T细胞精细亚群免疫监测在B淋巴细胞增殖性疾病中的应用  

Application of T-lymphocyte detailed subsets immunosurveillance in B-lymphoproliferative disorder

作　　者：田瑞丰 董晓雅 卢菲[1] 李国盛 Tian Ruifeng;Dong Xiaoya;Lu fei;Li Guosheng(Department of Hematology,Qilu Hospital of Shandong University,Jinan 250012,China)

机构地区：[1]山东大学齐鲁医院血液科,济南250012

出　　处：《中华检验医学杂志》2024年第12期1426-1434,共9页Chinese Journal of Laboratory Medicine

基　　金：山东省医学会资助项目(YXH2022ZX03215);山东省医学会青年人才托举工程(2023_LC_0336)。

摘　　要：目的分析T细胞精细亚群在B淋巴细胞增殖性疾病(B-LPD)患者和健康人群中的细胞占比差异及细胞免疫监测特征。方法观察性研究。收集山东大学齐鲁医院2022年5月14日至2024年1月19日间诊断收治累及骨髓和外周血的B-LPD患者194例[男122例、女72例、年龄(64±20)岁], 其中包括慢性淋巴细胞白血病(CLL)76例、弥漫大B细胞淋巴瘤(DLBCL)23例、滤泡淋巴瘤(FL)17例、毛细胞白血病(HCL)12例、淋巴浆细胞淋巴瘤/巨球蛋白血症(LPL/WM)10例、套细胞淋巴瘤(MCL)26例、边缘区淋巴瘤(MZL)20例, 以及CD5-B型淋巴细胞增殖性疾病未分类(B-LPD-U)10例。同时收集本院健康管理中心2023年11月18日采集的健康对照组(HC)45名[男22名、女23名、年龄(62±17)岁]。利用流式细胞术对T细胞亚群进行精细分群, 根据圈门策略, 按照CD45RA和CD62L的表达情况将CD4^(+)和CD8^(+)T细胞分别划分初始T细胞(TN, CD45RA^(+)CD62L^(+))、中央记忆T细胞(TCM, CD45RA-CD62L^(+))、效应记忆T细胞(TEM, CD45RA-CD62L^(-))、终末分化效应记忆细胞(TEMRA, CD45RA^(+)CD62L^(-));将调节性T细胞(Treg, CD3^(+)CD4^(+)CD127^(low) CD25^(high))划分为CD45RA^(+)的初始Treg(nTreg)和CD45RA-的记忆Treg(mTreg), 以及HLA-DR^(+)的活化Treg(aTreg)。采用Kruskal-Wallis单因素方差分析比较HC组与各组B-LPD患者的T细胞亚群差异变化。结果 B-LPD各组的CD3^(+)T细胞在淋巴细胞的比例(CCL组13.15%、DLBCL组41.75%、FL组30.52%、HCL组34.24%、LPL/WM组40.58%、MCL组20.67%、MZL组26.36%、CD5-B-LPD-U组17.49%)均小于HC组(64.85%)。B-LPD各组的CD4^(+)T细胞在CD3^(+)T细胞的比例(CCL组46.63%、DLBCL组40.76%、FL组42.77%、HCL组43.81%、LPL/WM组43.02%、MCL组45.58%、MZL组43.95%、CD5-B-LPD-U组46.91%)均低于HC组(54.61%), 差异有统计学意义。结论 B-LPD相关疾病影响T细胞免疫功能, CD4^(+)TEM增多、CD8^(+)TEMRA减少以及aTreg的增多提示B-LPD抑制CD8^(+)T细胞免疫杀伤, 激活Treg细胞免疫�Objective:To investigate the difference in the proportions of T-lymphocyte subsets and the immunological monitoring profiles between patients with B-lymphoproliferative disorder(B-LPD)and healthy individuals.Methods:This clinical observational study involved 194 B-LPD patients[122 males,72 females,average age(64±20)]treated in Qilu Hospital of Shandong University from May 14th,2022 to January 19th,2024.The patient cohort included 76 cases of chronic lymphocytic leukemia(CLL),23 cases of diffuse large B-cell lymphoma(DLBCL),17 cases of follicular lymphoma(FL),12 cases of hair-cell leukemia(HCL),10 cases of lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia(LPL/WM),26 cases of mantle cell lymphoma(MCL),20 cases of marginal zone lymphoma(MZL),and 10 cases of unclassified CD5-B lymphocytic proliferative disease unclassified(B-LPD-U).At the same time,45 health control(HC)samples[22 males,23 females,aged 62(±17)]from the medical examination center of our hospital in November 18th,2023 were collected.The T cell subsets were meticulously analyzed using flow cytometry,including CD4^(+)naive T cell[CD4^(+)TN(CD3^(+)CD4^(+)CD45RA^(+)CD62L^(+))],CD4^(+)central memory T cell[CD4^(+)TCM(CD3^(+)CD4^(+)CD45RA-CD62L^(+))],CD4^(+)terminally differentiated effector memory cells expressing CD45RA[CD4^(+)TEMRA(CD3^(+)CD4^(+)CD45RA^(+)CD62L^(-))],CD4^(+)effector memory T cell[CD4^(+)TEM(CD3^(+)CD4^(+)CD45RA-CD62L^(-))],CD8^(+)TN(CD3^(+)CD8^(+)CD45RA^(+)CD62L^(+)),CD8^(+)TCM(CD3^(+)CD8^(+)CD45RA-CD62L^(+)),CD8^(+)TEMRA(CD3^(+)CD8^(+)CD45RA^(+)CD62L^(-)),CD8^(+)TEM(CD3^(+)CD8^(+)CD45RA-CD62L^(-)),naive regulatory T cell[nTreg(CD3^(+)CD4^(+)CD127^(dim+)CD25^(+)CD45RA^(+))],memory Treg[mTreg(CD3^(+)CD4^(+)CD127^(dim+)CD25^(+)CD45RA-)]and activated Treg[aTreg(CD3^(+)CD4^(+)CD127 ^(dim+)CD25^(+)HLA-DR^(+))].The difference of T cell subsets data were compared by Mann-Whitney test.Result:The proportionof CD3^(+)T cells within lymphocyte in B-LPD groups(CCL group 13.15%,DLBCL group 41.75%,FL group 30.52%,HCL group 34.24%,LPL/WM group 4

关 键 词：淋巴组织增殖性疾病 B细胞淋巴增殖性疾病 CD4^(+)T细胞 CD8^(+)T细胞 调节性T细胞 流式细胞术 

分 类 号：R733[医药卫生—肿瘤]