Wilms’tumor 1(WT1)抗原HLA-A11限制性T细胞表位鉴定及特异性TCR筛选  

Identification of HLA-A11 restricted T cell epitope of Wilms’tumor 1(WT1)antigen and isolation of WT1-specific TCR

作　　者：蒋敏 孙文桥 卢丹 贺娟华 王杰 谭曙光[3] 高福 Min Jiang;Wenqiao Sun;Dan Lu;Juanhua He;Jie Wang;Shuguang Tan;George F.Gao(Savaid Medical School,University of Chinese Academy of Sciences,Beijing 100049,China;Institutes of Physical Science and Information Technology,Anhui University,Hefei 230601,China;CAS Key Laboratory of Pathogenic Microbiology and Immunology,Institute of Microbiology,Chinese Academy of Sciences,Beijing 100101,China;College of Life Sciences,Jiangxi Science and Technology Normal University,Nanchang 330013,China)

机构地区：[1]中国科学院大学存济医学院,北京100049 [2]安徽大学物质科学与信息技术研究院,合肥230601 [3]中国科学院微生物研究所,中国科学院病原微生物与免疫学重点实验室,北京100101 [4]江西科技师范大学生命科学学院,南昌330013

出　　处：《科学通报》2022年第17期1945-1957,共13页Chinese Science Bulletin

基　　金：国家重点研发计划(2021YFC2301400);国家自然科学基金(92169208)资助。

摘　　要：Wilms’tumor 1(WT1)基因编码的WT1蛋白是在多种血液系统恶性肿瘤以及实体肿瘤中过表达的肿瘤相关抗原(TAA),靶向WT1表位的T细胞受体工程T细胞(TCR-T)疗法在预防和治疗血液瘤的研究中显示出潜在的临床应用价值.本研究通过生物信息学预测了WT1蛋白HLA-A11限制性T细胞表位多肽,并通过免疫HLA-A*11:01转基因小鼠鉴定出了一条CD8^(+)T细胞表位WT1_(414-423)(序列:FSCRWPSCQK).通过WT1_(414-423)/HLA-A11四聚体染色和单细胞分选,获得了WT1_(414-423)表位特异性TCR,命名为9E TCR.细胞水平的结合实验以及蛋白水平的亲和力检测表明,9E TCR可以特异性结合WT1_(414-423)/HLA-A11.进一步通过携带9E TCR的慢病毒感染来自健康个体外周血的原代T细胞制备了9E TCR-T细胞,其与呈递WT1_(414-423)多肽的PANC-1靶细胞孵育后可诱导9E TCR-T细胞特异性分泌IFN-γ.WT1_(414-423)多肽表位的鉴定为A11遗传背景人群的肿瘤免疫治疗提供了新的靶标,9E TCR的发现为抗肿瘤TCR-T细胞治疗药物开发提供了重要基础.The WT1 protein,encoded by the Wilms’tumor 1(WT1)gene,is a tumor-associated antigen(TAA)and is overexpressed in a variety of hematopoietic malignant tumors and solid tumors.WT1 protein functions as a potent transcriptional regulator that regulates cell proliferation,cell death,and differentiation.WT1 protein is essential for the development of kidney,uterus,and gonad.Upregulated expression of WT1 on tumor cells is associated with aggressiveness and poor prognosis in multiple cancers,such as breast cancer,germ cell tumors,prostate cancer,and soft tissue sarcoma.In recent years,the development of tumor immunotherapy,especially adoptive T cell therapy(ACT),has substantially improved the prognosis of patients with metastatic cancer.Five CAR-T(chimeric antigen receptors-T)therapeutic agents have been commercially approved for clinical treatment of B-cell lymphoma or multiple myeloma,whereas CAR-T therapy is less effective for solid tumors.The T-cell receptor engineered T cell(TCR-T)therapy can target intracellular antigens of tumor cells presented by human leukocyte antigen(HLA)molecules through TCR,and has shown a tumorsuppressive effect on hematological and solid tumors in several clinical trials.A study of TCR-T cell therapy in acute myeloid leukemia(AML)patients found that adoptive infusion of TCR-T cells targeting the HLA-A2-restricted WT1epitope into patients can effectively prevent AML relapse after bone marrow transplantation.HLA-A11 is one of the most common HLA class I genotypes around the world,with a phenotypic frequency of about 20%–30%in the Chinese population.No HLA-A11-restricted T cell epitope has been reported for the WT1 antigen,which restricted the development of immune therapeutic agents for HLA-A11 patients.In this study,HLA-A11 restricted T cell epitopes of WT1 protein were in silico predicted and a CD8^(+)T cell epitope WT1_(414-423)(FSCRWPSCQK)was identified by immunizing HLA-A*11:01 transgenic mice.A WT1_(414-423)epitope-specific murine TCR,named 9E TCR,was obtained by WT1_(414-423)/HLA-A

关 键 词：WT1 HLA-A11 T细胞表位 TCR 免疫疗法 

分 类 号：R730.51[医药卫生—肿瘤]