scholarly journals Detection of Del(17p) in Hematological Malignancies By Imaging Flow Cytometry

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 9-10
Author(s):  
Wendy N. Erber ◽  
Henry Hui ◽  
Jason Stanley ◽  
Thomas Mincherton ◽  
Kathryn Clarke ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Plasma Cell ◽  
Quantitative Data ◽  
Hematological Malignancies ◽  
Plasma Cells ◽  
Digital Images ◽  
Flow Cytometer ◽  
Imaging Flow Cytometry ◽  
Fish Signal ◽  
Chromosomal Defects

Del(17p) in chronic lymphocytic leukemia (CLL) and plasma cell myeloma has a unique genomic profile leading to refractoriness to conventional therapies and poor overall survival. Detection is generally by fluorescence in situ hybridization (FISH) on a slide with analysis of up to 200 nuclei, not necessarily all of being neoplastic cells. The small cell sample analyzed, and high threshold for a positive result (>5% positive cells) makes FISH a low precision test. This means false negative results are inevitable, especially if del(17p) is only in a minor clone, potentially leading to suboptimal treatment. To address this, we developed an automated FISH method with analysis on an imaging flow cytometer, an instrument with the functionality of a standard flow cytometer which generates high-resolution digital images of each cell. By combining immunophenotyping with FISH, on whole cells in a single test, we can detect chromosomal defects in cells with a specific phenotype. Aims: Our aim was to determine the capability of this automated integrated immunophenotyping-FISH imaging flow cytometry method to detect del(17p) in CLL and myeloma, the two commonest hematological malignancies, and in which this genetic defect encodes poor prognosis. We hypothesized that it would be more sensitive and specific than standard FISH. Methods: Bone marrow and/or blood samples in EDTA anticoagulant from 19 cases of CLL or myeloma, at diagnosis or on therapy, were studied. After red cell lysis, cells were incubated with fluorochrome-conjugated antibodies to CD3, CD5, CD19, CD38, and CD138 antigens (fluorochromes: BV480, BV605, AF647). Following fixation, cell membranes were permeabilized and double-stranded DNA denatured (78oC for 5 mins). FISH probes to the centromere of chromosome 17 (CEP17, Spectrum Green) and 17p12 locus (Orange-Red) were added and hybridized for 24 hours at 37oC. Nuclei were stained with SYTOX AADvanced. Data for up to 200,000 cells was collected on the Amnis® ImageStream®XMk II imaging flow cytometer. Digital images (x60) and quantitative data derived from computational algorithms (IDEAS software) were used to assess FISH signals overlying the nuclei of CD5/CD19-positive CLL cells or CD38/CD138-positive plasma cells for each probe. Digital images and quantitative data were assessed for FISH signals within immunophenotyped cells. Results: Between 10,000 and 200,000 (mean 60,000) cells were analyzed per sample. The FISH signals were seen on the digital images and confirmed by quantitative mean channel fluorescence intensity of the probes. There were 12 CLL cases with one 17p FISH signal in the CD5/CD19-positive population, with the number of del(17p) CLL cells ranging from 2 - 35% (or 0.4 - 23% of all cells analyzed) (Fig 1). This amounted to between 270 and 35,441 cells in the analysed sample with del(17p). The lowest del(17p) burden was in a patient on cytoreductive therapy. All CLL cells had normal diploid spots for the control CEP17 probe, and the CD3/CD5-positive T cells had dual signals for both CEP17 and 17p12 probes. There were 5 myeloma cases with 1 FISH signal for 17p overlying the nucleus of the CD38/CD138-positive plasma cells (Fig 2). In these cases, 52-90% of cells (15,600-18,000 cells) had a plasma cell phenotype and 13-19% of these (or 2,340-2,964 CD38/CD138-positive cells) showed del(17p). This represented 1-5% of all cells in the sample. All gated plasma cells had 2 FISH spots for CEP17. Conclusion: This imaging flow cytometry method that integrates FISH with immunophenotyping could detect del(17p) in CLL and myeloma with a lowest limit of detection of 0.4% and 1% respectively. The high sensitivity was achieved as many thousands of cells were analyzed, and 17p was only assessed in gated cells with the phenotype of interest (i.e. CD5/CD19 or CD38/CD138). This method, that includes positive cell identification by phenotype, precludes the need for prior cell sorting or purification. Imaging flow cytometry for del(17p) by "immuno-flowFISH" brings a new dimension to FISH analysis at diagnosis and for disease monitoring. Its high precision and specificity will enable detection of del(17p), even when only present in minor sub-clones, for therapeutic decision making and prognostic stratification. This technique has a real place in clinical assessment of del(17p) in CLL and myeloma and could be applied for other significant chromosomal defects of therapeutic and prognostic significance. Figure 1 Disclosures Augustson: Roche: Other: Support of parent study and funding of editorial support. Cheah:Celgene, F. Hoffmann-La Roche, Abbvie, MSD: Research Funding; Celgene, F. Hoffmann-La Roche, MSD, Janssen, Gilead, Ascentage Pharma, Acerta, Loxo Oncology, TG therapeutics: Honoraria.

