Immunophenotypic heterogeneity of normal plasma cells: comparison with minimal residual plasma cell myeloma

2012 ◽  
Vol 65 (9) ◽  
pp. 823-829 ◽  
Author(s):  
Dingsheng Liu ◽  
Pei Lin ◽  
Ying Hu ◽  
Yi Zhou ◽  
Guilin Tang ◽  
...  
Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 5036-5036
Author(s):  
Tove Isaacson ◽  
Andrzej Jakubowiak ◽  
Lloyd Stoolman ◽  
Usha Kota ◽  
William Finn ◽  
...  

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.


Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 1903-1903
Author(s):  
Linsheng Zhang ◽  
Sally E. Self ◽  
John Lazarchick

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.


2019 ◽  
Vol 5 (4) ◽  
pp. 20190025
Author(s):  
Margaret Mwania ◽  
Naushad Karim ◽  
Sarah Wambui ◽  
Shamshudin Mohammedali ◽  
Allan Njau

Plasma cell myeloma is a bone marrow disorder characterized by neoplastic proliferation of plasma cells within the bone marrow replacing normal cells. We present a case report of a 25-year-old female with bilateral lower and upper limb pains. She had been seen in various health facilities for the past 2 years with progressively worsening disability. Skeletal survey revealed multiple osteolytic lesions in the appendicular skeleton resembling vanishing bone syndrome. Ultrasound-guided biopsy was done with histological diagnosis of plasma cell myeloma. This case is unique because of the young age at presentation, HIV seropositive status and atypical appearance of the lesions.


2017 ◽  
Vol 94 (3) ◽  
pp. 493-499 ◽  
Author(s):  
Mi Hyun Bae ◽  
Chan-Jeoung Park ◽  
Bo Hyun Kim ◽  
Young-Uk Cho ◽  
Seongsoo Jang ◽  
...  

2012 ◽  
Vol 53 (11) ◽  
pp. 2205-2209 ◽  
Author(s):  
Shoko Nakayama-Ichiyama ◽  
Taiji Yokote ◽  
Yuji Hirata ◽  
Kazuki Iwaki ◽  
Toshikazu Akioka ◽  
...  

2015 ◽  
Vol 139 (10) ◽  
pp. 1276-1280 ◽  
Author(s):  
Michael Keeney ◽  
Jaimie G. Halley ◽  
Daniel D. Rhoads ◽  
M. Qasim Ansari ◽  
Steven J. Kussick ◽  
...  

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.


Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 5039-5039
Author(s):  
Martin Schreder ◽  
Wolfgang Huebl ◽  
Gudrun Koch ◽  
Kathrin Strasser-Weippl ◽  
Niklas Zojer ◽  
...  

Abstract Background: Immunoglobulin-like transcript 2 (ILT2/CD85j) belongs to the Ig superfamily and has homology to the killer cell inhibitory receptors (KIRs). It is expressed on natural killer (NK) cells, monocytes, macrophages, dendritic cells and (naive) B lymphocytes. A differential expression of ILT2 was described for monoclonal gammopathy of undetermined significance (MGUS) and multiple myeloma (MM). Gene expression profiling studies found ILT2 to be downregulated 8.26 fold in myeloma as compared to MGUS, being the most differentially expressed gene between these two subsets. However, as RNA from CD138+ cells was used in this analysis, a varying percentage of normal, non-malignant plasma cells will impact on the results, especially in MGUS cases. Aims: We aimed to delineate ILT2 expression in different plasma cell subsets (normal compared to monoclonal cells) in MGUS and MM and the eventual prognostic impact of a differential expression level. Methods: ILT2 expression was measured by flow cytometry using a PE-conjugated antibody (clone HP-F1, Beckman Coulter) in a series of 30 MGUS patients and 91 myeloma patients. Phenotypically normal and malignant plasma cells were defined by differential expression of CD38, CD45, CD19 and CD56. Expression levels are given as mean fluorescence intensity (MFI) after correction for background staining. Results: ILT2 was not differentially expressed in monoclonal plasma cells from patients with MGUS (MFI median 112.0, range 13.5–274.4) and myeloma (MFI median 96.6, range 0.4–454.5). In contrast, monoclonal cells from MGUS and MM showed a significantly lower expression of ILT2 as compared to phenotypically normal plasma cells in the majority of samples (p=0.007). Results were confirmed by quantitative real time PCR studies in 25 MM patients showing a linear correlation of ILT2 mRNA levels with the intensity of ILT2 protein expression. ILT2 levels did not vary with state of disease and showed no correlation with clinical parameters or prognosis in our series of myeloma patients. Conclusions: In the majority of patients with monoclonal plasma cell disorders, ILT2 seems to be downregulated at an early stage of disease, i.e. upon transformation from a normal plasma cell to the MGUS/MM stage. The expression level of ILT2 in monoclonal plasma cells is neither correlated with the state of disease (MGUS versus newly diagnosed myeloma versus advanced disease) nor to prognosis of myeloma patients or other clinical parameters.


Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 1688-1688
Author(s):  
Soraya Wuilleme ◽  
Nelly Robillard ◽  
Steven Richebourg ◽  
Marion Eveillard ◽  
Laurence Lodé ◽  
...  

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.


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