REVIEW article
Front. Immunol.
Sec. Cancer Immunity and Immunotherapy
Volume 16 - 2025 | doi: 10.3389/fimmu.2025.1642609
Dissecting Immunophenotypic Diversity in Multiple Myeloma via Mass Cytometry: A Call for Harmonized Gating Strategies
Provisionally accepted
- 1University of California Irvine School of Medicine, Irvine, United States
- 2City of Hope National Medical Center, Duarte, United States
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Plasma cell disorders present challenges in phenotypic determination, as they range from monoclonality of plasma cells to multiple myeloma and plasma cell leukemia. According to World Health Organization guidelines, no single aberrant marker is recognized to be uniquely linked to multiple myeloma. The absence of a preset marker panel proven to account for multiple myeloma diversity causes difficulties in diagnosis and clinical research; therefore, the need to create a well-defined panel is urgently needed. For this manuscript, we reviewed the literature on the phenotypic and immunological features that lead to incomplete information and problems in immunophenotyping. We offer proposed solutions for identifying the suitable markers and technology to fill this gap, by using a well-defined gating strategy in a high-dimensional mass cytometry (CyTOF) panel and by next-generation flow cytometry. We analyze pitfalls, starting with sample preparation, selection of the marker panel, gating strategy, cleaning up events, quality control, troubleshooting and validation, and finally, analysis of data. We advance a comprehensive protocol that allows for a detailed analysis of the immunophenotype of myeloma cells. By identifying aberrant markers in the panel, we may be able to facilitate diagnosis and prognosis, ultimately influencing the choice of therapeutic regimens and patients' overall survival.
Keywords: mass cytometry, Flow Cytometry, Multiple Myeloma, Immunophenotyping, Plasma cell, Immune markers, Gating, cyTOF
Received: 06 Jun 2025; Accepted: 29 Jul 2025.
Copyright: © 2025 Singer, Moloudizargari, Sanchez and Pichiorri. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Flavia Pichiorri, City of Hope National Medical Center, Duarte, United States
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