Hebei Yanda Lu Daopei Hospital, Department of Pathology and Laboratory Medicine, Langfang, China
Beijing Lu Daopei Hospital, Department of Pathology and Laboratory Medicine, Beijing, China
Turk J Haematol. 2023 Feb 28;40(1):18-27. doi: 10.4274/tjh.galenos.2022.2022.0365. Epub 2023 Jan 31.
In minimal residual disease (MRD) analysis after allogeneic hematopoietic stem cell transplantation (allo-HSCT), abnormal immunophenotyping is commonly considered as evidence of a secondary recurrence or complications, leading to overtreatment. We aimed to confirm whether such phenotypic abnormality might originate from donors using multicolor flow cytometry (MFC).
The MRD of bone marrow specimens of 3395 patients who had received allo-HSCT were analyzed using the conventional two-tube, eight-color MFC panel. The frequencies of abnormal immunophenotypes were also evaluated in three groups of patients without malignancies.
The frequency of new abnormal polymorphisms was 0.088% (3/3395) among patients who received allo-HSCT. The abnormal cells seen in three patients in complete remission were Fcγ receptor IIIB (FcγRIIIB) gene deletion (CD16- neutrophils), CD2-CD159a-CD159c natural killer (NK) cells, and monoclonal B lymphocytosis (MBL), respectively. In addition, abnormal T-cells (CD4CD8) were detected in one donor before allo-HSCT. Identical abnormalities were found in the peripheral blood of the corresponding donors of the three patients via MFC. Among the individuals without malignancies, the incidence of FcγRIIIB deletion was 0.2% (11/5256), that of NK cells with the absence of CD2 and single-positive CD159c was 0.05% (1/2000), that of monoclonal CD4/CD8 double-positive T-cells was 0.05% (1/2000), and that of MBL was 1.3% (14/1100). The frequency of NK cells with the absence of CD2 was 1.3% (1/79) and with CD8 was 14% (11/79) in NK cell lymphoma. The following abnormalities could be identified by the two-tube, eight-color MFC panel: cκ/cλ/CD19/CD5/CD20/ CD38/CD45/CD56 (adding CD10 and CD34 as the ninth and tenth colors) and CD16CD56/CD5/CD3/CD7/CD4/CD8/CD2/CD45 (adding CD117 as the ninth color).
Abnormalities in recipients of allo-HSCT detected by MRD analysis may originate from their donors. Screening of donor specimens with a suitable two-tube, eight- to ten-color MFC panel may be a promising method for minimizing misdiagnoses.
在异基因造血干细胞移植（allo-HSCT）后微小残留病（MRD）分析中，异常免疫表型通常被认为是二次复发或并发症的证据，导致过度治疗。我们旨在通过多色流式细胞术（MFC）确认这种表型异常是否可能源自供者。
使用常规的两管八色 MFC 面板分析了 3395 例接受 allo-HSCT 的患者的骨髓标本中的 MRD。还评估了三组无恶性肿瘤患者的异常免疫表型频率。
allo-HSCT 患者新出现异常多态性的频率为 0.088%（3/3395）。在完全缓解的 3 名患者中，异常细胞分别为 Fcγ 受体 IIIB（FcγRIIIB）基因缺失（CD16-中性粒细胞）、CD2-CD159a-CD159c 自然杀伤（NK）细胞和单克隆 B 淋巴细胞增多症（MBL）。此外，在 allo-HSCT 前，一名供者中还检测到异常 T 细胞（CD4+CD8+）。通过 MFC 在 3 名患者的相应供者的外周血中均发现了相同的异常。在无恶性肿瘤的个体中，FcγRIIIB 缺失的发生率为 0.2%（11/5256），无 CD2 和单阳性 CD159c 的 NK 细胞的发生率为 0.05%（1/2000），单阳性 CD4/CD8 双阳性 T 细胞的发生率为 0.05%（1/2000），MBL 的发生率为 1.3%（14/1100）。NK 细胞淋巴瘤中无 CD2 的 NK 细胞的发生率为 1.3%（1/79），CD8 的发生率为 14%（11/79）。两管八色 MFC 面板可识别以下异常：cκ/cλ/CD19/CD5/CD20/CD38/CD45/CD56（添加 CD10 和 CD34 作为第九和第十色）和 CD16+CD56/CD5/CD3/CD7/CD4/CD8/CD2/CD45（添加 CD117 作为第九色）。
MRD 分析中接受 allo-HSCT 的受者检测到的异常可能源自供者。使用合适的两管、八到十色 MFC 面板对供者标本进行筛查可能是减少误诊的一种有前途的方法。
