Improved Recognition of Hematogones From Precursor B-Lymphoblastic Leukemia by a Single Tube Flow Cytometric Analysis

Don, Michelle; Lim, Washington; Lo, Amanda; Cox, Brian; Huang, Qin; Kitahara, Sumire; Lopategui, Jean; Alkan, Serhan

doi:10.1093/ajcp/aqaa007

Cited by 5 publications

(3 citation statements)

References 34 publications

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“…When evaluating normal B cell maturation, we confirmed a dim to positive expression of CD58 by early hematogones, with a gradual reduction of this level of expression during differentiation to the mature B cell stage, with a significant difference between hematogones I or II and B mature lymphocytes ( p = 0.0040 and p = 0.0009, respectively, Figure S4A), as previously observed (Chen et al, 2001; Don et al, 2020; Lee et al, 2005).…”

Section: Resultssupporting

confidence: 89%

Immunophenotypic portrait of leukemia‐associated‐phenotype markers in B acute lymphoblastic leukemia

Boris,

Theron,

Montagnon

et al. 2023

Cytometry Part B Clinical

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BackgroundMultiparametric flow cytometry (MFC) is an essential diagnostic tool in B acute lymphoblastic leukemia (B ALL) to determine the B‐lineage affiliation of the blast population and to define their complete immunophenotypic profile. Most MFC strategies used in routine laboratories include leukemia‐associated phenotype (LAP) markers, whose expression profiles can be difficult to interpret. The aim of our study was to reach a better understanding of 7 LAP markers' landscape in B ALL: CD9, CD21, CD66c, CD58, CD81, CD123, and NG2.MethodsUsing a 10‐color MFC approach, we evaluated the level of expression of 7 LAP markers including CD9, CD21, CD66c, CD58, CD81, CD123, and NG2, at the surface of normal peripheral blood leukocytes (n = 10 healthy donors), of normal precursor B regenerative cells (n = 40 uninvolved bone marrow samples) and of lymphoblasts (n = 100 peripheral blood samples or bone marrow samples from B ALL patients at diagnosis). The expression profile of B lymphoblasts was analyzed according the presence or absence of recurrent cytogenetic aberrations. The prognostic value of the 7 LAP markers was examined using Maxstat R algorithm.ResultsIn order to help the interpretation of the MFC data in routine laboratories, we first determined internal positive and negative populations among normal leukocytes for each of the seven evaluated LAP markers. Second, their profile of expression was evaluated in normal B cell differentiation in comparison with B lymphoblasts to establish a synopsis of their expression in normal hematogones. We then evaluated the frequency of expression of these LAP markers at the surface of B lymphoblasts at diagnosis of B ALL. CD9 was expressed in 60% of the cases, CD21 in only 3% of the cases, CD58 in 96% of the cases, CD66c in 45% of the cases, CD81 in 97% of the cases, CD123 in 72% of the cases, and NG2 in only 2% of the cases. We confirmed the interest of the CD81/CD58 MFI expression ratio as a way to discriminate hematogones from lymphoblasts. We observed a significant lower expression of CD9 and of CD81 at the surface of B lymphoblasts with a t(9;22)(BCR‐ABL) in comparison with B lymphoblasts without any recurrent cytogenetic alteration (p = 0.0317 and p = 0.0011, respectively) and with B lymphoblasts harboring other cytogenetic recurrent abnormalities (p = 0.0032 and p < 0.0001, respectively). B lymphoblasts with t(1;19) at diagnosis significantly overexpressed CD81 when compared with B lymphoblasts with other recurrent cytogenetic abnormalities or without any recurrent alteration (p = 0.0001). An overexpression of CD58 was also observed in the cases harboring this abnormal cytogenetic event, when compared with B lymphoblasts with other recurrent cytogenetic abnormalities (p = 0.030), or without any recurrent alteration (p = 0.0002). In addition, a high expression of CD123, of CD58 and of CD81 was associated with a favorable prognosis in our cohort of pediatric and young adult B ALL patients. We finally built a risk score based on the expression of these 3 LAP markers, this scoring approach being able to split these patients into a high‐risk group (17%) and a better outcome group (83%, p < 0.0001).ConclusionThe complexity of the phenotypic signature of lymphoblasts at diagnosis of B ALL is illustrated by the variability in the expression of LAP antigens. Knowledge of the expression levels of these markers in normal leukocytes and during normal B differentiation is crucial for an optimal interpretation of diagnostic cytometry results and serves as a basis for the biological follow‐up of B ALL.

