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Characterization of HPV18 E6-specific T cell responses and establishment of HPV18 E6-expressing tumor model

机译：HPV18 E6特异性T细胞应答的表征及HPV18 E6表达肿瘤模型的建立

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Human papillomavirus (HPV) has been identified as the primary etiologic factor of cervical cancer, and subsets of anogenital and oropharyngeal cancers. HPV18 is the second most prevalent high-risk HPV type after HPV16. Furthermore, HPV18 is responsible for approximately 12% of cervical squamous cell carcinoma and 37% of cervical adenocarcinoma cases worldwide. In this study, we aimed to characterize the HPV18-E6-specific epitope and establish an HPV18 animal tumor model to evaluate the E6-specific immune response induced by our DNA vaccine. We vaccinated naive C57BL/6 mice with a prototype DNA vaccine, pcDNA3-HPV18-E6, via intramuscular injection followed by electroporation, and analyzed the E6-specific CD8+ T cell responses by flow cytometry using a reported T cell epitope. We then characterized the MHC restriction element for the characterized HPV18-E6 epitope. Additionally, we generated an HPV18-E6-expressing tumor cell line to study the antitumor effect mediated by E6-specific immunity. We observed a robust HPV18-E6aa67-75 peptide-specific CD8+ T cell response after vaccination with pcDNA3-HPV18-E6. Further characterization demonstrated that this epitope was mainly restricted by H-2K(b), but was also weakly presented by HLA-A*0201, as previously reported. We observed that vaccination with pcDNA3-HPV18-E6 significantly inhibited the growth of HPV18-E6-expressing tumor cells, TC-1/HPV18-E6, in mice. An antibody depletion study demonstrated that both CD4+ and CD8+ T cells are necessary for the observed antitumor immunity. The characterization of HPV18-E6-specific T cell responses and the establishment of HPV18-E6-expressing tumor cell line provide infrastructures for further development of HPV18-E6 targeted immunotherapy. (C) 2017 Elsevier Ltd. All rights reserved.
机译：人乳头瘤病毒（HPV）已被鉴定为宫颈癌的主要病因因素，以及吞咽和口咽癌症的亚群。 HPV18是HPV16后的第二个最普遍的高风险HPV类型。此外，HPV18负责大约12％的宫颈鳞状细胞癌和全球颈椎腺癌病例的37％。在这项研究中，我们旨在表征HPV18-E6特异性表位并建立HPV18动物肿瘤模型，以评估我们的DNA疫苗诱导的E6特异性免疫应答。我们通过肌内注射用原型DNA疫苗接种幼稚的C57BL / 6小鼠，然后通过肌内喷射，然后通过报告的T细胞表位通过流式细胞仪分析E6特异性CD8 + T细胞应答。然后，我们表征了表征HPV18-E6表位的MHC限制元素。另外，我们生成了表达HPV18-E6表达肿瘤细胞系，以研究由E6特异性免疫介导的抗肿瘤效应。在用PCDNA3-HPV18-E6接种后，观察到稳健的HPV18-E6AA67-75肽特异性CD8 + T细胞响应。进一步表征证明该表位主要受H-2K（B）限制，但是如前所述，HLA-A * 0201也弱呈效率。我们观察到使用PCDNA3-HPV18-E6的疫苗接种显着抑制了小鼠中HPV18-E6表达肿瘤细胞TC-1 / HPV18-E6的生长。抗体耗尽研究表明，CD4 +和CD8 + T细胞是观察到的抗肿瘤免疫所必需的。 HPV18-E6特异性T细胞应答的表征和HPV18-E6表达肿瘤细胞系的建立提供了用于进一步发展HPV18-E6靶向免疫疗法的基础设施。 （c）2017 Elsevier Ltd.保留所有权利。

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