Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu, 610065, Sichuan, People's Republic of China.
Bio-resource Research and Utilization Joint Key Laboratory of Sichuan and Chongqing, Chongqing, People's Republic of China.
Virol J. 2019 May 28;16(1):72. doi: 10.1186/s12985-019-1168-y.
Human papillomavirus (HPV) E6 and E7 oncoproteins play a crucial role in HPV-related diseases, such as cervical cancer, and can be used as ideal targets for therapeutic vaccines. Human leukocyte antigen (HLA) participates in the immune response to block HPV infection and invasion by its target/recognition function. HPV-33 and HPV-58 are highly prevalent among Chinese women. Therefore, it is of great significance to study the E6 and E7 region-specific gene polymorphisms of HPV-33 and HPV-58 in Southwest China and to identify ideal epitopes for vaccine design. Both HPV-33 and HPV-58 belong to α-9 genus HPV and are highly homologous, so their correlations are included in our research.
To study the E6 and E7 variations and polymorphisms of HPV-33 and HPV-58 in Southwest China, we collected samples, extracted and sequenced DNA, and identified variants. Nucleotide sequences were translated into amino acids by Mega 6.0 software. The physical/chemical properties, amino acid-conserved sequences and secondary structure of protein sequences were analysed by the Protparam server, ConSurf server and PSIPRED software. The T and B cell epitopes of the E6/E7 reference and variant sequences in HPV-33 and HPV-58 were predicted by the Immune Epitope Database (IEDB) analysis server and the ABCpred server, respectively.
Five and seven optimal HLA-I restricted T cell epitopes were selected from HPV-33 and HPV-58 E6, respectively, and these optimal epitopes are mainly located in EVYDFAFADLTVVYREGN of HPV-33 E6 and SEVYDFVFADLRIVYRDGNPF of HPV-58 E6. Six optimal HLA-I-restricted T cell epitopes were selected from HPV-33 and HPV-58 E7, and these epitopes are mainly located in RTIQQLLMGTVNIV of HPV-33 E7 and RTLQQLLMGTCTIV of HPV-58 E7.
HPV-33/HPV-58 E6/E7 gene polymorphisms and T/B cell epitopes of their reference and variant sequences were studied, and candidate epitopes were selected by bioinformatics techniques for therapeutic vaccine design for people in Southwest China. This study was the first to investigate the correlation of epitopes between HPV-33 and HPV-58. After experimental validation, these selected epitopes will be employed to induce a wide range of immune responses in heterogeneous HLA populations.
人乳头瘤病毒（HPV）E6 和 E7 癌蛋白在 HPV 相关疾病（如宫颈癌）中发挥着关键作用，可作为治疗性疫苗的理想靶点。人类白细胞抗原（HLA）通过其靶标/识别功能参与免疫反应，以阻断 HPV 感染和入侵。HPV-33 和 HPV-58 在我国女性中高度流行。因此，研究中国西南地区 HPV-33 和 HPV-58 的 E6 和 E7 区域特异性基因突变，并鉴定疫苗设计的理想表位具有重要意义。HPV-33 和 HPV-58 均属于α-9 属 HPV，高度同源，因此它们的相关性包含在我们的研究中。
为研究中国西南地区 HPV-33 和 HPV-58 的 E6 和 E7 变异和多态性，我们采集样本，提取并测序 DNA，并鉴定变异。Mega 6.0 软件将核苷酸序列翻译成氨基酸。Protparam 服务器、ConSurf 服务器和 PSIPRED 软件分析蛋白质序列的理化性质、氨基酸保守序列和二级结构。免疫表位数据库（IEDB）分析服务器和 ABCpred 服务器分别预测 HPV-33 和 HPV-58 的 E6/E7 参考和变异序列的 T 细胞和 B 细胞表位。
从 HPV-33 和 HPV-58 的 E6 中分别选择了 5 个和 7 个最佳 HLA-I 限制性 T 细胞表位，这些最佳表位主要位于 HPV-33 E6 的 EVYDFAFADLTVVYREGN 和 HPV-58 E6 的 SEVYDFVFADLRIVYRDGNPF 中。从 HPV-33 和 HPV-58 的 E7 中分别选择了 6 个最佳 HLA-I 限制性 T 细胞表位，这些表位主要位于 HPV-33 E7 的 RTIQQLLMGTVNIV 和 HPV-58 E7 的 RTLQQLLMGTCTIV 中。
通过生物信息学技术研究 HPV-33/HPV-58 E6/E7 基因突变及其参考和变异序列的 T/B 细胞表位，选择候选表位用于设计针对中国西南地区人群的治疗性疫苗。本研究首次探讨了 HPV-33 和 HPV-58 之间表位的相关性。经过实验验证，这些选择的表位将被用于诱导异质 HLA 人群中广泛的免疫反应。