Identification of the side population associated with ATP-binding cassette transporters activity using imaging flow cytometry

2021 ◽  
Vol 67 (2) ◽  
pp. 137-143
Author(s):  
A.M. Gisina ◽  
Y.S. Kim ◽  
K.N. Yarygin ◽  
A.Yu. Lupatov
Keyword(s):  
Flow Cytometry ◽  
Side Population ◽  
Flow Cytometer ◽  
Atp Binding ◽  
Tumor Resistance ◽  
Side Population Cells ◽  
Imaging Flow Cytometry ◽  
Atp Binding Cassette Transporters ◽  
Resistance To Chemotherapy ◽  
Atp Binding Cassette

DyeCycle Violet efflux, caused by ATP-binding cassette transporters activity, was analyzed in human colorectal adenocarcinoma cell lines SW480, HT-29, Caco-2 by neans of FACSAria III flow cytometer and ImageStreamX Mk II imaging flow cytometer. Along with similarity of cytometry data obtained on the two instruments, the use of imaging flow cytometry made it possible to characterize the morphology of side population cells, as well as morphology of other cell populations differing in the degree of dye accumulation. The population of cells, which are smaller than the side population cells and practically do not take the dye, is of the special interest. Probably, this population may contribute to the tumor resistance to chemotherapy.

scholarly journals Increased circulating plasma cells detected by flow cytometry predicts poor prognosis in patients with plasma cell myeloma

2017 ◽  
Vol 94 (3) ◽  
pp. 493-499 ◽  
Author(s):  
Mi Hyun Bae ◽  
Chan-Jeoung Park ◽  
Bo Hyun Kim ◽  
Young-Uk Cho ◽  
Seongsoo Jang ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Plasma Cell ◽  
Poor Prognosis ◽  
Plasma Cells ◽  
Plasma Cell Myeloma

A Single-Tube Seven-Colour Flow Cytometry Assay for Detection of Minimal Residual Disease In Myeloma.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 1688-1688
Author(s):  
Soraya Wuilleme ◽  
Nelly Robillard ◽  
Steven Richebourg ◽  
Marion Eveillard ◽  
Laurence Lodé ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Plasma Cell ◽  
Plasma Cells ◽  
Residual Disease ◽  
Flow Cytometry Assay ◽  
Single Tube ◽  
Cell Identification ◽  
Colour Flow ◽  
Multiparameter Analysis ◽  
Minimal Residual

