含HPV16L1基因重组腺病毒和1型重组AAV载体联合免疫效果的研究

Immune response induced by recombinant adenovirus combined with recombinant adeno-associated virus type 1 containing HPV16 L1 gene

目的对表达HPV16L1抗原的重组腺病毒及1型重组AAV载体联合免疫效果进行研究。方法分别构建含密码子优化型HPV16L1基因重组腺病毒rAd-mod．HPV16L1及1型重组AAV载体rAAV1-mod．HPV 16L1，将纯化的重组AAV病毒载体以肌注及滴鼻途径单独及联合免疫C57BL／6小鼠，使用体外中和实验检测各组小鼠血清中特异性中和抗体。结果rAAV1-mod．HPV16L1单独及与rAd-mod．HPV16L1联合肌注可诱导高滴度的血清中和抗体，在初免后第16周抗体滴度显著高于其他免疫组，联合肌注组诱导的抗体滴度高于单独肌注组；重组病毒联合滴鼻虽能产生一定的免疫加强作用，但抗体滴度仍显著低于rAAV1-mod．HPV16L1单独及联合肌注组。结论型重组AAV载体联合重组腺病毒以初免-加强模式肌注町诱导更高滴度的血清中和抗体。

Objective To evaluate the immune potency of recombinant adenovirus combined with rAAV1 vector expressing HPV16L1 protein in mice. Methods The rAdV and rAAV1 vector containing codon-modified HPV16L1 gene was constructed using Admax and AAVmax packaging system respectively. C57 BL/6 mice were immunized with purified rAdV and rAAV1 vector through intramuscular and intranasal inoculation routes, and the titer of neutralizing antibody was determined by neutralization assay based HPV16 pseudovirus. Results Intramuscular immunization by rAAVl-mod. HPV16L1 or combined with rAd-mod. HPV16L1 can induce higher titer of neutralizing antibody in serum than that of other groups. The titer of neutralizing antibody of intranasal groups is significantly lower than that of intramuscular group, although the prime-boost strategy, using in intranasal group was effective to enhance the specific humoral immunity. Conclusion The rAAVl-mod. HPV16L1 combined with tAd-rood. HPV16L1 can induce higher titer of neutralizing antibody in serum through intramuscular route than that of other groups at the 16th week after the first immunization.