A single-tube six-colour flow cytometry screening assay for the detection of minimal residual disease in myeloma

Tute, Ruth M. de; Jack, Andrew; Child, J. Anthony; Morgan, Gareth J.; Owen, Roger G.; Rawstron, Andy C.

doi:10.1038/sj.leu.2404815

Cited by 39 publications

(36 citation statements)

References 11 publications

(15 reference statements)

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“…In 20 cases, the monotypic plasma cells had the distinctive, bright CD38 and CD138 coexpression typically seen in true plasma cell neoplasms such as multiple myeloma (Figure 4a and c). 9,10 In contrast to typical true plasma cell neoplasms, however, the plasma cells in lymphoplasmacytic lymphoma were almost always CD19 and CD45 positive. When compared with the cytoplasmic Ig light chain-restricted B cells, which were identified in the same assay by CD45 and side light scatter gating, the plasma cells frequently had a brighter cytoplasmic Ig expression (26 of 32 cases) and a dimmer expression of CD19 (19 of 32; Figure 4b).…”

Section: Resultsmentioning

confidence: 93%

“…In these analyses, plasma cells are identified through CD38 and CD138 coexpression, and normal and abnormal cells are distinguished through the differential expression of CD19 and CD45 in conjunction with the documentation of cytoplasmic Ig light chain restriction. 9,10 In this study, the results of these high power plasma cell flow cytometric analyses were compared with the bone marrow aspirate and biopsy pathology, immunohistochemistry, B-cell flow cytometric immunophenotyping, and with the serological and clinical features in a group of 35 lymphoplasmacytic lymphoma cases. The goals of this study were to fully characterize the pathological features of marrow involvement by lymphoplasmacytic lymphoma, to determine whether there were pathological features that correlated with the serological and/ or clinical findings of Waldenströ m's macroglobulinemia, and to develop a rational approach to using these tools for diagnosing marrow involvement by lymphoplasmacytic lymphoma.…”

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confidence: 99%

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Novel immunophenotypic features of marrow lymphoplasmacytic lymphoma and correlation with Waldenström's macroglobulinemia

et al. 2009

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Lymphoplasmacytic lymphoma involving the bone marrow can be difficult to diagnose, and pathological features that predict the presence of associated Waldenströ m's macroglobulinemia have yet to be identified. To address these issues, marrow histology, immunohistochemistry, and flow cytometry were studied from 35 lymphoplasmacytic lymphoma cases that had comprehensive clinical assessment for Waldenströ m's macroglobulinemia. In all cases, the plasma cells were analyzed by a novel 6-color flow method. Both immunohistochemistry and flow cytometry were useful in identifying the lymphoid and plasmacytic disease components. In 19 cases, immunohistochemistry revealed an earlier unrecognized pattern of plasma cell infiltration in which they were physically separate from the lymphoid infiltrates. B-cell flow cytometry revealed monotypic cells in 96% of the cases. Approximately half of these were CD5 and/or CD23 positive, although none had features of chronic lymphocytic leukemia, and none of the B cells had flow cytometric features suggesting plasmacytic differentiation. In contrast, highly sensitive 6-color plasma cell flow cytometry revealed monotypic cells in 32 of the 35 cases; in 20 cases, the pattern of CD38 and CD138 coexpression detected was identical to that seen in plasma cell malignancies such as multiple myeloma. In 18 of these 20 lymphoplasmacytic lymphoma cases, these plasma cells were CD19 positive, distinguishing them from those of true plasma cell neoplasms, which are CD19 negative. It is interesting that the two lymphoplasmacytic lymphoma cases with CD19-negative plasma cells had an IgG isotype serum paraprotein. Apart from this, no other pathological correlates of the clinical or laboratory features of symptomatic Waldenströ m's macroglobulinemia were identified.