Abstract Abstract 1688 The eradication of minimal residual disease (MRD) in myeloma predicts for improved outcome. A number of different approaches to myeloma MRD detection are available; these vary widely in sensitivity and cost. Flow cytometric assessment of MRD may be preferable in practice because of lower cost and easier feasibility. Myeloma MRD flow cytometry requires at least three markers for plasma cell identification (CD38, CD138 and CD45) and combination of several additional markers to detect phenotypic abnormality including CD19, CD20, CD27, CD28, CD45, CD56 and CD117. Also, assessment of immunoglobulin light-chain restriction (cytoplasmic K and L) combined with myeloma-associated phenotypic plasma cell abnormalities, is very important. Four-tube four-colour flow cytometry combine markers CD38/CD138/CD45 with markers for plasma cell phenotypic abnormalities and clonality. Six –colour flow cytometry combines the same markers (markers for plasma cell identification) plus clonality markers; it potentially increases the sensitivity of the method through coincident multiparameter analysis. However, the single-tube six-colour flow cytometry, proposed by others studies, excludes the myeloma-associated phenotypic plasma cell abnormalities and consequently decreases specificity of the assay. We propose a new single-tube seven-colour flow cytometry, including plasma cell identification antigens, clonality markers and myeloma-associated phenotypic plasma cell abnormalities markers. In this new method, PCs are stained with antibodies: (i) CD38, CD138, CD45 used for identified plasma cells and percentage plasma cells to total leucocytes. (ii) CD19 and CD56+CD28 used to identify normal and abnormal plasma cells; and (iii) cy-IgK and cy-IgL, for confirm the plasma cells clonality. We analysed normal bone marrow provided from healthy individuals. Our results showed a presence myeloma-associated phenotypic plasma cell abnormalities at low levels in healthy individual. The monotypy studies confirm polyclonality of this normal plasma cells. Then we compared MRD assessement with single-six colour flow cytometry assay (plasma cells markers, clonality markers and exluding myeloma-associated phenotypic markers) and seven-colour flow cytometry assay (including myeloma-associated phenotypic markers). Six –colour flow cytometry has a better sensitivity and showed efficacy for quantification MRD in myeloma patients. However, the single-tube six-colour flow cytometry excluded the myeloma-associated phenotypic plasma cell abnormalities and in some cases the seven-colour flow cytometry will be more informative because it detected myeloma-asociated phenotypic marquers combined with clonality marquers. Finally, the single-tube seven colour flow cytometry assay provides reduction in antibody cost and increases sensitivity and specificity of the method through coincident multiparameter analysis. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Detection of Circulating Tumor Plasma Cells in Monoclonal Gammopathies: Methods, Pathogenic Role, and Clinical Implications

Cancers ◽  
2020 ◽  
Vol 12 (6) ◽  
pp. 1499
Author(s):  
Luzalba Sanoja-Flores ◽  
Juan Flores-Montero ◽  
Martín Pérez-Andrés ◽  
Noemí Puig ◽  
Alberto Orfao
Keyword(s):  
Solid Tumors ◽  
Tumor Cells ◽  
Plasma Cell ◽  
Hematological Malignancies ◽  
Plasma Cells ◽  
Low Frequency ◽  
Strong Impact ◽  
Monoclonal Gammopathies ◽  
Plasma Cell Neoplasms ◽  
Cancer Dissemination

Cancer dissemination and distant metastasis most frequently require the release of tumor cells into the blood circulation, both in solid tumors and most hematological malignancies, including plasma cell neoplasms. However, detection of blood circulating tumor cells in solid tumors and some hematological malignancies, such as the majority of mature/peripheral B-cell lymphomas and monoclonal gammopathies, has long been a challenge due to their very low frequency. In recent years, the availability of highly-sensitive and standardized methods for the detection of circulating tumor plasma cells (CTPC) in monoclonal gammopathies, e.g., next-generation flow cytometry (NGF), demonstrated the systematic presence of CTPC in blood in virtually every smoldering (SMM) and symptomatic multiple myeloma (MM) patient studied at diagnosis, and in the majority of patients with newly-diagnosed monoclonal gammopathies of undetermined significance (MGUS). These methods set the basis for further detailed characterization of CTPC vs. their bone marrow counterpart in monoclonal gammopathies, to investigate their role in the biology of the disease, and to confirm their strong impact on patient outcome when measured both at diagnosis and after initiating therapy. Here, we review the currently available techniques for the detection of CTPC, and determine their biological features, physiopathological role and clinical significance in patients diagnosed with distinct diagnostic categories of plasma cell neoplasms.

