNA-E7 immunized mice but not in pcDNA-immunized mice. The difference in lymphocyte proliferation was not statistically significant between pcDNA-E7 immunized mice and the control group. Conclusions:DNA vaccine of HPV16E7 local strain could induce immune responses in mice.