嵌合抗原受体-T细胞免疫治疗中流式细胞术动态监测调节性-T淋巴细胞水平及其临床意义  

The level of regulatory T lymphocyte detected by flow cytometry in chimeric antigen receptor T cell immunotherapy and its clinical significance

作　　者：毛霞 程佳丽 李好 王春艳 刘松雅 何成 易淑娟 朱莉 Mao Xia;Cheng Jiali;Li Hao;Wang Chunyan;Liu Songya;He Cheng;Yi Shujuan;Zhu Li(Department of Hematology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]华中科技大学同济医学院附属同济医院血液内科,武汉430030

出　　处：《中华检验医学杂志》2022年第8期825-832,共8页Chinese Journal of Laboratory Medicine

基　　金：国家自然科学基金(81900187)。

摘　　要：目的探究接受嵌合抗原受体(CAR)-T细胞免疫治疗的B细胞淋巴瘤患者外周血调节性T淋巴细胞(Treg)水平的变化特点,以及Treg水平与最佳疗效及治疗反应的联系。方法回顾性研究2019年至2021年在武汉同济医院接受CD19/CD22 CAR-T细胞免疫治疗的23例复发/难治性B细胞恶性肿瘤患者的资料,依据Lugano修订版的淋巴瘤疗效评价标准将入组患者按照最佳疗效分为治疗后完全缓解(CR)组8例,部分缓解(PR)组7例及无效组(NR)8例。收集武汉同济医院同期16例未进行CAR-T细胞免疫治疗的B细胞淋巴瘤患者为对照组。在CAR-T细胞免疫治疗期间不同时间段,应用多色流式细胞术动态检测患者外周血中Treg占CD4^(+)T细胞比例(Treg/CD4^(+)T)、占淋巴细胞比例(Treg/Lym)、占白细胞比例(Treg/WBC)及Treg绝对数(Treg#),分析Treg水平随时间的变化趋势以及回输前后不同时间段内Treg占比及绝对数在不同最佳疗效患者组别中的差异。根据CAR-T细胞回输后1~15 d内Treg占比和绝对数中位数水平将患者分为低水平组11例及高水平组12例,比较各组间CAR-T拷贝数峰值、铁蛋白峰值、白细胞介素6(IL-6)峰值的统计学差异。统计学分析采用独立样本t检验,曼-惠特尼U检验、Cox-Stuart趋势存在性检验及单因素方差分析。结果接受CAR-T细胞免疫治疗的23例患者,清淋预处理后CAR-T细胞输注前Treg/CD4^(+)T及Treg/Lym均值分别为(20.42±7.96)%及(13.61±7.13)%,显著高于对照组Treg/CD4^(+)T均值[(7.33±3.61)%,t=5.893,P<0.001]及Treg/Lym均值[(1.91±0.90)%,t=6.53,P<0.001]。同期Treg#均值(1.81±1.52)/μl明显低于对照组均值[(13.66±9.89)/μl,t=4.261,P<0.001]。回输CAR-T细胞后,Treg/CD4^(+)T与Treg/Lym均有降低趋势(P<0.001),Treg/WBC有升高趋势(P=0.01)。最佳疗效为CR组的患者回输前Treg/CD4^(+)T(12.87±1.93)%、Treg/Lym(6.35±2.84)%、Treg/WBC(0.05±0.05)%均值均明显低于PR组[(29.68±5.49)%(P<0.01),(21.85±2.10)%(P<0.01),0.50±0Objective To investigate the characteristics of changes in peripheral blood regulatory T lymphocyte(Treg)levels in patients with B-cell lymphoma who received chimeric antigen receptor(CAR)-T cell immunotherapy,and the relationship between Treg levels and optimal efficacy and treatment response.Methods The data of 23 patients with relapsed/refractory B-cell malignancies who received CD19/CD22 CAR-T cell immunotherapy in Wuhan Tongji Hospital from 2019 to 2021 were retrospectively studied.The enrolled patients were divided into complete remission(CR)group(8 cases),partial remission(PR)group(7 cases)and no response(NR)group(8 cases)according to Lugano′s revised lymphoma efficacy evaluation criteria.A total of 16 patients with B-cell lymphoma who did not receive CAR-T cell immunotherapy during the same period in Wuhan Tongji Hospital were collected as the control group.In different periods during CAR-T cell immunotherapy,multicolor flow cytometry(MFC)was used to dynamically detect peripheral blood the proportion of Treg in CD4^(+)T cells(Treg/CD4^(+)T),the proportion of lymphocytes(Treg/Lym),the proportion of Treg in white blood cells(Treg/WBC),and the absolute number of Treg(Treg#).The trend of Treg levels over time,as well as the differences in Treg levels in patients with different prognosis groups in different periods were analyzed.According to the proportion of Treg and the median level of absolute number within 1 to 15 days after CAR-T cell infusion,the patients were divided into a low-level group with 11 cases and a high-level group with 12 cases.The statistical differences in the peak value of CAR-T copy,iron protein,and IL-6 were compared between various groups.Independent samples t test,Mann-Whitney U test,Cox-Stuart trend existence test and one-way analysis of variance was used in statistical analysis.Results In the 23 patients who received CAR-T cell immunotherapy,the mean values of Treg/CD4^(+)T and Treg/Lym before CAR-T cell infusion were(20.42±7.96)%and(13.61±7.13)%,respectively,which were signific

关 键 词：受体 抗原 T细胞 嵌合抗原受体T细胞 免疫治疗 调节性T淋巴细胞 流式细胞术 

分 类 号：R733.1[医药卫生—肿瘤]