CAJ | 학술논문

目的评价HPV16 E6E7的复制缺陷型重组5型腺病毒(PK-HPV-ad5)治疗性疫苗对实验小鼠免疫应答和抗肿瘤的生物学效应.方法使用基因重组技术构建PK-HPV-ad5疫苗,并通过小鼠免疫试验,检测小鼠总抗体和特异性IFNγ,同时将造模小鼠分成疫苗组和对照组,分别对其进行抑瘤试验、TC-1肿瘤细胞挑战试验和肿瘤切除后防复发试验.结果 HPV16 E6E7诱导的总抗体第12天的水平相对较高(1:400～1:600);3批次疫苗特异性IFNγ在第14天与对照组比较分别升高8.6、5.9和8.9倍,差异有统计学意义(t=15.721、6.967和14.342,P均＜0.01).抑瘤试验表明疫苗剂量为107IU/只时小鼠肿瘤生长率为0,与对照组比较差异有统计学意义(确切概率法,P＜0.01),3批次疫苗验证有效剂量为107IU/只时肿瘤抑制率可达80％(8/10)以上.TC-1肿瘤细胞挑战试验结果显示:小鼠先接种疫苗能引起特异性的免疫应答,并能保护90％(9/10)的小鼠免受TC-1肿瘤细胞的攻击;肿瘤切除后防止复发试验提示在注射相同剂量疫苗时,对104个/只和105个/只肿瘤细胞造模小鼠,第0、5天免疫组肿瘤复发数少于第5,8天免疫组(1/10,4/10 vs 8/10,7/10).结论 PK-HPV-ad5疫苗能诱导小鼠产生特异性的免疫应答,对抗肿瘤复发有治疗潜力.
목적 평개HPV16 E6E7적복제결함형중조5형선병독(PK-HPV-ad5)치료성역묘대실험소서면역응답화항종류적생물학효응.방법 사용기인중조기술구건PK-HPV-ad5역묘,병통과소서면역시험,검측소서총항체화특이성IFNγ,동시장조모소서분성역묘조화대조조,분별대기진행억류시험、TC-1종류세포도전시험화종류절제후방복발시험.결과 HPV16 E6E7유도적총항체제12천적수평상대교고(1:400～1:600);3비차역묘특이성IFNγ재제14천여대조조비교분별승고8.6、5.9화8.9배,차이유통계학의의(t=15.721、6.967화14.342,P균＜0.01).억류시험표명역묘제량위107IU/지시소서종류생장솔위0,여대조조비교차이유통계학의의(학절개솔법,P＜0.01),3비차역묘험증유효제량위107IU/지시종류억제솔가체80％(8/10)이상.TC-1종류세포도전시험결과현시:소서선접충역묘능인기특이성적면역응답,병능보호90％(9/10)적소서면수TC-1종류세포적공격;종류절제후방지복발시험제시재주사상동제량역묘시,대104개/지화105개/지종류세포조모소서,제0、5천면역조종류복발수소우제5,8천면역조(1/10,4/10 vs 8/10,7/10).결론 PK-HPV-ad5역묘능유도소서산생특이성적면역응답,대항종류복발유치료잠력.
Objective To evaluate the immune responses and anti-tumor effects of replication-deficient recombinant adenovirus-5 vector vaccine of human papillomavirus type 16 E6E7 as a theraputic vaccine (PK-HPV-ad5) in mouse models.Methods PK-HPV-ad5 vaccine was constructed by gene recombination technique.HPV16E6E7 total antibody and specific IFNγ of the vaccine were detected by mouse immune experiment.The model mice were divided into vaccine group and control group,and were used for anti-tumor test,TC-1 tumor cell challenge test and evaluation of tumor excision combined with vaccine to prevent tumor recurrence.Results HPV16 E6E7 total antibody increased to a comparatively high level (1:400-1:600) on d12 after vaccination.The specific IFNγin 3 batches of vaccine(20121201,20121202 and 20121203) were 8.6,5.9 and 8.9 times of those in control group on d14 after vaccination,respectively,with significant differences(t=15.721,6.967 and 14.342,P all＜0.01).The anti-tumor test showed that there was no tumor grew in vaccine group at the dosage of 107 IU per mouse,with a significant difference to control group(definite probability methods,P＜0.01).The anti-tumor test of 3 batches vaccines conformed that the tumor suppression rate was over 80％(8/10) at dosage of 107 IU per mouse.TC-1 tumor cell challenge test showed that the mice had immune response after vaccination,and 90％ (9/10)mice escaped TC-1 cell attack.Tumor recurrence test suggested that with the same vaccination,the number of recurrent tumor lesions in model mice of 104 tumor cells per mouse inoculated on d0 and d5 were fewer than that in model mice of 105 tumor cells per mouse on d5 and d8 (1/10,4/10 vs 8/10,7/10).Conclusions PK-HPV-ad5 vaccine can elicit mice producing specific immune response.It has potential effect against tumor recurrence.