Artifactual Kappa Light Chain Restriction of Marrow Hematogones: A Potential Diagnostic Pitfall in Minimal Residual Disease Assessment of Plasma Cell Myeloma Patients on Daratumumab

2019 ◽  
Vol 98 (1) ◽  
pp. 68-74 ◽  
Author(s):  
Xiu yan Jiang ◽  
Joanne Luider ◽  
Afshin Shameli
Keyword(s):  
Minimal Residual Disease ◽  
Plasma Cell ◽  
Light Chain ◽  
Residual Disease ◽  
Disease Assessment ◽  
Kappa Light Chain ◽  
Plasma Cell Myeloma ◽  
Light Chain Restriction ◽  
Diagnostic Pitfall ◽  
Minimal Residual

scholarly journals Marked Variability in Reported Minimal Residual Disease Lower Level of Detection of 4 Hematolymphoid Neoplasms: A Survey of Participants in the College of American Pathologists Flow Cytometry Proficiency Testing Program

2015 ◽  
Vol 139 (10) ◽  
pp. 1276-1280 ◽  
Author(s):  
Michael Keeney ◽  
Jaimie G. Halley ◽  
Daniel D. Rhoads ◽  
M. Qasim Ansari ◽  
Steven J. Kussick ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Chronic Lymphocytic Leukemia ◽  
Minimal Residual Disease ◽  
Plasma Cell ◽  
Myeloid Leukemia ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Lymphocytic Leukemia ◽  
Plasma Cell Myeloma ◽  
Minimal Residual

Context Flow cytometry is often applied to minimal residual disease (MRD) testing in hematolymphoid neoplasia. Because flow-based MRD tests are developed in the laboratory, testing methodologies and lower levels of detection (LODs) are laboratory dependent. Objectives To broadly survey flow cytometry laboratories about MRD testing in laboratories, if performed, including indications and reported LODs. Design Voluntary supplemental questions were sent to the 549 laboratories participating in the College of American Pathologists (CAP) FL3-A Survey (Flow Cytometry—Immunophenotypic Characterization of Leukemia/Lymphoma) in the spring of 2014. Results A total of 500 laboratories (91%) responded to the supplemental questions as part of the FL3-A Survey by April 2014; of those 500 laboratories, 167 (33%) currently perform MRD for lymphoblastic leukemia, 118 (24%) for myeloid leukemia, 99 (20%) for chronic lymphocytic leukemia, and 91 (18%) for plasma cell myeloma. Other indications include non-Hodgkin lymphoma, hairy cell leukemia, neuroblastoma, and myelodysplastic syndrome. Most responding laboratories that perform MRD for lymphoblastic leukemia reported an LOD of 0.01%. For myeloid leukemia, chronic lymphocytic leukemia, and plasma cell myeloma, most laboratories indicated an LOD of 0.1%. Less than 3% (15 of 500) of laboratories reported LODs of 0.001% for one or more MRD assays performed. Conclusions There is major heterogeneity in the reported LODs of MRD testing performed by laboratories subscribing to the CAP FL3-A Survey. To address that heterogeneity, changes to the Flow Cytometry Checklist for the CAP Laboratory Accreditation Program are suggested that will include new requirements that each laboratory (1) document how an MRD assay's LOD is measured, and (2) include the LOD or lower limit of enumeration for flow-based MRD assays in the final diagnostic report.

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.

scholarly journals Participation in the College of American Pathologists Laboratory Accreditation Program Decreases Variability in B-Lymphoblastic Leukemia and Plasma Cell Myeloma Flow Cytometric Minimal Residual Disease Testing: A Follow-up Survey

2020 ◽  
Author(s):  
Meghan M. Hupp ◽  
Christine Bashleben ◽  
Jolene L. Cardinali ◽  
David M. Dorfman ◽  
William Karlon ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Minimal Residual Disease ◽  
Proficiency Testing ◽  
Plasma Cell ◽  
Residual Disease ◽  
Lymphoblastic Leukemia ◽  
Laboratory Accreditation ◽  
Plasma Cell Myeloma ◽  
Accreditation Program ◽  
Minimal Residual

Context.— Minimal residual disease (MRD) testing by flow cytometry is ubiquitous in hematolymphoid neoplasm monitoring, especially B-lymphoblastic leukemia (B-ALL), for which it provides predictive information and guides management. Major heterogeneity was identified in 2014. Subsequently, new Flow Cytometry Checklist items required documentation of the sensitivity determination method and required lower level of detection (LLOD) inclusion in final reports. This study assesses Laboratory Accreditation Program (LAP) participation and new Checklist items' impact on flow cytometry MRD testing. Objectives.— To survey flow cytometry laboratories about MRD testing for B-ALL and plasma cell myeloma. In particular, enumerate the laboratories performing MRD testing, the proportion performing assays with very low LLODs, and implementation of new Checklist items. Design.— Supplemental questions were distributed in the 2017-A mailing to 548 flow cytometry laboratories subscribed to the College of American Pathologists FL3 Proficiency Testing Survey (Flow Cytometry–Immunophenotypic Characterization of Leukemia/Lymphoma). Results.— The percentage of laboratories performing MRD studies has significantly decreased since 2014. Wide ranges of LLOD and collection event numbers were reported for B-ALL and plasma cell myeloma. Most laboratories determine LLOD by using dilutional studies and include it in final reports; a higher proportion of LAP participants used these practices than nonparticipants. Conclusions.— Several MRD testing aspects vary among laboratories receiving FL3 Proficiency Testing materials. After the survey in 2014, new Checklist items were implemented. As compared to 2014, fewer laboratories are performing MRD studies. While LLOD remains heterogeneous, a high proportion of LAP subscribers follow the new Checklist requirements and, overall, target LLOD recommendations from disease-specific working groups are met.

