Brief communication: Distribution of bone marrow cell subsets and hemodilution in patients with acute leukemia

Géner, G.; Espasa, Andrea; Raya, Minerva; Vergara, Sara; Juncà, Jordi; Sorigué, Marc

doi:10.1111/ijlh.13243

Cited by 6 publications

(5 citation statements)

References 9 publications

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“…Altogether, this points out the need for new cellular markers and approaches for more accurate estimation of BM hemodilution. In this regard, some authors have used paired PB and BM samples and defined specific formulas to estimate BM hemodilution based on both absolute and/or relative cell counts as well as hemoglobin levels [34,41,42]. However, such approaches are not easy to implement in routine laboratory diagnostics, pointing out the need for more simple and user-friendly approaches to estimate BM hemodilution in individual patients.…”

Section: Discussionmentioning

confidence: 99%

B-Cell Regeneration Profile and Minimal Residual Disease Status in Bone Marrow of Treated Multiple Myeloma Patients

Pontes

Flores‐Montero

Sanoja-Flores

et al. 2021

Cancers

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B-cell regeneration during therapy has been considered as a strong prognostic factor in multiple myeloma (MM). However, the effects of therapy and hemodilution in bone marrow (BM) B-cell recovery have not been systematically evaluated during follow-up. MM (n = 177) and adult (≥50y) healthy donor (HD; n = 14) BM samples were studied by next-generation flow (NGF) to simultaneously assess measurable residual disease (MRD) and residual normal B-cell populations. BM hemodilution was detected in 41 out of 177 (23%) patient samples, leading to lower total B-cell, B-cell precursor (BCP) and normal plasma cell (nPC) counts. Among MM BM, decreased percentages (vs. HD) of BCP, transitional/naïve B-cell (TBC/NBC) and nPC populations were observed at diagnosis. BM BCP increased after induction therapy, whereas TBC/NBC counts remained abnormally low. At day+100 postautologous stem cell transplantation, a greater increase in BCP with recovered TBC/NBC cell numbers but persistently low memory B-cell and nPC counts were found. At the end of therapy, complete response (CR) BM samples showed higher CD19− nPC counts vs. non-CR specimens. MRD positivity was associated with higher BCP and nPC percentages. Hemodilution showed a negative impact on BM B-cell distribution. Different BM B-cell regeneration profiles are present in MM at diagnosis and after therapy with no significant association with patient outcome.

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Section: Discussionmentioning

confidence: 99%

B-Cell Regeneration Profile and Minimal Residual Disease Status in Bone Marrow of Treated Multiple Myeloma Patients

Pontes

Flores‐Montero

Sanoja-Flores

et al. 2021

Cancers

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“…We assessed hemodilution as immature granulocyte count (SS high /CD10‐neg) over neutrophils (SS high /CD10‐pos) (IGRA/N), adapting a previously reported method (Gener et al, 2020). Greater values correspond with higher quality samples.…”

Section: Methodsmentioning

confidence: 99%

Flow cytometry to detect bone marrow involvement by follicular lymphoma

Aren

Marcé

Jurado

et al. 2022

Cytometry Part B Clinical

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Background High‐quality data on bone marrow involvement (BMI) assessed by flow cytometry (FC) in follicular lymphoma (FL) is lacking. Aims We set up a prospective protocol with a 10‐color tube and acquisition of 500.000 leukocytes on a Nav flow cytometer for evaluation of BMI in FL by FC. Materials and methods FC was compared with a combination of histopathology and IGH gene rearrangement, which were considered the gold standard. We also compared BMI by FC with PET. Results Fifty‐two patients were included (median 67 years, 54% female). BMI by FC was seen in 35 (67%), with a median involvement of 1.2% (interquartile range: 0.3%–7%) of leukocytes. Comparison with the gold standard revealed two false negatives and two false positives (potentially true involvement undetected by the gold standard). BMI by PET was seen in 14/46 (30%). Immunophenotype of FL in the bone marrow was highly heterogeneous. The most common phenotypic abnormality was dim expression of CD19 (>0.5 log loss in 30% of patients). CD10 was negative in 13 (37%) and incompletely positive (overlap with the negative population) in a further 8 (28%) while entirely positive only in 14 (48%). Other abnormalities (loss of CD20, gain or loss of CD79b, expression of CD43, and substantial loss of CD45) were rare. Computational analysis by means of FlowSOM confirmed the heterogeneous phenotype, with FL from different patients clustering in unrelated metaclusters. Conclusion BMI by FL was frequent and immunophenotype was heterogeneous. However, this protocol enabled detection of FL in bone marrow in the vast majority of patients with bone marrow involvement by the gold standard.

