Objectives

Human papillomavirus infection (HPV) is the most common viral infection which is causes of cervical, penal, vulvar, anal and, oropharyngeal cancer. E7 protein of HPV is a suitable target for induction of T cell responses and controlling HPV-related cancer. The aim of the current study was to designed and evaluated a novel fusion protein containing the different E7 proteins of the HPV 16, 18, 6 and 11, linked to the cell-penetrating peptide HIV-1 Tat 49–57, in order to improve cytotoxic immune responses in in-vitro and in-vivo.

Results

In this study whole sequence of HPV16,18,6,11 E7-Tat (47–57) and HPV16,18,6,11 E7 cloned into the vector and expressed in E. coli (BL21). The purified protein was confirmed by SDS page and western blotting and then injected into the C57BL/6 mice. The efficiency of the fusion protein vaccine was assessed by antibody response assay, cytokine assay (IL-4 and IFN-γ), CD + 8 cytotoxicity assay and tumor challenge experiment. Result showed that fusion proteins containing Adjuvant (IFA,CFA) could express higher titer of antibody. Also, we showed that vaccination with E7-Tat and, E7-Tat-ADJ induced high frequencies of E7-specific CD8 + T cells and CD107a expression as well as IFN-γ level and enhanced long-term survival in the therapeutic animal models.

Conclusion

Our finding suggested that this novel fusion protein vaccine was able to induce therapeutic efficacy and immunogenicity by improving CD8 + T cell in TC-1 tumor bearing mice; so this vaccine may be appreciated for research against HPV and tumor immunotherapies.

中文翻译：

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一种新型重组蛋白疫苗，含有与 HIV-1 Tat (47-57) 相关的 HPV16、18、6、11 E7 的不同 E7 蛋白，可改善细胞毒性免疫反应

目标

人乳头瘤病毒感染 (HPV) 是最常见的病毒感染，可导致宫颈癌、阴茎癌、外阴癌、肛门癌和口咽癌。HPV的E7蛋白是诱导T细胞反应和控制HPV相关癌症的合适靶标。本研究的目的是设计和评估一种新型融合蛋白，该融合蛋白含有 HPV 16、18、6 和 11 的不同 E7 蛋白，与细胞穿透肽 HIV-1 Tat 49-57 相连，以改善体外和体内的细胞毒性免疫反应。

结果

在这项研究中，将 HPV16、18、6、11 E7-Tat (47-57) 和 HPV16、18、6、11 E7 的整个序列克隆到载体中并在大肠杆菌 (BL21) 中表达。纯化的蛋白质通过 SDS 页和蛋白质印迹确认，然后注射到 C57BL/6 小鼠中。通过抗体反应测定、细胞因子测定（IL-4和IFN-γ）、CD+8细胞毒性测定和肿瘤攻击实验评估融合蛋白疫苗的效率。结果表明，含有佐剂（IFA，CFA）的融合蛋白可以表达更高滴度的抗体。此外，我们发现用 E7-Tat 和 E7-Tat-ADJ 接种疫苗可诱导高频率的 E7 特异性 CD8 + T 细胞和 CD107a 表达以及 IFN-γ 水平，并提高治疗性动物模型的长期存活率。

结论

我们的发现表明，这种新型融合蛋白疫苗能够通过改善 TC-1 荷瘤小鼠的 CD8 + T 细胞来诱导治疗效果和免疫原性；因此，这种疫苗可能会被用于针对 HPV 和肿瘤免疫疗法的研究。