中国人群优势HLA-A分子限制性甲胎蛋白T细胞表位谱研究  

T cell epitope spectrum of alpha-fetoprotein restricted by dominant HLA-A molecules in Chinese population

作　　者：杨松 朱苏月 吴怡 孙万亮 刘中 沈传来[2] 鲁正 Yang Song;Zhu Suyue;Wu Yi;Sun Wanliang;Liu Zhong;Shen Chuanlai;Lu Zheng(Department of Hepatobiliary and Pancreatic Surgery,the First Affiliated Hospital of Bengbu Medical College,Bengbu 233000,China;Department of Microbiology and Immunology,Medical School of Southeast University,Nanjing 210009,China)

机构地区：[1]蚌埠医科大学第一附属医院肝胆胰外科,蚌埠233000 [2]东南大学医学院病原生物学与免疫学系,南京210009

出　　处：《中华微生物学和免疫学杂志》2024年第9期752-761,共10页Chinese Journal of Microbiology and Immunology

基　　金：国家自然科学基金面上项目(82272402)。

摘　　要：目的研究被13种中国人群优势人类白细胞抗原A(human leukocyte antigen A,HLA-A)分子限制的甲胎蛋白(alpha-fetoprotein,AFP)T细胞表位谱。方法利用多种T细胞表位预测数据库,虚拟预测AFP抗原被13种HLA-A分子限制的候选表位肽。采用多肽-肝癌患者外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)共刺激试验(多肽-PBMCs试验)和胞内细胞因子染色验证候选表位肽的免疫原性。利用表达特定HLA-A分子的12种HMy2.CIR细胞株进行HLA-A分子的多肽竞争结合试验,分析每种HLA-A分子与相应表位肽的亲和力和交叉限制性。结果通过对67例AFP阳性肝细胞癌患者进行多肽-PBMCs共刺激试验,从42种候选表位肽中获得20种能刺激患者CD8^(+)T细胞反应的阳性表位肽。HLA-A分子的多肽竞争结合试验显示,与特定HLA-A分子呈高、中、低亲和力的表位肽分别有22、13和6种候选表位肽,另有10种无亲和力;20种经多肽-PBMCs共刺激实验验证的阳性表位肽均呈现与相应HLA-A分子的高或中亲和力结合,且多数肽能与多种HLA-A分子交叉结合。结论通过肝细胞癌患者的T细胞功能性试验获得20种被中国人群优势HLA-A分子限制的AFP特异性CD8^(+)T细胞表位肽,并初步明确了这些表位肽与对应HLA-A分子的交叉结合能力。本研究为设计中国人群普适性的AFP特异性T细胞检测系统和肝癌多肽疫苗等提供了较为系统的基础数据。ObjectiveTo investigate T cell epitope spectrum of alpha fetoprotein(AFP)restricted by 13 dominant human leukocyte antigen A(HLA-A)molecules in Chinese population.MethodsAFP T cell epitope candidates presented by 13 HLA-A molecules were in silico predicted using multiple epitope prediction algorithms.Then,the candidate epitope peptides were co-cultured in vitro with fresh peripheral blood mononuclear cells(PBMCs)from hepatocellular carcinoma(HCC)patients,followed by intracellular cytokine staining(ICS)and flow cytometry to verify the immunogenicity of the candidate epitope peptides.Peptide competition binding assay of HLA-A molecules were performed using 12 HMy2.CIR cell lines expressing the indicated HLA-A molecules to analyze the affinity and cross-restriction of candidate epitope peptides with HLA-A molecules.ResultsPBMCs from 67 AFP+HCC patients were co-cultured in vitro with 42 candidate T cell epitope peptides,and the result showed that 20 epitope peptides stimulated CD8+T cell responses(named as positive epitope peptides).Peptide competitive binding assay revealed 22 candidate epitope peptides with high affinity,13 with inter affinity,six with low affinity,and 10 without affinity with indicated HLA-A molecules,respectively.The 20 positive epitope peptides presented high or inter affinity with corresponding HLA-A molecules and most of them displayed cross-binding properties with several HLA-A molecules.ConclusionsTwenty AFP-specific CD8+T cell epitope peptides restricted by the predominant HLA-A molecules in Chinese population are obtained by the HCC patients-derived T-cell functional experiments,and the cross-binding ability of these epitope peptides to the corresponding HLA-A molecules is preliminary identified.These results provide systematic and fundamental data for the design of AFP-specific T cell detection systems and T cell epitope-based vaccines universal for the Chinese population.

关 键 词：肝细胞癌 甲胎蛋白 T细胞表位 人类白细胞抗原A 抗原特异性T细胞 

分 类 号：R735.7[医药卫生—肿瘤]