Human Papillomavirus type 16 (HPV16) has been associated with a subset of head and neck cancers. Two HPV encoded oncogenic proteins, E6 and E7, are important for the malignant progression of HPV-associated cancers. A spontaneous HPV16 E6/E7-expressing oral tumor model in human HLA-A2 (AAD) transgenic mice will be important for the development of therapeutic HPV vaccines for the control of HPV-associated head and neck cancers. In the current studies, we characterized the HLA-A2 restricted HPV16 E7-specific CD8 + T cell mediated immune responses in the HLA-A2 (AAD) transgenic mice using a therapeutic naked DNA vaccine encoding calreticulin (CRT) linked to a mutated E7(N53S). We also employed oncogenic DNA plasmids that encoded HPV16E6/E7/Luc, NRasG12V, and sleeping beauty transposase for the transfection into the submucosal of oral cavity of the transgenic mice with electroporation to create a spontaneous oral tumor. Furthermore, we characterized the therapeutic antitumor effects of CRT/E7(N53S) DNA vaccine using the spontaneous HPV16 E6/E7-expressing oral tumor model in HLA-A2 (AAD) transgenic mice. We found that CRT/E7(N53S) DNA vaccine primarily generated human HPV16 E7 peptide (aa11-20) specific CD8 + T cells, as compared to the wild-type CRT/E7 vaccine, which primarily generated murine H-2Db restricted E7 peptide (aa49-57) specific CD8 + T cell responses. We also observed transfection of the oncogenic DNA plasmids with electroporation generated spontaneous oral tumor in all of the injected mice. Additionally, treatment with CRT/E7(N53S) DNA vaccine intramuscularly followed by electroporation resulted in significant antitumor effects against the spontaneous HPV16 E6/E7-expressing oral tumors in HLA-A2 (AAD) transgenic mice. Taken together, the data indicated that the combination of HPV16 E6/E7-expressing DNA, NRasG12V DNA and DNA encoding sleeping beauty transposase is able to generate spontaneous oral tumor in HLA-A2 (AAD) transgenic mice, which can be successfully controlled by treatment with CRT/E7(N53S) DNA vaccine. The translational potential of our studies are discussed.

中文翻译：

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用治疗性 HPV DNA 疫苗控制 HLA-A2 (AAD) 转基因小鼠中自发的 HPV16 E6/E7 表达口腔癌

人乳头瘤病毒 16 型 (HPV16) 与一部分头颈癌有关。两种 HPV 编码的致癌蛋白 E6 和 E7 对 HPV 相关癌症的恶性进展很重要。在人 HLA-A2 (AAD) 转基因小鼠中自发表达 HPV16 E6/E7 的口腔肿瘤模型对于开发用于控制 HPV 相关头颈癌的治疗性 HPV 疫苗具有重要意义。在目前的研究中，我们使用编码钙网蛋白 (CRT) 的治疗性裸 DNA 疫苗与突变的 E7 ( N53S)。我们还使用了编码 HPV16E6/E7/Luc、NRasG12V、睡美人转座酶用于转染转基因小鼠口腔黏膜下层，通过电穿孔产生自发性口腔肿瘤。此外，我们在 HLA-A2 (AAD) 转基因小鼠中使用自发表达 HPV16 E6/E7 的口腔肿瘤模型表征了 CRT/E7(N53S) DNA 疫苗的治疗性抗肿瘤作用。我们发现，与主要产生鼠类 H-2Db 限制性 E7 肽的野生型 CRT/E7 疫苗相比，CRT/E7(N53S) DNA 疫苗主要产生人 HPV16 E7 肽 (aa11-20) 特异性 CD8 + T 细胞(aa49-57) 特异性 CD8 + T 细胞反应。我们还观察到用电穿孔转染致癌 DNA 质粒会在所有注射小鼠中产生自发性口腔肿瘤。此外，在 HLA-A2 (AAD) 转基因小鼠中，用 CRT/E7(N53S) DNA 疫苗肌内注射和电穿孔对自发表达 HPV16 E6/E7 的口腔肿瘤产生显着的抗肿瘤作用。综上所述，数据表明，表达 HPV16 E6/E7 的 DNA、NRasG12V DNA 和编码睡美人转座酶的 DNA 的组合能够在 HLA-A2 (AAD) 转基因小鼠中产生自发性口腔肿瘤，并且可以通过治疗成功控制用 CRT/E7(N53S) DNA 疫苗。讨论了我们研究的转化潜力。NRasG12V DNA 和编码睡美人转座酶的 DNA 能够在 HLA-A2 (AAD) 转基因小鼠中产生自发性口腔肿瘤，可以通过 CRT/E7(N53S) DNA 疫苗治疗成功控制。讨论了我们研究的转化潜力。NRasG12V DNA 和编码睡美人转座酶的 DNA 能够在 HLA-A2 (AAD) 转基因小鼠中产生自发性口腔肿瘤，可以通过 CRT/E7(N53S) DNA 疫苗治疗成功控制。讨论了我们研究的转化潜力。