Nieto Karen, Kern Andrea, Leuchs Barbara, Gissmann Lutz, Müller Martin, Kleinschmidt Jürgen A
Infection and Cancer Research Program, German Cancer Research Center, Heidelberg, Germany.
Antivir Ther. 2009;14(8):1125-37. doi: 10.3851/IMP1469.
Cervical cancer is the second most frequent cancer among woman worldwide and is considered to be caused by infection with high-risk papilloma viruses. Genetic immunization using recombinant adeno-associated virus (rAAV) vectors has shown great promise for vaccination against human papillomavirus (HPV) infections.
rAAV5, -8 and -9 vectors expressing an HPV16 L1/E7 fusion gene were generated and applied intranasally for combined prophylactic and therapeutic vaccination of mice.
The rAAV5 and the rAAV9 vectors showed efficient induction of both humoral and cellular immune responses, whereas rAAV8 failed to immunize mice by the intranasal route. The L1-specific immune response evoked by expression of the L1/E7 fusion gene, however, was lower than that evoked by expression of the L1 antigen alone. This deficiency could be compensated by application of Escherichia coli heat-labile enterotoxin or monophsphoryl lipid as adjuvant upon vaccination with rAAV5-L1/E7. Coimmunization of rAAV9-L1/E7 with rAAV5-L1 or boosting of rAAV9-L1/E7 with rAAV5-L1 strongly increased L1-specific neutralizing antibody titres to levels above those achieved by vaccination with vectors expressing L1 alone. Both vectors elicited a vibrant cytotoxic T-lymphocyte response against L1 or E7. Nasal immunization with rAAV5 or rAAV9 was superior to vaccination with HPV16-L1 virus-like particles (VLPs) or HPV16-L1/E7 CVLPs with respect to humoral and cellular immune responses. Vaccination with the rAAV vectors led to a significant protection of animals against a challenge with different HPV tumour cell lines.
Our results show that rAAV5 and rAAV9 vectors are promising candidates for a non-invasive nasal vaccination strategy.
宫颈癌是全球女性中第二常见的癌症，被认为是由高危乳头瘤病毒感染引起的。使用重组腺相关病毒（rAAV）载体进行基因免疫在预防人乳头瘤病毒（HPV）感染的疫苗接种方面显示出巨大潜力。
构建表达HPV16 L1/E7融合基因的rAAV5、-8和-9载体，并经鼻应用于小鼠的联合预防性和治疗性疫苗接种。
rAAV5和rAAV9载体均能有效诱导体液免疫和细胞免疫反应，而rAAV8经鼻途径未能使小鼠产生免疫。然而，L1/E7融合基因表达所引发的L1特异性免疫反应低于单独表达L1抗原所引发的反应。在用rAAV5-L1/E7接种疫苗时，应用大肠杆菌不耐热肠毒素或单磷酰脂质作为佐剂可弥补这一缺陷。rAAV9-L1/E7与rAAV5-L1共同免疫或用rAAV5-L1加强免疫rAAV9-L1/E7，可使L1特异性中和抗体滴度大幅提高，超过单独使用表达L1的载体接种所达到的水平。两种载体均引发了针对L1或E7的强烈细胞毒性T淋巴细胞反应。在体液免疫和细胞免疫反应方面，用rAAV5或rAAV9进行鼻内免疫优于用HPV16-L1病毒样颗粒（VLP）或HPV16-L1/E7嵌合病毒样颗粒（CVLP）进行接种。用rAAV载体接种疫苗可显著保护动物免受不同HPV肿瘤细胞系的攻击。
我们的结果表明，rAAV5和rAAV9载体是无创鼻内疫苗接种策略的有前景的候选者。