Cyclooxygenase-2 (COX-2) Is Frequently Expressed in Multiple Myeloma (MM) and Is Associated with Poor Outcome.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2450-2450
Author(s):  
Marco Ladetto ◽  
Sonia Vallet ◽  
Andreas Trojan ◽  
Maria Dell’Aquila ◽  
Luigia Monitillo ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Plasma Cell ◽  
Cell Separation ◽  
Plasma Cells ◽  
Mrna Level ◽  
Prognostic Indicator ◽  
Disease Stage ◽  
Plasma Cell Dyscrasias ◽  
Cox 2 ◽  
Ht 29

Abstract Introduction. A perturbed microenvironment with secretion of inflammatory cytokines is typical of MM. Prostaglandins (pg) are implicated in inflammation and angiogenesis and play a role in the pathogenesis of several solid malignancies. Expression of COX-2, the key enzyme of pg synthesis in inflamed tissues, is common in many of these cancers and plays a major role in their development. Moreover, it often acts as a poor prognostic indicator. Despite a large amount of data concerning COX-2 expression in solid tumors, few data are currently available in hematological malignancies. In MM there are several biological, epidemiological and clinical considerations suggesting a potential involvement of the pg pathway. Aim of this study is to verify the involvement of COX-2 in MM and to assess its prognostic role. Patients and methods. COX-2 expression has been assessed by western blotting (WB) as previously described (Du Bois RN, et al, Gastroenterology, 1996). Our positive control was the COX-2 positive cell line HT-29, while bone marrows (BM) from 15 healthy donors were our negative controls. We assessed a panel of 124 samples obtained by 113 patients with plasma cell dyscrasias. Sixteen samples belonged to subjects with MGUS, 80 to patients with MM at diagnosis, and 28 to patients with relapsed/refractory MM. In 11 patients, samples taken at different treatment phases were available. To confirm WB findings and to demonstrate that COX-2 expression occurs in malignant plasma cells immunohistochemistry (IC), and flow cytometry for COX-2 were also performed in 31 and four patients, respectively. Finally, COX-2 expression has been assessed in BM cells from four COX-2 positive patients following selection for the CD138 antigen using the Miltenyi cell separation system. COX-2 expression at the mRNA level has also been assessed by real time quantitative PCR. Results. A dilution test showed that our technique is sensitive enough to detect 2% HT-29 cells in a background of COX-2 negative cells. The 15 normal BM were COX-2 negative. In contrast, COX-2 expression was noticed in 12.5% of MGUS, 34.6% of MM at diagnosis and 56% of MM at relapse. COX-2 positivity at diagnosis and relapse was unrelated to disease stage, BM plasmacytosis, creatinine, Hb levels and ß2 microglobulin. COX-2 expression appeared to be of prognostic relevance: at diagnosis the median time to progression was 14 months in COX-2 positive and 40 months in COX-2 negative subjects (p<0.001). At relapse, of 14 patients showing COX-2 expression, 10 have already died of MM, and four are still alive. In contrast, of 11 COX-2 negative patients only one patient died while 10 are still alive (p<0.001). IC, cell separation and flow cytometry studies indicate that COX-2 expression is related to the malignant plasma cell population. COX-2 mRNA was overexpressed in patients showing increased COX-2 protein expression. Conclusions: a) COX-2 is frequently expressed in plasma cell dyscrasias; b) COX-2 expression is more frequent in advanced disease phases; c) COX-2 expression correlates to a worse outcome. Future studies are required to verify whether COX-2 might be clinically useful as a prognostic marker and/or therapeutic target.

Evaluation of a 5-Color Flow Cytometric Technique for Immunophenotyping and Quantitation of Plasma Cell Myeloma in Post-Therapy Bone Marrows.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 5036-5036
Author(s):  
Tove Isaacson ◽  
Andrzej Jakubowiak ◽  
Lloyd Stoolman ◽  
Usha Kota ◽  
William Finn ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Minimal Residual Disease ◽  
Plasma Cell ◽  
Plasma Cells ◽  
Residual Disease ◽  
Plasma Cell Myeloma ◽  
Color Flow ◽  
Flow Cytometric ◽  
Minimal Residual

