Wang Y Z, Lu J, Hao L, Chang Y, He L L, Huang X J, Liu Y R
Peking University Institute of Hematology, People's Hospital of Peking University, Beijing Key Laboratory of Hematopoietic Stem Cell Transplantation, Beijing 100044, China.
Zhonghua Xue Ye Xue Za Zhi. 2017 Apr 14;38(4):272-278. doi: 10.3760/cma.j.issn.0253-2727.2017.04.003.
To explore and compare 4-color and 8-color fluorescence antibody panels for detecting minimal residual disease of multiple myeloma patients after therapy. One 8-color antibody panel was built including CD38 and CD138 for the identification of plasma cells (PCs) , membrane antigen CD45, CD19, CD56 and CD117, cytoplasmic Kappa (cκ) and Lambda (cλ) light chain antigen. Six tubes of 4-color panels were built, among them, membrane antigen CD45/CD19, CD56/CD117, CD19/CD56 and light chains were analyzed combined by CD38/CD138 for PCs gate in the tubes M1-3 and tube C-(κ/λ), respectively; CD19 or CD45 and cκ/cλ light chains were detected in the tube MC1-(C)D38 for CD38/SSC identified PCs gate and tube MC2-(CD138) for CD138/SSC identified PCs gate separately. Twenty normal volunteer bone marrows and seventy-three specimens from multiple myeloma patients after complete remission were measured and analyzed. MRD positive samples were discriminated in 82.19% of the specimen evaluated through either abnormal plasma cells (aPCs) or clonal plasma cells (cPCs) by 8-color antibody panel. Among of them, consistency was 89.04%. The median percentage of cPCs was 0.105%. The lowest sensitivity of experiment was 0.004%. Percentage of PCs identified by CD38/ SSC gate was higher than that by CD38/CD138 (<0.001) and CD138/SSC gate (=0.001) apparently. The lowest MRD positive rate was found in tube C (65.75%) , which lower than 8-color panel obviously (=0.024) . The percentages of aPCs measured by tube M2-(56/117) were significantly lower than other tubes (=0.014) . MRD positive rate determined by cPCs was higher than that by aPCs both in the tube MC1-(C)D38 and tube MC2-(CD138), whose concordance rate were 68.49% and 79.45%, respectively. Compared with 8-color panel, tube MC2-(CD138) the best choice among six tubes of 4-color panels, which has the best sensitivity, accuracy and negative predicted value, higher positive predicted value and specificity. Tube M1-(45/19) was the second choice. CD38/CD138 are the best markers for gating PCs. Membrane antigen and cκ/cλ detected simultaneously is a better method for MM MRD detection and 8-color antibody panel is an ideal approach. Two tubes of 4-color antibody combination, M1-(45/19) and MC2-(CD138) are recommended in the 4-color panels.
探索并比较4色和8色荧光抗体组合用于检测多发性骨髓瘤患者治疗后的微小残留病。构建了一个8色抗体组合,包括用于识别浆细胞(PCs)的CD38和CD138、膜抗原CD45、CD19、CD56和CD117、细胞质κ(cκ)和λ(cλ)轻链抗原。构建了6管4色组合,其中,在管M1 - 3和管C -(κ/λ)中,分别通过CD38/CD138对PCs门控,联合分析膜抗原CD45/CD19、CD56/CD117、CD19/CD56和轻链;在管MC1 -(C)D38中针对CD38/SSC识别的PCs门控检测CD19或CD45以及cκ/cλ轻链,在管MC2 -(CD138)中针对CD138/SSC识别的PCs门控分别检测。对20份正常志愿者骨髓和73份多发性骨髓瘤患者完全缓解后的标本进行检测和分析。通过8色抗体组合,在评估的标本中,82.19%的标本通过异常浆细胞(aPCs)或克隆浆细胞(cPCs)鉴别出微小残留病阳性样本。其中,一致性为89.04%。cPCs的中位百分比为0.105%。实验的最低灵敏度为0.004%。CD38/SSC门控识别的PCs百分比明显高于CD38/CD138(<0.001)和CD138/SSC门控(=0.001)。在管C中发现最低的微小残留病阳性率(65.75%),明显低于8色组合(=0.024)。管M2 -(56/117)检测的aPCs百分比显著低于其他管(=0.014)。在管MC1 -(C)D38和管MC2 -(CD138)中,通过cPCs确定的微小残留病阳性率均高于通过aPCs确定的,其符合率分别为68.49%和79.45%。与8色组合相比,管MC2 -(CD138)是4色组合6管中最佳选择,具有最佳的灵敏度、准确性和阴性预测值,较高的阳性预测值和特异性。管M1 -(45/19)是第二选择。CD38/CD138是门控PCs的最佳标志物。同时检测膜抗原和cκ/cλ是多发性骨髓瘤微小残留病检测的较好方法,8色抗体组合是理想的方法。在4色组合中推荐两管4色抗体组合,即M1 -(45/19)和MC2 -(CD138)。