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Novel multi-parameter flow cytometry sensitively detects phenotypically distinct plasma cell subsets in plasma cell proliferative disorders

Leukemia volume 21, pages 2043–2046 (2007)Cite this article

The detection of abnormal plasma cells (PCs) is a fundamental element in diagnosing plasma cell proliferative disorders (PCPDs) such as multiple myeloma. Flow cytometry (FC) is routinely employed for this purpose; by this method, neoplastic PCs are identified either by restricted patterns of cytoplasmic immunoglobulin (cy-Ig) light chain expression or by aberrant patterns of surface antigen expression. For the latter CD19, CD45 and CD56 are commonly studied based on the observation that abnormal PCs are typically CD56 positive and CD19 and CD45 negative, whereas normal PCs are CD19 and CD45 positive and CD56 negative.1 Although neoplastic PCs can be identified by either of these FC approaches, both have notable shortcomings. It can be difficult to confidently identify abnormal PCs by cy-Ig light chain restriction alone particularly when the number of cells is low, when they are obscured by a background population of normal polytypic PCs, or when the disorders are bi-clonal with subsets positive for κ- and λ-Ig light chain, respectively. While the evaluation of surface antigens avoids many of these pitfalls, the surface immunophenotype is not sufficiently distinctive to allow for the malignant PCs to be identified in up to 10% of PCPDs.2 Furthermore, in all FC analyses it is difficult to formulate a gating strategy which allows for the PCs to be captured, even when real-time (or ‘live’) gating is used. This problem can be exacerbated when evaluating cy-Ig light chain expression as membrane permeabilization required alters the physical properties of the PCs.

Given these limitations of conventional FC, PC evaluation in the clinical laboratory is often supplemented by slide-based immunohistochemical or immunofluorescence (IF) testing. In our laboratory, slide-based IF is performed in conjunction with FC when assessing peripheral blood (PB) and bone marrow (BM) aspirate specimens for potential involvement by a PCPD.3 In this assay, mononuclear cells are isolated by Ficoll gradient. Four-color FC is then performed using two assay tubes each containing antibodies to CD38 and CD45, which are combined with antibodies to cy-Igκ and CD20 in one tube and antibodies to cy-Igλ and CD56 in the other. Cy-Ig light chain-restricted PCs are then identified through a CD38 bright, CD45 dim-to-negative gate. Slide-based IF is performed when FC or serum protein studies indicate the presence of cy-Ig light chain-restricted PCs. This second step is used to confirm the presence of monotypic PCs and also to calculate the PC proliferation fraction by measurement of nuclear incorporation of the thymidine analog bromodeoxyuridine.

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  1. Divisions of Hematopathology and Hematology, Mayo Clinic, Rochester, MN, USA

    W G Morice, C A Hanson, S Kumar, L A Frederick, C E Lesnick & P R Greipp

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  1. W G Morice
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  2. C A Hanson
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  3. S Kumar
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  4. L A Frederick
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  5. C E Lesnick
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Correspondence to W G Morice.

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Morice, W., Hanson, C., Kumar, S. et al. Novel multi-parameter flow cytometry sensitively detects phenotypically distinct plasma cell subsets in plasma cell proliferative disorders. Leukemia 21, 2043–2046 (2007). https://doi.org/10.1038/sj.leu.2404712

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