重组腺病毒及其基因组裸DNA免疫效果差异的初步分析  

Immunization with recombinant adenoviral genome and recombinant adenovirus induces effective immune reaction by direct muscle inoculation

作　　者：卞继峰[1] 于修平[1] 穆玉兰[2] 胡海燕[1] 耿昭[1] 卢翌[1] 王伟[1] 

机构地区：[1]山东大学医学院分子生物学实验中心,山东济南250012 [2]山东省立医院妇产科,山东济南250021

出　　处：《山东大学学报（医学版）》2005年第6期461-463,499,共4页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金资助课题(30170045;30200246);霍英东青年教师基金资助课题(91044)

摘　　要：目的:研究重组腺病毒及其基因组裸DNA免疫效果的差异,探讨重组腺病毒疫苗免疫接种的新途径。方法:构建人乳头瘤病毒重组腺病毒rAd5-L1E7,用TCID50微量滴定法测定病毒滴度。将0.1ml滴度为10-4重组腺病毒和10μg重组腺病毒基因组DNA分别肌肉注射免疫BALB/c小鼠,在接种后第1～5周分离小鼠血清。以HPV16L1E7重组蛋白为抗原,用ELISA方法检测血清HPV16特异性抗体水平。通过T细胞增殖试验和检测脾细胞IFN-γ水平确定疫苗诱导的细胞免疫,并分析二者之间的差异。结果:重组腺病毒及其基因组裸DNA免疫接种均能有效地诱导体液和细胞免疫,抗体达到高峰的时间不同,二者在第3和第4周血清抗体IgG水平差异有统计学意义(P<0.05)。T淋巴细胞增殖试验结果和脾细胞分泌IFN-γ水平差异无统计学意义。结论:重组腺病毒基因组裸DNA接种能够有效地诱导细胞和体液免疫应答,是腺病毒疫苗接种的一种新途径。Objective: To study the possibility of a new inoculation pathway of recombinant adenovirus vaccine application. Methods: Recombinant adenovirus of human papillomavirus type 16 and its genomic DNA were used as material. rAd5-L1E7 was inoculated through oral and nasal. The rAd5-L1E7 genome DNA was extracted and purified, then BALB/c mice were immunized intramuscularly with rAd5-L1E7 and its genomic DNA separately. The serum was collected from week 1 to week 5 postinoculation, HPV16 specific IgG was measured with HPV16 L1E7-based ELISA. The CMI effects were measured using T lymphocyte proliferation test and IFN-γ level secreted by HPV16 L1E7 protein-pulsed spleen cells. Results: Recombinant adenovirus and naked rAd5-l1E7 genomic DNA was able to induce effective humoral and cellular immune reaction. No significant difference was found about serum IgG level and data of T lymphocyte proliferation test and IFN-γ induced by virus and naked genomic DNA immunization. Conclusion: Direct inoculation of naked recombinant adenovirus genome DNA is an alternative and effective pathway.

关 键 词：重组腺病毒 DNA重组 乳头瘤病毒 病毒疫苗 小鼠 近交BALB C 

分 类 号：R373.9[医药卫生—病原生物学] Q782[医药卫生—基础医学]