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Section: Resultssupporting

confidence: 89%

Immunophenotypic portrait of leukemia‐associated‐phenotype markers in B acute lymphoblastic leukemia

Boris,

Theron,

Montagnon

et al. 2023

Cytometry Part B Clinical

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“…Approximately 75% of B-ALL cases express CD38, with a dimmer intensity than that expressed on HGs. The applicability of the underexpression of CD38 in B-ALL as a discriminating marker has been demonstrated in various studies examining its utility in MRD testing [ 11 ].…”

Section: Discussionmentioning

confidence: 99%

Unusual B-Lymphoid Blastic Crisis as Initial Presentation of Chronic Myeloid Leukemia Imposes Diagnostic Challenges

Momen

Baysal

Sait

et al. 2022

Case Reports in Hematology

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Chronic myeloid leukemia (CML) is a clonal hematopoietic stem cell disorder, characterized by reciprocal translocation t(9,22) (q34; q11), leading to increased myeloid proliferation. Most cases are diagnosed in the chronic phase (CP). However, a minority of cases can be present in the blastic phase (BP). In most patients with CML-BP, the blasts have a myeloid phenotype, however, in 20–30% of cases, the blasts have a lymphoid phenotype, mostly a B-cell phenotype. It is challenging to differentiate CML B-lymphoblastic phase (CML-BLP) from Ph + primary B-acute lymphoblastic leukemia (B-ALL) especially when the CML-BLP is the initial presentation of the disease, which is uncommon. We report here an unusual case of CML-BLP as an initial presentation of the disease without typical CML morphological findings. This case demonstrates diagnostic challenges and emphasizes the importance of an integrated approach using morphology, multiparametric flow cytometry, cytogenetic studies, and molecular studies to render an accurate diagnosis.

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“…The inclusion of CD22 allows gating of B cells in patients where CD19 may be lost or masked by targeted therapy (Cherian et al, 2018). CD58 is commonly used in other B‐ALL MRD panels (Coustan‐Smith et al, 2002; Keeney et al, 2018), and has been shown to be informative as an MRD marker (Shaver et al, 2015) and in differentiating haematogones from B‐ALL blasts (Don et al, 2020). Cost is also an important consideration for diagnostic laboratories, and the antibody panel we present is considerably less expensive than the EuroFlow panel.…”

Section: Discussionmentioning

confidence: 99%

Correlation between a 10‐color flow cytometric measurable residual disease (MRD) analysis and molecular MRD in adult B‐acute lymphoblastic leukemia

Singh¹,

Gorniak²,

Grigoriadis

et al. 2021

Cytometry Part B Clinical

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Background Measurable residual disease (MRD) monitoring in acute lymphoblastic leukemia (ALL) is an important predictive factor for patient outcome and treatment intensification. Molecular monitoring, particularly with quantitative polymerase chain reaction (qPCR) to measure immunoglobin heavy or kappa chain (Ig) or T‐cell receptor (TCR) gene rearrangements, offers high sensitivity but accessibility is limited by expertise, cost, and turnaround time. Flow cytometric assays are cheaper and more widely available, and sensitivity is improved with multi‐parameter flow cytometry at eight or more colors. Methods We developed a 10‐color single tube flow cytometry assay. Samples were subject to bulk ammonium chloride lysis to maximize cell yields with a target of 1 × 106 events. Once normal maturation patterns were established, patient samples were analyzed in parallel to standard molecular monitoring. Results Flow cytometry was performed on 114 samples. An informative immunophenotype was identifiable in all 22 patients who had a diagnostic sample. MRD analysis was performed on 87 samples. The median lower limits of detection and quantification were 0.004% (range 0.0005%–0.028%) and 0.01% (range 0.001%–0.07%) respectively. Sixty‐five samples had concurrent molecular MRD testing, with good correlation (r = 0.83, p < 0.001). Results were concordant in 52 samples, and discordant in 13 samples, including one case where impending relapse was detected by flow cytometry but not Ig/TCR qPCR. Conclusions Our 10‐color flow cytometric MRD assay provided adequate sensitivity and good correlation with molecular assays. This technique offers rapid and affordable testing in B‐ALL patients, including cases where a suitable molecular assay cannot be developed or has reduced sensitivity.

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Improved Recognition of Hematogones From Precursor B-Lymphoblastic Leukemia by a Single Tube Flow Cytometric Analysis

Cited by 5 publications

References 34 publications

Immunophenotypic portrait of leukemia‐associated‐phenotype markers in B acute lymphoblastic leukemia

Immunophenotypic portrait of leukemia‐associated‐phenotype markers in B acute lymphoblastic leukemia

Unusual B-Lymphoid Blastic Crisis as Initial Presentation of Chronic Myeloid Leukemia Imposes Diagnostic Challenges

Correlation between a 10‐color flow cytometric measurable residual disease (MRD) analysis and molecular MRD in adult B‐acute lymphoblastic leukemia

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