scholarly journals Standardized assay for assessment of minimal residual disease in blood, bone marrow and apheresis from patients with plasma cell myeloma

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Agnieszka Blum ◽  
Katy Haussmann ◽  
Mathias Streitz ◽  
Stephan Schlickeiser ◽  
Carola Tietze-Buerger ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Minimal Residual Disease ◽  
Plasma Cell ◽  
Residual Disease ◽  
Plasma Cell Myeloma ◽  
Minimal Residual

Daratumumab Interference in Flow Cytometry Producing a False Kappa Light Chain Restriction in Plasma Cells

2020 ◽  
Author(s):  
William Kleinot ◽  
Nadine Aguilera ◽  
Elizabeth L Courville
Keyword(s):  
Monoclonal Antibody ◽  
Flow Cytometry ◽  
Light Chain ◽  
Plasma Cells ◽  
Kappa Light Chain ◽  
Plasma Cell Myeloma ◽  
Light Chain Restriction ◽  
Diagnostic Pitfall ◽  
Flow Cytometric ◽  
B Lineage

Abstract False kappa light chain restriction on hematogones (normal B-lineage precursors) has been described in patients on the therapeutic anti-CD38 monoclonal antibody daratumumab. In this article, we present a novel case report of pseudo-kappa light chain restriction on lambda-restricted neoplastic plasma cells in a patient with progressive plasma cell myeloma while on daratumumab. Flow cytometric technologists and pathologists need to be aware of this potential diagnostic pitfall.

Detection of Minimal Residual Disease of Plasma Cell Myeloma by Five Color Flow Cytometric Immunophenotype Analysis

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 1903-1903
Author(s):  
Linsheng Zhang ◽  
Sally E. Self ◽  
John Lazarchick
Keyword(s):  
Plasma Cell ◽  
Light Chain ◽  
Plasma Cells ◽  
Residual Disease ◽  
Plasma Cell Myeloma ◽  
Immunoglobulin Light Chain ◽  
Color Flow ◽  
Flow Cytometric ◽  
Chain Analysis ◽  
Minimal Residual

Abstract Abstract 1903 Flow cytometry is widely used to identify a monoclonal proliferation of plasma cells through cytoplasmic immunoglobulin light chain analysis, with a better sensitivity than immunocytochemical staining. Recent studies have demonstrated that neoplastic plasma cells express aberrant surface antigens and immunophenotyping of plasma cells by multiparameter flow cytometry is able to reveal neoplastic plasma cells by their surface antigen profile to a level that is meaningful for the detection of prognostically relevant minimal residual disease. In this study, we compare the sensitivity of detecting abnormal plasma cells by five color flow cytometric immunophenotyping and concurrent cytoplasmic immunoglobulin light chain analysis. Multiparameter analysis was performed with cell surface markers CD45, CD38, CD138, CD19, CD20, CD56, CD117, CD27 and CD28. Plasma cells were identified by bright CD38 and CD138 expression. The bone marrow plasma cells from 8 newly diagnosed or recurrent plasma cell myelomas with 10% or more morphologically identifiable plasma cells were initially analyzed. At least one antigen was found to be abnormally expressed in all 8 cases. CD45, CD19, CD56 and CD117 were most useful in recognizing abnormal plasma cells. Both CD45 and CD19 were negative in all 8 cases. CD56 and CD117 were each positive in 6 cases; at least one of them was positive in all 8 cases; and 4 cases co-expressed both antigens. Thirteen additional cases of plasma cell myeloma in clinical remission with less than 10% plasma cells by bone marrow morphology were studied with antibodies to CD38, CD138, CD45, CD56 and CD117 in a single tube. Eleven cases revealed an abnormal immunophenotype, however, immunoglobulin light chain restriction was detected only in 6 cases. Two cases demonstrated normal phenotype and did not show immunoglobulin light chain restriction. Immunoglobulin light chain restriction was not demonstrated in any cases with less than 0.5% bright CD38 plasma cells. In one case with 9% plasma cells by morphologic examination, the immunoglobulin light chain analysis failed to reveal monoclonal proliferation whereas abnormal expression of both CD56 and CD117 was identified in 50% of the bright CD38 and CD138 positive plasma cells, although flow cytometry only detected a total of 0.5% plasma cells. Abnormal phenotype was detected at a level as low as 0.05% plasma cells by flowcytometry, in cases that less than 1% plasma cells were identified by morphologic examination. Our result suggests that 5 color flow cytometric immunophenotyping is a sensitive and practical way to detect minimal residual disease of plasma cell myeloma in patients under clinical remission. Because rare neoplastic plasma cells may not have abnormal surface antigen profile, and the abnormal phenotype may change after chemotherapy, combination with cytoplasmic immunoglobulin light chain analysis may be necessary to increase the sensitivity and specificity. Disclosures: No relevant conflicts of interest to declare.

Consensus guidelines on plasma cell myeloma minimal residual disease analysis and reporting

2015 ◽  
Vol 90 (1) ◽  
pp. 31-39 ◽  
Author(s):  
Maria Arroz ◽  
Neil Came ◽  
Pei Lin ◽  
Weina Chen ◽  
Constance Yuan ◽  
...  
Keyword(s):  
Minimal Residual Disease ◽  
Plasma Cell ◽  
Residual Disease ◽  
Plasma Cell Myeloma ◽  
Consensus Guidelines ◽  
Disease Analysis ◽  
Minimal Residual
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