Multi-parameter flow cytometry identifies regenerating cells with phenotypes similar to minimal residual disease in acute myeloid leukemia

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

Abstract

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

CLC Number:

R733.7

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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URL: https://huicui.hebmu.edu.cn/EN/10.3969/j.issn.1004-583X.2024.08.007

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

Figures/Tables 5

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

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

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

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

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

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

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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