Moore Richard A, Santos Elmer B, Nicholls Philip K, White Kate L, Anderson Davina M, Lloyd Andrew, Topley Peter, Romanos Michael, Thomsen Lindy, Parmar Vanita, Walcott Sarah, Gough Gerald W, Stanley Margaret A
Department of Pathology, University of Cambridge, Tennis Court Road, Cambridge CB2 1QP, United Kingdom.
Virology. 2002 Dec 20;304(2):451-9. doi: 10.1006/viro.2002.1726.
DNA plasmids encoding the open reading frames of canine oral papillomavirus (COPV) nonstructural early genes E1, E2, or E7 protein were delivered into both oral mucosal and cutaneous epithelial sites in beagle dogs using particle-mediated immunotherapeutic delivery (PMID) technology. Control dogs were vaccinated with plasmid encoding either hepatitis B virus surface antigen (HBVs) or COPV L1. Using a prophylactic immunisation protocol, a priming dose of plasmid DNA was followed by a booster dose 6 weeks later. Four weeks after boost, all dogs were challenged with infectious COPV particles. Following viral challenge, as shown previously (M. A. Stanley et al., 2001, Vaccine 19, 2783-2792), mucosal papillomas developed in the negative-control HBVs vaccinated dogs, but all animals in the COPV L1 group were fully protected from disease development. In the early gene-vaccinated groups five of six in the E1-vaccinated dogs, two of six in E2-vaccinated dogs, and three of six in the E7-vaccinated beagles developed oral papillomas. Compared to the HBVs negative-control group the oral papillomas that did develop in the early-gene vaccinated beagles were significantly smaller, shorter in duration, and fewer in number. Taken together the disease burden was markedly reduced and this was statistically significant. In a second experiment one group of animals was vaccinated with plasmid encoding the wild-type COPV E1 gene, and a separate group was vaccinated with plasmid encoding a synthetic codon-optimised COPV E1 gene sequence. None of the codon-optimised E1-vaccinated animals developed papillomas at any challenge site. However, all animals vaccinated with wild-type E1 had papillomas. These data suggest that immunisation by PMID with papillomavirus early genes can significantly impact upon subsequent disease development and that full protection can be achieved using improved vectors encoding codon-optimised gene sequences perhaps emphasizing the importance of antigen load in the generation of protective responses to papillomavirus proteins.
使用粒子介导免疫治疗递送（PMID）技术，将编码犬口腔乳头瘤病毒（COPV）非结构早期基因E1、E2或E7蛋白开放阅读框的DNA质粒递送至比格犬的口腔黏膜和皮肤上皮部位。对照犬接种编码乙型肝炎病毒表面抗原（HBVs）或COPV L1的质粒。采用预防性免疫方案，先给予一剂质粒DNA作为初始剂量，6周后再给予加强剂量。加强免疫4周后，所有犬均用感染性COPV颗粒进行攻击。如先前所示（M. A. Stanley等人，2001年，《疫苗》19卷，2783 - 2792页），接种阴性对照HBVs的犬出现黏膜乳头瘤，但COPV L1组的所有动物均完全免受疾病发展影响。在早期基因疫苗接种组中，接种E1的犬6只中有5只、接种E2的犬6只中有2只、接种E7的比格犬6只中有3只出现口腔乳头瘤。与HBVs阴性对照组相比，早期基因疫苗接种的比格犬中出现的口腔乳头瘤明显更小、持续时间更短且数量更少。总体而言，疾病负担显著减轻，且具有统计学意义。在第二项实验中，一组动物接种编码野生型COPV E1基因的质粒，另一组接种编码合成密码子优化的COPV E1基因序列的质粒。接种密码子优化E1的动物在任何攻击部位均未出现乳头瘤。然而，所有接种野生型E1的动物均出现乳头瘤。这些数据表明，通过PMID用乳头瘤病毒早期基因进行免疫可显著影响后续疾病发展，并且使用编码密码子优化基因序列的改良载体可实现完全保护，这可能强调了抗原负载在产生针对乳头瘤病毒蛋白的保护性反应中的重要性。