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“…All patients included in the study provided written informed consents for biological studies and have been treated following local recommendations for clinical trials or routine clinical practice at the Seràgnoli Institute, IRCCS Azienda Ospedaliero-Universitaria of Bologna, Italy. An initial cohort of 78 Multiple Myeloma (MM) and 18 Acute Lymphoblastic Leukemia (ALL) patients (104 and 34 samples, respectively) was used to set-up the experimental plan to evaluate the cellular immunophenotype by flow cytometry (FC) of the most representative BM cell types and to create the matrix to compare diverse hemodilution strategies, previously described by other groups ( 16 – 20 ). ALL patients are represented by Philadelphia (Ph)-negative B-ALL or T-ALL patients, for whom minimal residual disease (MRD) is measured molecularly by Next Generation Sequencing (NGS).…”

Section: Methodsmentioning

confidence: 99%

“…and Julie Pont herself had already studied both the correlation between Pont’s IG/N and the formula defined by Holdrinet et al. (HI) ( 18 , 20 ), defining an R of 0.8 (p-value <0.001) in healthy individuals, we decided to use IG/N ratio as reference for our considerations. We excluded the possibility to employ the HI as a reference, even though it is considered as the gold standard to assess hemodilution by most authors, since it requires both a parallel PB sampling and a parallel characterization and count of leukocytes and erythrocytes.…”

Section: Methodsmentioning

confidence: 99%

The ALLgorithMM: How to define the hemodilution of bone marrow samples in lymphoproliferative diseases

Vigliotta

Armuzzi²,

Barone³

et al. 2022

Front. Oncol.

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IntroductionMinimal residual disease (MRD) is commonly assessed in bone marrow (BM) aspirate. However, sample quality can impair the MRD measurement, leading to underestimated residual cells and to false negative results. To define a reliable and reproducible method for the assessment of BM hemodilution, several flow cytometry (FC) strategies for hemodilution evaluation have been compared.MethodsFor each BM sample, cells populations with a well-known distribution in BM and peripheral blood - e.g., mast cells (MC), immature (IG) and mature granulocytes (N) – have been studied by FC and quantified alongside the BM differential count.ResultsThe frequencies of cells’ populations were correlated to the IG/N ratio, highlighting a mild correlation with MCs and erythroblasts (R=0.25 and R=0.38 respectively, with p-value=0.0006 and 0.0000052), whereas no significant correlation was found with B or T-cells. The mild correlation between IG/N, erythroblasts and MCs supported the combined use of these parameters to evaluate BM hemodilution, hence the optimization of the ALLgorithMM. Once validated, the ALLgorithMM was employed to evaluate the dilution status of BM samples in the context of MRD assessment. Overall, we found that 32% of FC and 52% of Next Generation Sequencing (NGS) analyses were MRD negative in samples resulted hemodiluted (HD) or at least mildly hemodiluted (mHD).ConclusionsThe high frequency of MRD-negative results in both HD and mHD samples implies the presence of possible false negative MRD measurements, impairing the correct assessment of patients’ response to therapy and highlighs the importance to evaluate BM hemodilution.

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Brief communication: Distribution of bone marrow cell subsets and hemodilution in patients with acute leukemia

Cited by 6 publications

References 9 publications

B-Cell Regeneration Profile and Minimal Residual Disease Status in Bone Marrow of Treated Multiple Myeloma Patients

B-Cell Regeneration Profile and Minimal Residual Disease Status in Bone Marrow of Treated Multiple Myeloma Patients

Flow cytometry to detect bone marrow involvement by follicular lymphoma

The ALLgorithMM: How to define the hemodilution of bone marrow samples in lymphoproliferative diseases

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