表达人乳头瘤病毒16型E6E7重组腺病毒疫苗对小鼠的免疫和抗肿瘤效应  

Immune responses of recombinant adenovirns-5 vector vaccine of human papillomavirus type 16 E6E7 and its anti-tumor effects in mice

作　　者：吴洁[1] 陈刚[1] 金素凤 高孟[1] 庄昉成[1] 李剑波[1] 姜云水[1] 毛子安 

机构地区：[1]浙江省医学科学院病毒病研究所,杭州310013 [2]浙江普康生物技术股份有限公司,杭州310053

出　　处：《国际流行病学传染病学杂志》2014年第6期368-373,共6页International Journal of Epidemiology and Infectious Disease

基　　金：浙江省自然科学基金（LY14C080001）;浙江省医学生物工程疫苗研发重点实验室（2008F3022）;杭州市科技局重大科技创新项目（20112313A37）

摘　　要：目的评价HPVl6E6E7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—ad，疫苗，并通过小鼠免疫试验，检测小鼠总抗体和特异性IFNl，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HPV16E6E7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IFNl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10^7IU／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10^7IU／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10^4个，只和10^5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（1／10，4／10VS8／10，7／10）。结论PK—HPV—ads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。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-ads） 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 IFN/in 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 10^7 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 10^7 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 10^4 tumor cells per mouse inoculated on dO 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.

关 键 词：乳头状瘤病毒疫苗 腺病毒疫苗 免疫应答 抗肿瘤作用 

分 类 号：R73-36[医药卫生—肿瘤]