基于肝癌特异性γδTCR转染细胞系筛选γδT细胞识别抗原肽的实验研究

Screening of Antigen Peptides Recognized byγδT Cells Based on Hepatoma SpecificγδTCR Transfected Cell Lines

目的:利用肝癌特异性γδTCR转染细胞系筛选γδT细胞识别抗原肽。方法:收集肝癌患者外周血中的单个核细胞,测定其中的γ9和δ2的CDR3序列,根据序列分析获得优势的CDR3序列,嵌入到长的γ9δ2序列。分别构建到PREP7和PREP9载体中,转染J.RT-T3.5细胞,经过抗生素筛选,获得肝癌特异性γδTCR转染细胞。以此细胞为探针,筛选噬菌体十二肽库,获得γδT细胞识别抗原肽。结果:成功构建肝癌特异性γδTCR转染细胞,并筛选了可能的γδT细胞识别的肝癌特异性抗原肽。结论:本实验研究为筛选γδT细胞识别的肝癌特异性抗原奠定基础。

Objective To screen antigen peptides recognized byγδT cells by hepatoma specificγδTCR transfected cell lines.Methods The peripheral blood mononuclear cells from patients with hepatocellular carcinoma were collected and CDR3 sequences ofγ9 andδ2 were determined.According to the sequence analysis,the dominant CDR3 sequence was ob⁃tained and inserted into the full-lengthγ9 andδ2 sequence.They were constructed into PREP7 and PREP9 vectors,and transfected into J.RT-T3.5 cells.After antibiotic screening,liver cancer-specificγδTCR transfected cells were obtained.Using this cell as a probe,we screened the phage dodecapeptide library to obtainγδT cell recognition antigen peptides.Re⁃sults Hepatoma-specificγδTCR transfected cells were successfully constructed,and hepatoma-specific antigen peptides recognized byγδT cells were screened.Conclusion This study laid a foundation for screening hepatoma-specific antigen recognized byγδT cells.