Ai L L, Lai A L, Qin X H, Liu B C, Li J, Wang J X, Zhu P
State Key Laboratory of Experimental Hematology, National Clinical Research Center for Blood Diseases, Haihe Laboratory of Cell Ecosystem, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin 300020, China Tianjin Institutes of Health Science, Tianjin 301600, China.
State Key Laboratory of Coordination Chemistry, Chemistry and Biomedicine Innovation Center (ChemBIC), School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023, China.
Zhonghua Xue Ye Xue Za Zhi. 2023 Jul 14;44(7):543-549. doi: 10.3760/cma.j.issn.0253-2727.2023.07.003.
This study aimed to explore the application of interaction-dependent fucosyl-biotinylation (FucoID), a chemical biology-based proximity labeling technique, in capturing tumor antigen-specific T cells and its clinical value in chronic myelogenous leukemia (CML) . Flow cytometry and fluorescence microscopy were employed to evaluate the experimental parameters for FucoID in CML. Peripheral blood samples were obtained from 14 newly diagnosed CML patients in the chronic phase. These samples underwent flow cytometry-based sorting and were subsequently labeled with FucoID to facilitate the isolation of tumor cells and T cells, followed by the immunophenotypic identification of tumor antigen-specific T cells. Finally, the diagnostic and therapeutic potential of FucoID in CML was assessed. Initially, the experimental parameters for FucoID in CML were established. The proportion of CD3(+) T cells in patients was (8.96±6.47) %, exhibiting a marked decrease compared with that in healthy individuals at (38.89±22.62) %. The proportion of tumor-specific antigen-reactive T cells was (3.34±4.49) %, which demonstrated interpatient variability. In addition, the proportion of tumor-specific antigen-active T cells in CD4(+) T cells was (3.95±1.72) %, which was generally lower than the proportion in CD8(+) T cells at (5.68±2.18) %. Compared with those in tumor-specific antigen-nonreactive T cells, CCR7(-)CD45RA(-) effector memory T cells and CCR7(-)CD45RA(+) effector T cells were highly enriched in tumor-specific antigen-reactive T cells. Moreover, the intensity of tumor immune reactivity in patients exhibited a significant correlation with white blood cell count (WBC) and hemoglobin (HGB) levels in peripheral blood, while no such correlation was observed with other clinical baseline characteristics. The combination of FucoID and flow cytometry enables the rapid identification and isolation of tumor antigen-specific T cells in CML. The successful application of this method in CML and the implications of our findings suggest its potential clinical value in the field of hematologic malignancies.
本研究旨在探索基于化学生物学的邻近标记技术——相互作用依赖岩藻糖基生物素化（FucoID）在捕获肿瘤抗原特异性T细胞中的应用及其在慢性髓性白血病（CML）中的临床价值。采用流式细胞术和荧光显微镜评估CML中FucoID的实验参数。从14例新诊断的慢性期CML患者中获取外周血样本。这些样本进行基于流式细胞术的分选，随后用FucoID标记以促进肿瘤细胞和T细胞的分离，接着对肿瘤抗原特异性T细胞进行免疫表型鉴定。最后，评估FucoID在CML中的诊断和治疗潜力。最初，确定了CML中FucoID的实验参数。患者中CD3(+) T细胞的比例为(8.96±6.47)%，与健康个体的(38.89±22.62)%相比显著降低。肿瘤特异性抗原反应性T细胞的比例为(3.34±4.49)%，显示出患者间的差异。此外，CD4(+) T细胞中肿瘤特异性抗原活性T细胞的比例为(3.95±1.72)%，总体低于CD8(+) T细胞中的比例(5.68±2.18)%。与肿瘤特异性抗原无反应性T细胞相比，CCR7(-)CD45RA(-)效应记忆T细胞和CCR7(-)CD45RA(+)效应T细胞在肿瘤特异性抗原反应性T细胞中高度富集。此外，患者的肿瘤免疫反应强度与外周血白细胞计数（WBC）和血红蛋白（HGB）水平显著相关，而与其他临床基线特征未观察到这种相关性。FucoID与流式细胞术的结合能够快速鉴定和分离CML中的肿瘤抗原特异性T细胞。该方法在CML中的成功应用以及我们研究结果的意义表明其在血液系统恶性肿瘤领域具有潜在的临床价值。