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Section: Resultsmentioning

confidence: 93%

mentioning

confidence: 99%

Novel immunophenotypic features of marrow lymphoplasmacytic lymphoma and correlation with Waldenström's macroglobulinemia

et al. 2009

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“…This information may also be useful to help define potential prognostic markers in PCM and MGUS progression to PCM as shown in (5,(13)(14)(15)(16)(17)(18)(19)(20)(21). The combination of immunophenotype together with clonality results is important in evaluation of MRD (2,(43)(44)(45) and also to identify new targets for myeloma therapy (5,15,(23)(24)(25)(26)(27)(28)(29)(30)(31)(32)(33)(34)(35)(36)(37). Because of the need to define the normal PC immunophenotype, bone marrow aspirate samples obtained from patients without PCM, MGUS or other PC neoplasms were studied here for their expression of normal PC antigens.…”

Section: Discussionmentioning

confidence: 99%

Multiparameter immunophenotyping by flow cytometry in multiple myeloma: The diagnostic utility of defining ranges of normal antigenic expression in comparison to histology

Cannizzo

Bellio

Sohani

et al. 2010

Cytometry Part B Clinical

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Background: Numerous studies have reported on the immunophenotype of plasma cells (PCs) in monoclonal gammopathy of undetermined significance (MGUS) and in plasma cell myeloma (PCM), but very few have examined the immunophenotype of normal PCs. In this study, an objective definition of normal range of expression for each antigen was found on normal control PCs. Using these new ranges of normal expression (new method) is different from using a static 20% of PCs cut-off for all antigens as described in the literature (traditional method). These newly calculated normal ranges for each antigen were applied to our data, and compared to histologic and immunohistochemical findings.Methods: Bone marrow samples from 46 patients with PC neoplasms and 15 normal controls were studied. A minimum of 100 PC were analyzed for each patient and control sample. An 8-color staining method was applied to study the immunophenotype of PCs, using a BD FACSCanto II.Results: By the new ranges of normality calculated in this study it was determined that different antigens have different level of expression on polyclonal PCs. CD19 correlated with histology by both the traditional and new methods, but had superior correlation by the new method.Conclusions: This report is the first 8-color immunophenotypic study of PCM in which a ''range of normal expression'' for each antigen is defined. This is a critical step to help distinguish between a normal and neoplastic PC immunophenotype and discern which antigens are of diagnostic importance. V C 2010

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“…Such approaches can detect neoplastic cells even when they represent as little as 0.01% of leucocytes. 67 An example of clonality assessment is shown in Figure 1.…”

Section: Clonality Assessmentmentioning

confidence: 99%

Report of the European Myeloma Network on multiparametric flow cytometry in multiple myeloma and related disorders

Rawstron¹,

Órfão²,

Beksaç³

et al. 2008

Haematologica

439

514

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The European Myeloma Network (EMN) organized two flow cytometry workshops. The first aimed to identify specific indications for flow cytometry in patients with monoclonal gammopathies, and consensus technical approaches through a questionnaire-based review of current practice in participating laboratories. The second aimed to resolve outstanding technical issues and develop a consensus approach to analysis of plasma cells. The primary clinical applications identified were: differential diagnosis of neoplastic plasma cell disorders from reactive plasmacytosis; identifying risk of progression in patients with MGUS and detecting minimal residual disease. A range of technical recommendations were identified, including: 1) CD38, CD138 and CD45 should all be included in at least one tube for plasma cell identification and enumeration. The primary gate should be based on CD38 vs. CD138 expression; 2) after treatment, clonality assessment is only likely to be informative when combined with immunophenotype to detect abnormal cells. Flow cytometry is suitable for demonstrating a stringent complete remission; 3) for detection of abnormal plasma cells, a minimal panel should include CD19 and CD56. A preferred panel would also include CD20, CD117, CD28 and CD27; 4) discrepancies between the percentage of plasma cells detected by flow cytometry and morphology are primarily related to sample quality and it is, therefore, important to determine that marrow elements are present in follow-up samples, particularly normal plasma cells in MRD negative cases.

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A single-tube six-colour flow cytometry screening assay for the detection of minimal residual disease in myeloma

Cited by 39 publications

References 11 publications

Novel immunophenotypic features of marrow lymphoplasmacytic lymphoma and correlation with Waldenström's macroglobulinemia

Novel immunophenotypic features of marrow lymphoplasmacytic lymphoma and correlation with Waldenström's macroglobulinemia

Multiparameter immunophenotyping by flow cytometry in multiple myeloma: The diagnostic utility of defining ranges of normal antigenic expression in comparison to histology

Report of the European Myeloma Network on multiparametric flow cytometry in multiple myeloma and related disorders

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