含密码子优化型HPV16 L1基因重组腺病毒的构建及其免疫效果的研究

目的 构建含密码子优化型HPV16L1基因的重组腺病毒,对其经不同接种途径所诱导的系统性及黏膜免疫效果进行研究.方法 使用Admax系统包装重组腺病毒,纯化的腺病毒以不同方式免疫C57BL/6小鼠,间接ELISA及体外中和实验检测免疫小鼠血清及阴道分泌物中的特异性抗体.结果 重组腺病毒滴鼻接种可同时诱导特异性的系统性及黏膜免疫反应,重组腺病毒肌注免疫仅能诱导系统性免疫反应,而阴道黏膜接种不能有效诱导系统性及黏膜免疫反应.结论 成功构建了含密码子优化型HPV 16 L1基因的重组腺病毒,重组腺病毒肌注可诱导高滴度的血清中和抗体,滴鼻接种可同时诱导特异性的系统性及黏膜免疫反应.

Construction and immunological evaluation of recombinant adenovirus containing codon-modified HPV 16 L1 gene

OBJECTIVE: To construct recombinant adenovirus containing codon-modified HPV16L1 gene, and evaluate systemic and mucosal immunological responses induced after immunization with the recombinant virus. METHODS: The recombinant adenovirus rAd-mod.HPV16L1 was constructed by Admax kit. The C57 BL/6 mice were immunized by purified rAd-mod.HPV16L1 through different inoculation routes. The immunological effect was evaluated by testing the specific neutralizing antibodies in sera and vaginal secretions of immunized mice through indirect ELISA and neutralization assay based HPV pseudovirus. RESULTS: The result showed that intramuscular immunization could induce good systemic immunity, but the mucosal immunity was too weak, and immunization via intranasal route could induce satisfactory immunity both in sera and vaginal secretions, while intravaginal immunization failed to induce any specific immunological responses either in sera or vaginal secretions. CONCLUSION: The recombinant adenovirus containing codon- modified HPV16L1 gene was successfully constructed. Immunization through intranasal route could induce satisfactory immunity both in sera and vaginal secretions, while intramuscular immunization could only induce high titer of neutralizing antibodies in sera.