Abstract Multiparameter flow cytometry is a useful tool for comprehensive immunophenotyping of plasma cell myeloma, and has been proposed as a sensitive method for the evaluation of minimal residual disease in patients following treatment. This study aimed to assess the value of flow cytometry in quantitation of residual disease, in comparison to routine morphologic examination of first-pull bone marrow aspirate smears, in myeloma patients post-therapy. Heparinized bone marrow aspirates were obtained from 27 treated patients with plasma cell myeloma. Cells were prepared for 5-color flow cytometric analysis within 24-hours of specimen draw. Surface membrane staining with anti-CD19, CD20, CD38, CD45, CD56, and CD138 was followed by ammonium chloride lysis of red cells. Fixed and permeabilized cells were analyzed for cytoplasmic light chains to confirm clonality. Data were acquired using an FC500 flow cytometer (Beckman-Coulter), analyzed with CXP software with plasma cells isolated based on bright CD38+ or CD138+ expression. A median of 97,639 cellular events (range 14,279 to 262,508) were collected per analysis. Flow cytometric enumeration of plasma cells was compared to 500-cell differential counts of Wright-Giemsa-stained first-pull aspirate smears from the same cases. The median plasma cell count as determined by flow cytometry was 0.5% (range 0–7.9%). The median plasma cell count estimated by morphologic review was 8.0% (range 0–84.4%). Flow cytometry underestimated the plasma cell content in all but one case. Clonal plasma cells expressed CD38 and CD138 in all cases; 87.5% (21/24) coexpressed CD56, 25% (6/24) coexpressed CD45, and 4.2% (1/24) coexpressed CD19. None was positive for CD20. Although detection of minimal residual disease after therapy for acute leukemia is routinely achieved by flow cytometric analysis, successful quantitation of minimal residual disease in treated myeloma patients using flow cytometry remains limited as it usually underestimates the plasma cell content of bone marrow samples compared to routine morphology of first-pull aspirates. We have observed that this holds true for both pre-treatment and post-treatment specimens. Causes for the discrepancy may include hemodilution of second-pull aspirates used for flow cytometry, fragility and loss of plasma cells during preparation for flow cytometry, and incomplete disaggregation of plasma cells from bone marrow spicules. With improved outcome of treatments, better and more reliable methods of detection of minimal residual disease are needed for optimal prognostic stratification. We are currently validating alternative methods, which may offer more sensitivity while at the same time allow more objectivity, for assessing the amount of minimal residual disease in myeloma patients.

Increased Circulating Plasma Cells Predicts Poor Overall Survival in Symptomatic Plasma Cell Myeloma Patients

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 4205-4205
Author(s):  
Mi Hyun Bae ◽  
Sang Hyuk Park ◽  
Chan-Jeoung Park ◽  
Bo Hyun Kim ◽  
Young-Uk Cho ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Overall Survival ◽  
Plasma Cell ◽  
Peripheral Blood ◽  
Plasma Cells ◽  
Survival Curve ◽  
Plasma Cell Myeloma ◽  
Newly Diagnosed ◽  
Kaplan Meier ◽  
Meier Survival Curve

Abstract Backgrounds Flow cytometry can rapidly determine immunophenotypes of neoplastic plasma cells (PCs) and quantify PCs in patients with plasma cell myeloma. Flow cytometric immunophenotyping and quantification of neoplastic plasma cells is sensitive and reliable tool for diagnosis and disease monitoring in patients with monoclonal gammopathy. Circulating PCs (cPCs) in peripheral blood (PB) after autologous hematopoietic stem cell transplantation is a marker of high-risk disease in patients with plasma cell myeloma. We assessed the utility of quantification of cPCs using flow cytometry for risk stratification in newly diagnosed plasma cell myeloma patients in the era of novel agents. Methods PB and bone marrow (BM) aspirates of 85 newly diagnosed patients with symptomatic plasma cell myeloma from August 2013 to July 2014 were analyzed by five-color flow cytometry using monoclonal antibodies against CD45, CD19, CD56, CD38, and CD138. The gating strategy employed first used the expression of CD38 and CD138 to identify plasma cells among 100,000 to 200,000 events. cPCs in PB was determined according to the patient's specific immunophenotype of neoplastic PCs in BM. Results The median age of the patient population was 68 years (45~87) and 58% were female. Median follow-up duration was 19.2 months. Six out of 85 patients (7%) did not show cPCs. Among 79 patient (93%) who had detectable cPCs, the median cPCs was 0.09% (0.006~3.612%). Patients without cPCs or cPCs under 0.05% were assigned to low cPCs group (n=32, 38%) and others to high cPCs group (n=53, 62%) according to receiver operating characteristics analysis. High cPCs group showed higher level of BM neoplastic PCs detected by both methodologys of morphology and flow cytometry (P=0.002, 0.033, respectively), higher BM cellularity (P=0.011), higher serum M protein level (P=0.013), lower hemoglobin (P=0.008), and lower platelet level (P=0.034) than low cPCs group. High cPCs group was associated with adverse cytogenetics such as t(4;14) and monosomy 13 (P=0.008), and CD45 negative immunophenotype (P=0.007). In survival analysis, high cPCs presented shorter overall survival (OS) than low cPCs group (P=0.013) (Fig. 1). It was independent with patient age and cytogenetic risks (P =0.011). Conclusion By flow cytometry cPCs was detected in most symptomatic plasma cell myeloma patients. Increased cPCs ≥0.05% among PB leukocytes could be an independent prognostic factor showing adverse effect in overall survival in symptomatic plasma cell myeloma patients. Figure 1. Kaplan-Meier survival curve of patients with plasma cell myeloma who showed 0.05% or more circulating plasma cells in peripheral blood and patients with circulating plasma cells less than 0.05%. Figure 1. Kaplan-Meier survival curve of patients with plasma cell myeloma who showed 0.05% or more circulating plasma cells in peripheral blood and patients with circulating plasma cells less than 0.05%. Disclosures No relevant conflicts of interest to declare.

