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人乳头瘤病毒(human papillomavirus,HPV)早期基因E7是致癌的关键基因,其表达在宫颈癌细胞癌变进程及维持癌细胞恶性表型方面发挥重要作用,已成为宫颈癌治疗的理想靶标.目前,基于HPV16 E7抗原细胞毒性T淋巴细胞(cytotoxic lymphocyte,CTL)表位设计多肽疫苗是抗宫颈癌治疗发展的重要方向,但天然CTL表位肽普遍存在体内半衰期短、激发CTL反应效果不佳等缺点.因此,本研究基于前期HPV16 E7抗原CTL表位鉴定的基础,结合多肽酶解实验结果,进行分子动力学模拟及结合自由能计算,初步筛选了3条表位模拟肽.人工合成相关待测表位肽,并利用T2细胞株测定各肽与HLA-A2分子的结合力.研究结果表明,3条表位模拟肽体外抗酶解能力较天然HPV16 E7抗原CTL表位肽均有提高,以(d)RAHYNIVTF表位模拟肽的效果最为明显.此外,(d)RAHYNIVTF表位模拟肽与HLA-A2分子的结合力也有所提高(荧光系数为2.06).以上结果表明,基于HPV16 E7抗原CTL表位模拟肽进行结构修饰有望为宫颈癌治疗性疫苗的设计奠定基础.
The early gene E7 of human papillomavirus (HPV) is a key gene oncogenic which plays an important role in the carcinogenesis of cervical cancer cells and maintaining the malignant phenotype of cancer cells, and has become an ideal target for the treatment of cervical cancer.At present, Peptide vaccines based on HPV16 E7 antigen cytotoxic lymphocyte (CTL) epitopes are important directions for the development of anti-cervical cancer therapy. However, the natural CTL epitope peptides have the shortcomings of short half-life in vivo and poor response to CTL stimulation Therefore, based on the identification of pre-HPV16 E7 antigen CTL epitopes, three epitope-mimotope peptides were screened by molecular dynamics simulation and free energy calculation based on the experimental results of proteolysis of peptides.Artificial synthesis-related test Epitope peptide and the binding ability of each peptide to HLA-A2 molecule was determined by using T2 cell line.The results showed that the three epitope-mimicking peptides had higher anti-enzymatic activity than the native CTL epitope peptide of HPV16 E7 antigen in (d) The RAHYNIVTF epitope peptide is most effective, and (d) the binding affinity of RAHYNIVTF epitope peptide to HLA-A2 is also increased (fluorescence coefficient 2.06). It shows that structural modification is expected to lay the foundation for the design of therapeutic cervical cancer vaccine antigen of HPV16 E7 CTL epitope peptide-based simulation.