HPV58型复合DNA疫苗抗肿瘤免疫效果的初步观察  

Preliminary observation on antitumor effect of HPV58 composite DNA vaccine

作　　者：王鹤[1] 于继云[2] 李力[1] 

机构地区：[1]广西医科大学附属肿瘤医院妇瘤科,南宁530021 [2]军事医学科学院基础医学研究所转化医学研究室

出　　处：《中华妇产科杂志》2013年第7期523-527,共5页Chinese Journal of Obstetrics and Gynecology

基　　金：广西壮族自治区卫生厅重点科研课题（重200970）

摘　　要：目的 初步观察PVAX1-HPV58mE6E7FcGB复合DNA疫苗的抗肿瘤免疫效果.方法 在检测该复合疫苗的免疫效果前,自行构建了稳定表达HPV58E6E7融合基因的B16-HPV58E6E7成瘤细胞系,表达并纯化了HPV58E6E7-GST融合蛋白作为抗原.将该疫苗免疫C57BL/6小鼠（免疫组）后,通过抗肿瘤移植保护实验和肿瘤生长抑制实验观察该疫苗对小鼠负荷的B16-HPV58E6E7移植瘤的防治效果；通过特异性抗体检测、酶联免疫斑点检测、特异性杀伤实验观察该疫苗在被免疫小鼠体内诱发的体液和细胞免疫反应.以单纯抗原PVAX1-HPV58mE6E7 Fc免疫小鼠作为单纯抗原组.结果 抗肿瘤移植保护实验显示,经PVAX1-HPV58mE6E7FcGB疫苗免疫的小鼠成瘤率仅为9/15,但成瘤时间最长[为（13.6±1.7）d],且肿瘤生长最慢.在肿瘤生长抑制实验中,疫苗组小鼠的肿瘤生长抑制率达81.4％.体液免疫检测显示,该疫苗和单纯抗原均能在小鼠体内诱发特异性抗体,最高滴度分别为1∶25 600和1∶12 800,两组比较,差异无统计学意义（P＞0.05）；但在细胞免疫检测中,疫苗组激活的特异性T淋巴细胞数[（219±34）个/4×105个脾淋巴细胞]约为单纯抗原组[（55±25）个/4×105个脾淋巴细胞]的4倍,两组比较,差异有统计学意义（P＜0.05）；且该疫苗诱发的特异性针对B 16-HPV58 E6 E7细胞的杀伤率（最高为43.3％）也明显高于单纯抗原组（杀伤率最高为31.3％）,两组比较,差异有统计学意义（P＜0.05）.结论 PVAX1-HPV58mE6E7FcGB复合DNA疫苗能有效激发特异性的体液和细胞免疫反应,显著抑制B16-HPV58E6E7肿瘤细胞的生长,在细胞免疫上优于单纯抗原,可作为HPV58阳性相关肿瘤及其癌前病变免疫治疗的候选疫苗.Objective To initially observe the antitumor immune of PVAX1-HPV58mE6E7FcGB composite DNA vaccine.Methods Before detecting immune effect of the vaccine,the B16-HPV58E6E7 tumor cell line was built which could steadily express HPV58E6E7 fusion gene.Then,HPV58E6E7-GST fusion protein as an antigen was expressed and purified.Before or after immunized with the vaccine,the C57BL/6 mice were challenged by B16-HPV58E6E7 cells.Anti-tumor transplantation and tumor growth inhibition experiment were performed to observe prevention and treatment effects on the vaccine.Specific humoral and cellular immune responses in the immunized mice were detected by ELISA,enzyme linked immunospot assay （ELISPOT） and cytotoxic T lymphocyte （CTL） method.Results In the anti-tumor transplantation experiment,tumor formation rate was only 9/15 in the mice which were immunized by PVAX1-HPV58mE6E7FcGB vaccine,time before tumor formation was the longest [（13.6 ± 1.7） days] and tumor growth was the slowest in the vaccine group.In tumor growth inhibition experiment,inhibition rate reached 81.4% in the vaccine group.Except tumor formation rate,all data in the vaccine group was superior to the pure antigen PVAX1-HPV58mE6E7Fc group （P 〈 0.05）.Humoral immune effect showed that both the vaccine and the pure antigen could induce specific antibody in the immunized mice,and the highest titer were 1 ∶ 25600 and 1 ∶ 12800,respectively.Although there was not significant difference of antibody titer between the vaccine and the pure antigen group （P 〉 0.05）,the number of activated T cells in the vaccine group was almost four times as that in the pure antigen group [（219 ±34）/4 × 105 spleen lymphocytes versus （55 ±25）/4 × 105 spleen lymphocytes,P 〈 0.05],and the highest specific CTL that vaccine induced was significantly higher than that of pure antigen （43.3% versus 31.3%,P 〈 0.05）.Conclusions Humoral and cellular immune response could be effectively stimulated by PVAX1-HPV58mE6E7FcGB composite DNA vaccine.Growth of

关 键 词：乳头状瘤病毒科 癌基因蛋白质类 病毒性 衣壳蛋白质类 DNA疫苗 免疫活性 小鼠 近交系 

分 类 号：R392[医药卫生—免疫学]