HPV18E7抗原HLA-A2限制性CTL表位的初步筛选

Preliminary screening of HLA-A2-restricted CTL epitope derived from human papillomavirus 18E7 antigen

目的:初步筛选人乳头瘤病毒18型E7抗原的HLA-A2限制性细胞毒性T淋巴细胞(CytotoxicTlymphocyte,CTL)表位,为表位免疫原性鉴定提供靶肽。方法:采用超基序、多项式和量化基序方案相结合的方法,对靶抗原HPV18E7的HLA-A2限制性细胞毒性T淋巴细胞表位进行预测,并应用固相合成法合成多肽,经RP-HPLC纯化及纯度分析,用质谱进行定性鉴定。结果:分别预测出5条、1条和6条九肽表位,综合三种预测方案确定其中1条为候选合成表位,合成肽的纯度大于95%,经质谱分析合成肽的分子量测定值与理论值相符。结论:超基序、多项式和量化基序方案联合应用提高预测效率,避免了在研究HPV18E7抗原表位时盲目合成多肽;所合成的多肽为高纯度靶肽,为后继的抗原表位的免疫原性的鉴定奠定了实验基础。

Objective：To screen the HLA-A2-restricted CTL epitopes of human papilloma virus 18 E7 antigen, and to identify the predicted HLA-A2-restricted CTL epitope candidate from HPV18E7 for immunogenicity identification. Methods：The CTL epitopes of HPV18E7 antigen were predicted by HLA-A2-binding peptide supermotif prediction method combined with the polynomial and quantitative motif method.The peptide was synthesized with solid phase strategies and identified with mass spectrometry, the purity of the peptide was detected by reverse-phase high performance liquid chromatography （RP-HPLC）. Results： Five,one and six CIL epitopes were respectively predicted, among which one peptide was the candidate to be synthesized. The peptide was beyond 95% in purity and the measured value of molecular weight conformed to its theoretical value. Conclusions： The conbination of supermotif with the polynomial and quantitative motif method can improve the prediction efficiency and avoid synthesizing peptides aimlessly about HPV18E7 antigen. The synthesized peptide is the target peptide with high purity,the study provides basis for immunogenicity identification in future.