多参数流式细胞术识别与急性髓系白血病微小残留病具有类似表型的再生细胞

doi:10.3969/j.issn.1004-583X.2024.08.007

摘要/Abstract

摘要：

目的探讨如何利用多参数流式细胞术(MFC)鉴别与急性髓系白血病(AML)微小残留病(MRD)具有类似表型的正常细胞。方法回顾性分析2020年3月-2022年4月在我院利用MFC检测的AML患者MRD骨髓标本157例次,识别容易被误认为是MRD的正常细胞。结果 AML患者治疗后再生的骨髓样本中会出现如下易误判为MRD的正常细胞群:CD117^dimCD56⁺CD7⁺CD45^str自然杀伤(NK)细胞、CD19^dimCD56⁺CD7⁺CD45^str NK细胞、CD300e⁺HLA-DR⁺CD14^partCD64^part非经典单核细胞;这些细胞群占骨髓有核细胞比例的中位值分别是0.084%(范围:0~0.6200%)、0%(范围:0~0.2134%)、0.1549%(范围:0~2.0940%)。结论 MFC检测AML患者MRD过程中,应避免将治疗后骨髓再生的正常细胞,误判为残留病。

关键词: 急性白血病, 多参数流式细胞术, 微小残留病, 正常细胞, 再生

Abstract:

Objective To discriminate normal cells with similar phenotypes to minimal residual disease (MRD) in acute myeloid leukemia (AML) by multi-parameter flow cytometry (MFC). Methods A total of 157 consecutive bone marrow aspirates for routine MRD between March 2020 to April 2022 were retrospectively analyzed. Normal cells misrecognized as MRD were identified. Results The following normal cell populations were easily misrecognized as MRD in regenerated bone marrow samples of AML patients after treatment: CD117^dimCD56⁺CD7⁺CD45^strnatural killer (NK) cells, CD19^dimCD56⁺CD7⁺CD45^str natural killer (NK) cells, and CD300e⁺HLA-DR⁺CD14^partCD64^part non-classical monocytes. The median proportion of these cell populations in the bone marrow nucleated cells was 0.084% (range, 0-0.6200%), 0% (range, 0-0.2134%), and 0.1549%(range, 0-2.0940%), respectively. Conclusion It is necessary to avoid a misdiagnosis of normal cells regenerated by bone marrow as MRD by MFC in AML patients after treatment.

Key words: acute leukemia, multi-parameter flow cytometry, minimal residual disease, normal cells, regeneration

中图分类号:

R733.7

范桥珍, 康蕊, 张婷. 多参数流式细胞术识别与急性髓系白血病微小残留病具有类似表型的再生细胞[J]. 临床荟萃, 2024, 39(8): 716-727.

Fan Qiaozhen, Kang Rui, Zhang Ting. Multi-parameter flow cytometry identifies regenerating cells with phenotypes similar to minimal residual disease in acute myeloid leukemia[J]. Clinical Focus, 2024, 39(8): 716-727.

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链接本文: https://huicui.hebmu.edu.cn/CN/10.3969/j.issn.1004-583X.2024.08.007

https://huicui.hebmu.edu.cn/CN/Y2024/V39/I8/716

图/表 5

表1 AML患者MRD检测的抗体及克隆号

Tab.1 Antibodies and clone numbers for MRD detection in patients with acute myeloid leukemia

Tube-1	Tube-2	Tube-3	Tube-4
CD7 FITC/M-T701	CD14 FITC/MφP9	CD71 FITC/L01.1	CD61 FITC/RUU-PL7F12
CD117 PE/104D2	CD64 PE/10.1	CD235a PE/GA-R2(HIR2)	CD41a PE/HIP8
CD34 Percp-cy5.5/8G12	CD34 Percp-cy5.5/8G12	CD34 Percp-cy5.5/8G12	CD34 Percp-cy5.5/8G12
CD19 PE-CY7/SJ25C1
CD11b APC/D12	CD300e APC/UP-H2	CD105 APC/266	CD42b APC/HIP1
HLA-DR APC-H7/L243	HLA-DR APC-H7/L243	CD117 APC-H7/2B8	CD117 APC-H7/2B8
CD56 BV421/HCD56
CD45 V500-C/2D1	CD45 V500-C/2D1	CD45 V500-C/2D1	CD45 V500-C/2D1

图1 AML MRD与治疗后正常增生的CD117dim NK细胞,CD19dim NK细胞比较

Fig.1 Comparison of antigen expression between MRD in AML with the normal proliferating CD117dim NK cells and CD19dim NK cells after treatment

表2 再生NK细胞在不同患者骨髓中比例(%)

Tab.2 Proportion of regenerated NK cells in BMs of different patients(%)

组别	例数	CD117^dimNK占比	CD19^dimNK占比
化疗组	70	0.1100(0.0000~0.1855)	0.0000(0.0000~0.0000)
移植组	87	0.0442(0.0361~0.0645)	0.0000(0.0000~0.0000)
$t$ 值		0.1704	14.59
$P$ 值		0.8649	<0.0001
组别	例数	CD117^dimNK占比	CD19^dimNK占比
MRD⁺组	14	0.0415(0.0360~0.0460)	0.0000(0.0000~0.0000)
MRD^-组	143	0.1100(0.0000~0.1855)	0.0000(0.0000~0.0000)
$t$ 值		2.4120	0.4816
$P$ 值		0.0170	0.6307

表2 再生NK细胞在不同患者骨髓中比例(%)

Tab.2 Proportion of regenerated NK cells in BMs of different patients(%)

组别	例数	CD117^dimNK占比	CD19^dimNK占比
化疗组	70	0.1100(0.0000~0.1855)	0.0000(0.0000~0.0000)
移植组	87	0.0442(0.0361~0.0645)	0.0000(0.0000~0.0000)
$t$ 值		0.1704	14.59
$P$ 值		0.8649	<0.0001
组别	例数	CD117^dimNK占比	CD19^dimNK占比
MRD⁺组	14	0.0415(0.0360~0.0460)	0.0000(0.0000~0.0000)
MRD^-组	143	0.1100(0.0000~0.1855)	0.0000(0.0000~0.0000)
$t$ 值		2.4120	0.4816
$P$ 值		0.0170	0.6307

图2 AML MRD与治疗后正常增生的非经典单核细胞比较

Fig.2 Comparison of antigen expression between MRD in AML with normal proliferating non-classical monocytes

表3 非经典单核细胞在不同患者骨髓中比例(%)

Tab.3 Proportion of non-classical monocytes in BMs of different patients(%)

组别	例数	非经典单核细胞占比
化疗组	70	0.1800(0.1006~1.7661)
移植组	87	0.1346(0.0286~0.3108)
$t$ 值		1.7550
$P$ 值		0.0812
组别	例数	非经典单核细胞占比
MRD⁺组	14	0.0357(0.0000~0.2094)
MRD^-组	143	0.1570(0.0369~0.3312)
$t$ 值		0.5861
$P$ 值		0.5587

表3 非经典单核细胞在不同患者骨髓中比例(%)

Tab.3 Proportion of non-classical monocytes in BMs of different patients(%)

组别	例数	非经典单核细胞占比
化疗组	70	0.1800(0.1006~1.7661)
移植组	87	0.1346(0.0286~0.3108)
$t$ 值		1.7550
$P$ 值		0.0812
组别	例数	非经典单核细胞占比
MRD⁺组	14	0.0357(0.0000~0.2094)
MRD^-组	143	0.1570(0.0369~0.3312)
$t$ 值		0.5861
$P$ 值		0.5587

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