• Document: Flow Cytometric Immunophenotypic Analysis of 306 Cases of Multiple Myeloma
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Hematopathology / PLASMA CELL IMMUNOPHENOTYPING Flow Cytometric Immunophenotypic Analysis of 306 Cases of Multiple Myeloma Pei Lin, MD,1* Rebecca Owens,1 Guido Tricot, MD, PhD,2 and Carla S. Wilson, MD, PhD1* Key Words: Syndecan-1; CD138; Plasma cell; Myeloma; Immunophenotype; Flow cytometry DOI: 10.1309/74R4TB90BUWH27JX Abstract Recent advances in flow cytometry (FC) have permitted Bone marrow aspirates from 306 patients with more specific and sensitive evaluations of plasma cell (PC) multiple myeloma were analyzed by flow cytometric populations. Neoplastic PCs traditionally have been identified immunophenotyping. The plasma cells (PCs) were by their CD38+++CD45–/dim staining pattern on FC identified by their characteristic light scatter histograms.1,2 With the use of multiparameter FC and the intro- distribution and reactivity patterns to CD138, CD38, duction of the monoclonal antibody CD138 (syndecan-1), it is and CD45. Monoclonality was confirmed by clear that CD38+++CD45–/dim gating alone might fail to immunoglobulin light chain analysis. The identify myeloma composed largely or partly of CD45+ immunophenotypic profile of the PCs was determined PCs.3,4 CD38 is a nonspecific marker that can be detected on with a panel of antibodies. Moderate to bright hematopoietic stem cells and T and B cells. Neoplastic PCs expression of CD56, CD117, CD20, CD45, and CD52 typically express CD38 at a lower intensity than normal PCs was detected in 71.7%, 17.8%, 9.3%, 8.8%, and 5.2% and might be indistinguishable from contaminating T or B of cases, respectively. These antigens were expressed by cells.5,6 Thus, PC immunophenotyping is best determined by a distinct subpopulation of the PCs in 6.3%, 2.2%, multiparameter FC using at least a 3-color assay that includes 3.7%, 2.9%, and 2.6% of additional cases. CD19 was CD138 (syndecan-1) in the analysis. Syndecan-1 is a trans- negative in more than 99% of cases. The combination of membrane heparan sulfate proteoglycan that typically is CD38 and CD138 was superior to CD38 alone for expressed by PCs and not by T or B cells. 7 Therefore, identifying CD45+ myeloma and separating CD20+ syndecan-1 is considered the most specific marker for PCs. myeloma from B-cell lymphoma. PC The commercially available monoclonal antibody B-B4 recog- immunophenotyping might be useful for detecting nizes an epitope of the syndecan-1 molecule and can be used in minimal residual disease in cases with aberrant antigen FC or immunohistochemical analysis.8 CD138 is reported to expression and for selection of therapeutic agents that be detectable in 60% to 100% of myeloma cases and in 70% to have specific membrane targets. 100% of neoplastic cells in each case.7,9,10 Causes for the wide variation in detection sensitivity are unclear and might be related to technical or biologic factors.8,11 Aberrant immunophenotypes are observed in a majority (87%) of myeloma cases at diagnosis including overexpres- sion of CD56 (62%-75%) and aberrant expression of CD117 (28%) and CD20 (10%).3,5,12,13 While expression of CD56 distinguishes malignant from benign PCs, myeloma without CD56 expression might be associated with more aggressive disease and extramedullary dissemination.14-16 The purposes 482 Am J Clin Pathol 2004;121:482-488 © American Society for Clinical Pathology Downloaded 482 from https://aca

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