5 Multiple myeloma flow cytometry panel validated for clinical monitoring of patients

2020 ◽  
Vol 8 (Suppl 3) ◽  
pp. A5-A5
Author(s):  
Bevan Gang ◽  
Vicky Sgouroudis ◽  
Virginia Litwin ◽  
Anita Boyapati
Keyword(s):  
Multiple Myeloma ◽  
Flow Cytometry ◽  
Plasma Cell ◽  
Plasma Cells ◽  
Temperature Stability ◽  
Clinical Samples ◽  
Blood Collection ◽  
Significant Heterogeneity ◽  
Flow Cytometric ◽  
Limit Of Quantitation

BackgroundMultiple myeloma (MM) is an incurable plasma cell malignancy with significant heterogeneity in clinical presentation. Plasma cells are antibody-producing cells of lymphoid origin that are resident in secondary lymphoid organs and in the bone marrow (BM). The detection of circulating malignant plasma cells using flow cytometry has also been described in patients with MM. Enumerating and phenotyping malignant plasma cells in the BM and peripheral blood (PB) may be of value when evaluating the presence of MM antigens targeted by therapies before and during treatment and at relapse. To this end, a flow cytometric panel was developed to enumerate and characterize malignant plasma cells and additional immune subsets.MethodsPB and BM aspirates (BMA) were obtained from healthy donors and MM donors who consented to research testing. MM cell lines were also used to spike into donor samples to detect specific antigens (collected in Cyto-Chex® blood collection tubes). Samples were then transferred to TruCount tubes to enumerate immune populations. Fluorescently labeled antibodies directed against CD38, CD138, CD56, CD45, BCMA were evaluated to assess parameters such as time and temperature stability of the reportable immune populations by monitoring the frequencies of the populations. In addition, the limit of quantitation, intra- and inter-assay precision were determined.ResultsThe MM Counting Panel was optimized to leverage antigen expression and fluorophore combinations. A gating strategy enabled enumeration of MM cells based on antigens that can be further subdivided based on BCMA expression. Further testing showed that the precision in frequencies and absolute counts of key reportable populations was deemed acceptable (%CV of <30%). The precision was within the acceptance criteria of%CV <30% for populations with =100 cells. Stability testing revealed that samples were more stable at ambient temperature relative to 4oC, with stability being maintained for 48 h post-collection, where at least 85% of reportable immune readouts were stable (%change <30% relative to baseline), for BMA and PB from various donors (healthy and MM).The panel was ultimately deployed for use with clinical samples from MM clinical trials. Clinical data generated from the MM Counting Panel allowed the identification of malignant plasma cell populations in BMA of patients from trial assessing a BCMAxCD3 bi-specific antibody (NCT03761108).ConclusionsA flow cytometric assay to enumerate and identify normal and malignant plasma cells in MM patients was successfully developed. The approach used can be applied to develop assays for other indications in which patients are treated with therapies.

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