十一标志物十色流式细胞术评估 T 细胞急性淋巴细胞白血病微小残留病的排除法。
Eleven-marker 10-color flow cytometric assessment of measurable residual disease for T-cell acute lymphoblastic leukemia using an approach of exclusion.
机构信息
Hematopathology Laboratory, ACTREC, Tata Memorial Center, HBNI University, Navi Mumbai, India.
Department of Pediatric Oncology, Tata Memorial Center, Tata Memorial Hospital, Parel, Mumbai, India.
出版信息
Cytometry B Clin Cytom. 2021 Jul;100(4):421-433. doi: 10.1002/cyto.b.21939. Epub 2020 Aug 19.
UNLABELLED
Measurable/minimal residual disease (MRD) status has been suggested as a powerful indicator of clinical-outcome in T-cell lymphoblastic leukemia/lymphoma (T-ALL). Multicolor flow cytometric (MFC)-based T-ALL MRD reports are limited and traditionally based on the utilization of markers-of-immaturity like TdT and CD99. Moreover, studies demonstrating the multicolor flow cytometric (MFC) approach for the assessment of T-ALL MRD are sparse. Herein, we describe an 11-marker, 10-color MFC-based T-ALL MRD method using an "approach of exclusion."
METHODS
The study included 269 childhood T-ALL patients treated with a modified-MCP841 protocol. An 11-marker, 10-color MFC-based MRD was performed in bone marrow (BM) samples at the end-of-induction (EOI) and end-of-consolidation (EOC) time-points using Kaluza-version-1.3 software.
RESULTS
We studied EOI-MRD in 269 and EOC-MRD in 105 childhood T-ALL patients. EOI-MRD was detectable in 125 (46.5%) samples (median, 0.3%; range, 0.0007-66.3%), and EOC-MRD was detectable in 34/105 (32.4%) samples (median, 0.055%; range, 0.0008-27.6%). Leukemia-associated immunophenotypes (LAIPs) found useful for MRD assessment were dual-negative CD4/CD8 (40.9%), dual-positive CD4/CD8 (23.3%) and only CD4 or CD8 expression (35.8%); dim/subset/dim-negative surface-CD3 (39%), dim/subset/dim-negative/negative CD5 (28.3%), dim/dim-negative/negative/heterogeneous CD45 (44.7%) and co-expression of CD5/CD56 (7.5%). EOI-MRD-positive status was found to be the most-relevant independent factor in the prediction of inferior relapse-free and overall survival.
CONCLUSION
We described an 11-marker 10-color MFC-based highly sensitive MRD assay in T-ALL using an approach of exclusion. The addition of CD4 and CD8 to the pan-T-cell markers in a 10-color assay is highly useful in T-ALL MRD assessment and extends its applicability to almost all T-ALL patients.
未标记
微量残留病（MRD）状态已被认为是 T 细胞淋巴母细胞白血病/淋巴瘤（T-ALL）临床结果的有力指标。多色流式细胞术（MFC）为基础的 T-ALL MRD 报告有限，传统上基于不成熟标志物如 TdT 和 CD99 的利用。此外，证明用于评估 T-ALL MRD 的多色流式细胞术（MFC）方法的研究很少。在此，我们描述了一种基于排除法的 11 标志物 10 色 MFC 基于 T-ALL MRD 方法。
方法
该研究纳入了 269 例接受改良 MCP841 方案治疗的儿童 T-ALL 患者。在诱导结束（EOI）和巩固结束（EOC）时，使用 Kaluza-version-1.3 软件，对骨髓（BM）样本进行基于 11 标志物 10 色 MFC 的 MRD 检测。
结果
我们研究了 269 例 EOI-MRD 和 105 例 EOC-MRD 儿童 T-ALL 患者。125 例（46.5%）样本可检测到 EOI-MRD（中位数为 0.3%；范围为 0.0007-66.3%），34/105 例（32.4%）样本可检测到 EOC-MRD（中位数为 0.055%；范围为 0.0008-27.6%）。用于 MRD 评估的白血病相关免疫表型（LAIPs）发现双阴性 CD4/CD8（40.9%）、双阳性 CD4/CD8（23.3%）和仅 CD4 或 CD8 表达（35.8%）、亚群/亚群/亚群阴性表面-CD3（39%）、亚群/亚群/亚群阴性/阴性 CD5（28.3%）、亚群/亚群/亚群阴性/阴性/异质 CD45（44.7%）和 CD5/CD56 的共表达（7.5%）对 MRD 评估非常有用。EOI-MRD 阳性状态被发现是预测复发无事件生存和总体生存的最相关独立因素。
结论
我们描述了一种基于排除法的 11 标志物 10 色 MFC 高度敏感的 T-ALL MRD 检测方法。在 10 色检测中，将 CD4 和 CD8 添加到泛 T 细胞标志物中，在 T-ALL MRD 评估中非常有用，并将其适用性扩展到几乎所有 T-ALL 患者。
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