Study of a DNA vaccine of the papillomavirus

在之前的研究中，我们建立了一个高重复性的致癌模型，将DMBA应用与仓鼠舌粘膜的物理损伤相结合。组织学上，这些病变表现出乳头瘤病毒感染的病理改变，如发育不良细胞的嗜空细胞增多，一些发育不良细胞或癌细胞的细胞核中乳头瘤病毒属特异性抗原（GSA）阳性。此外，在这些细胞的细胞核中检测到类似乳头瘤病毒的病毒颗粒，其直径为50-55 nm。利用这种动物模型，我们通过对这种病毒基因组的分子克隆和测序证明了一种新型仓鼠乳头瘤病毒的存在。全序列分析证实该病毒为一种新型乳头瘤病毒，命名为仓鼠口腔乳头瘤病毒（HOPV）。此外，我们用原位杂交技术在仓鼠舌上皮发育不良和恶性病变的细胞核中检测到HOPV。更多的计算机辅助系统发育分析显示HOPV在基因组的不同区域与其他乳头瘤病毒具有不同的同源性。该病毒基因组全核苷酸序列为7647bp， G+C含量为46.6%。这个大小与MnPV相似（7687bp）。该基因组在一条链上有开放阅读框（orf）， orf的顺序为：E6、E7、E1、E2、L2和L1。ORF E2包含较小的ORF E4。ORF H5缺失。这些克隆基因组不同于其他啮齿动物乳头瘤病毒，如MmPV和MnPV，也不同于先前测序的动物和人类乳头瘤病毒基因组。我们评估了DNA疫苗的预防作用。每隔3周肌肉注射2次HOPV E6/E7/L1质粒DNA。用这些裸pdna观察到保护作用。因此，HOPV感染似乎是研究乳头瘤病毒相关粘膜癌变、疫苗预防和恶性转化预防或治疗研究的良好模型。少

In a previous investigation, we developed a highly reproducible carcinogenesis model by combining DMBA application with physical wounding of the hamster lingual mucosa. Histologically, these lesions showed pathognomonic changes indicative of papillomavirus infection, such as koilocytosis of dysplastic cells, and the nuclei of some dysplastic or cancerous cells were positive for papillomavirus genus-specific antigen (GSA). Furthermore, viral particles resembling papillomavirus, whose size was 50-55 nm in diameter, have been detected in the nuclei of these cells. Using this animal model, we demonstrated the presence of a novel hamster papillomavirus by the molecular cloning and sequencing of this viral genome. The genome was confirmed by complete sequence analysis to be a novel papillomavirus which we designated the hamster oral papillomavirus (HOPV). Moreover, we detected HOPV in the nuclei of dysplastic and malignant lesions in hamster lingual epithelium using in situ hybridization. Co … More mputer-assisted phylogenetic analysis of HOPV showed a differing homology with other papillomaviruses in different regions of the genomes. The complete nucleotide sequence of this viral genome consists of 7647bp with a G+C content of 46.6%. This size is similar to that of MnPV(7687bp). This genome has open reading frames (ORFs) on one strand showing the usual order : ORFs E6, E7, E1, E2, L2 and L1. ORF E2 contains the small ORF E4. ORF H5 is missing. These cloned genomes are different from those of other rodent papillomaviruses, such as MmPV and MnPV, and also from previously sequenced animal and human papillomavirus genomes.We evaluated a prophylactic effect of DNA vaccination. Hamsters were injected intramusccularly with HOPV E6/E7/L1 plasmid DNA two times at 3-week intervals. The protective effect was observed with those naked pDNAs. Therefore, HOPV infection seems to be a good model for the study on mucosal carcinogenesis associated with papillomavirus and for studies on vaccine prophylaxis and the prevention or treatment of malignant conversion. Less