CTP-BCR-ABL抗原肽负载树突状细胞体外致敏T淋巴细胞靶向杀伤CML细胞  

Cytotoxic T Lymphocytes Sensitized in Vitro by Dendritic Cells Loaded with CTP-BCR-ABL Antigen Peptide Can Target and Kill CML Cells

作　　者：王旭 曾大川 郑忍忍 黄峥兰[1] 高淼[2] WANG Xu;ZENG Dachuan;ZHENG Renren;HUANG Zhenglan;GAO Miao(Key Laboratory of Clinical Laboratory Diagnostics Ministry of Education,School of Laboratory Medicine,Chongqing Medical University,Chongqing 400016,China;Department of Laboratory Medicine,the First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China)

机构地区：[1]重庆医科大学检验医学院,临床检验诊断学教育部重点实验室,重庆400016 [2]重庆医科大学附属第一医院检验科,重庆400016

出　　处：《中国细胞生物学学报》2021年第2期353-362,共10页Chinese Journal of Cell Biology

基　　金：国家自然科学基金青年基金(批准号:81802074);重庆市基础科学与前沿技术研究项目(批准号:cstc2018jcyjAX0726)资助的课题。

摘　　要：BCR-ABL为慢性髓细胞白血病特异胞质抗原,为良好的免疫治疗靶标。该研究选择BCR-ABL融合位点的两段抗原肽SSKALQRPV(SS)、GFKQSSKAL(GF)为靶点,与胞质转导肽融合表达,负载小鼠骨髓源性树突状细胞。在胞质转导肽介导下,SS、GF短肽进入树突状细胞并定位于内质网,具备了被树突状细胞识别为内源性抗原并以MHC I类分子递呈的条件。在体外培养中,用致敏的树突状细胞刺激脾脏CD8^(+)T淋巴细胞,获得针对CML的细胞毒性T淋巴细胞,同时检测该细胞毒性T淋巴细胞体外抗CML的效应。结果证实,胞质转导肽介导的GF抗原短肽负载的树突状细胞能够诱导CD8^(+)T淋巴细胞增殖活化并产生针对CML的细胞毒性杀伤效应。因此,GF抗原肽有望作为CML免疫治疗的靶点。该研究为鉴定出靶向CML细胞的T淋巴细胞表面的特异TCR序列准备了条件,进而为后续制备靶向CML的TCR-T细胞奠定了基础。BCR-ABL is a specific cytoplasmic antigen of chronic myeloid leukemia,which is suitable to be a target for immunotherapy.In this study,the two antigen peptides,SSKALQRPV(SS)and GFKQSSKAL(GF),at the BCR-ABL fusion site were selected as targets.SS and GF were respectively fused with CTP(cytoplasmic transduction peptide).The fusion peptides were used to sensitize bone marrow-derived dendritic cells of Balb/c mouse.Under the mediation of cytoplasmic transduction peptide,CTP-SS and CTP-GF peptides smoothly entered the cytoplasm of dendritic cells and located in the endoplasmic reticulum,thus having the basic conditions to be recognized as endogenous antigens and be presented to the cell surface by MHC I molecules.In vitro culture,the spleen CD8^(+)T lymphocytes were stimulated by dendritic cells sensitized with CTP-SS or CTP-GF to obtain cytotoxic T lymphocytes against chronic myelogenous leukemia cells.Then,the effect of cytotoxic T lymphocytes against CML cells in vitro was tested.The results confirmed that dendritic cells loaded with CTP-GF antigen peptide could induce the proliferation and activation of CD8^(+)T lymphocytes and produce cytotoxic killing effects against CML cells.Therefore,the GF antigen peptide at the BCR-ABL fusion site is expected to be a well target for CML immunotherapy.This study prepares the conditions for identifying specific TCR sequences on the surface of T lymphocytes that target CML cells,and then lays the foundation for our subsequent preparation of TCR-T cells that target CML.

关 键 词：树突状细胞 细胞毒性T淋巴细胞 BCR-ABL 慢性髓细胞白血病 胞质转导肽 

分 类 号：R733.72[医药卫生—肿瘤]