Cytometry Part B: Clinical Cytometry最新文献

{"title":"Spurious CD34 expression in mantle cell lymphoma due to non-specific binding to PerCP-Cy5.5: A diagnostic pitfall.","authors":"Abhilash Krishnan, Senthilkumaran Danassegarane, Arunkumar Lakshmanan, Rakhee Kar","doi":"10.1002/cyto.b.70083","DOIUrl":"https://doi.org/10.1002/cyto.b.70083","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2026-10-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149179757","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Overall concordance may overstate peripheral blood MRD performance in AML remission.","authors":"Ercan Yılmaz, Erdem Topal","doi":"10.1002/cyto.b.70079","DOIUrl":"https://doi.org/10.1002/cyto.b.70079","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2026-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149126922","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluation of a multiple myeloma synthetic cell mimic as an assay control for flow cytometry.","authors":"Dhong Hyun Lee, Mike Liu, Kanwal Palla, Subhanip Biswas, Swetha Pratyusha Gunturu, Brian Kerfs, Kristine Valenteros, Sarah Kotanchiyev, Shari Kaiser, Justine Dell'Aringa","doi":"10.1002/cyto.b.70075","DOIUrl":"https://doi.org/10.1002/cyto.b.70075","url":null,"abstract":"<p><p>The reliability of clinical flow cytometry assays hinges on the availability of robust assay controls that accurately reflect representative samples. Widely used conventional assay controls, such as cell lines, often fail to replicate the phenotypical and biophysical properties of representative primary samples resulting in gating variability and limited confidence in assay performance as certain markers may not be expressed. To address these limitations, we developed a custom synthetic multiple myeloma (MM) cell mimic using Slingshot Biosciences TruCytes® technology. Currently, the MM1.S cell line can be used as an assay control in a flow cytometry panel to evaluate key markers, B cell maturation antigen (BCMA) and G protein-coupled receptor, class C, group 5, member D (GPRC5D), on plasma cells (PCs) and B cells in bone marrow aspirate, for MM clinical trials. Although MM.1S shows strong expression of the key markers BCMA and GPRC5D, it lacks CD45 and CD19 which are needed for PC and B cell identification. We aimed to engineer a MM cell mimic to simulate the scatter properties and antigen density for our fresh MM bone marrow aspirate sample testing. Representative MM bone marrow mononuclear cells (BMMCs) were reviewed to find scatter properties and representative median fluorescence intensity (MFI) of CD45, CD19, CD38, CD138, BCMA, GPRC5D, CD56, and PD-L1 on PCs and B cells. We engineered the MM cell mimic with one population representing PCs and another population representing B cells, based on MM BMMC samples. The MM cell mimic overcame the lack of CD45 and CD19 expression and the divergent scatter and MFI profiles observed in traditional MM cell lines like MM1.S. Qualification studies demonstrated intra- and inter-assay precision (%CV ≤20%), robust inter-instrument reproducibility, post-reconstitution stability up to 7 days at 4°C, and closed-vial storage stability of up to 13 months at -20°C. In summary, these results establish the MM mimics as an improved and alternate assay control to MM1.S, facilitating longitudinal assay performance monitoring and ongoing quality assurance in clinical flow cytometry.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2026-10-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149105409","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Subset-specific and tissue-specific expression of NK-cell markers and killer immunoglobulin-like receptors: Implications for NK-cell clonality assessment.","authors":"Valerie Woodings, Orrin Koepke, Jason E Love, Alireza Torabi, Lianqun Qiu, Jonathan R Fromm, Sindhu Cherian, Afshin Shameli","doi":"10.1002/cyto.b.70078","DOIUrl":"https://doi.org/10.1002/cyto.b.70078","url":null,"abstract":"<p><p>Accurate immunophenotypic evaluation of natural killer (NK)-cell neoplasms is challenging because NK-cells lack somatically rearranged antigen receptors, are comprised of phenotypically distinct subsets, and exhibit tissue-specific subset distribution. We developed a flow cytometry (FC) panel for NK-cell immunophenotypic analysis and clonality assessment and applied it to 120 specimens, including normal peripheral blood (PB), bone marrow (BM), lymph node (LN), and body fluids, as well as a series of NK-cell neoplasms and NK-cell clones of uncertain significance (NK-CUS). The expression ranges for NK-cell receptors (NKR) including killer immunoglobulin-like receptors (KIR, CD158) in the PB were consistent with those reported previously by other groups. The distribution of NK-cell subsets differed by the specimen type. While PB and BM NK-cells showed predominance of the CD56^dim subset, LN and fluid samples were enriched for the CD56^bright subset, resulting in higher CD94/NKG2A and lower KIR expression compared to PB. Epstein-Barr-virus (EBV)-positive NK/T-cell neoplasms exhibited a relatively consistent CD56^bright CD16^- CD94/NKG2A⁺ CD57^- KIR^- immunophenotype that resembled normal CD56^bright NK-cells, emphasizing the importance of immunophenotypic aberrancies and increased scatter properties rather than restricted KIR expression in diagnosing such neoplasms. NK-large granular lymphocytic leukemia (NK-LGLL) demonstrated a mature CD56^dim phenotype with restricted KIR expression. Twelve cases fulfilled the proposed criteria for NK-CUS, comprising both putative cytomegalovirus (CMV)-related NK-cell clones and phenotypically distinct (likely) CMV-unrelated clones with more pronounced immunophenotypic abnormalities. Our findings suggest that interpretation of NK-cell clonality should take into account NK-cell subset composition and tissue-specific receptor expression. These results provide practical considerations for NK-cell immunophenotypic analysis and contribute to a better understanding of the role of FC in diagnosing such neoplasms.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2026-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149105350","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Old data, new tricks: Comprehensive computational analysis of 10 years of multi-center EuroFlow acute myeloid leukemia diagnostic immunophenotypic data.","authors":"Sarah Bonte, Rosan Olsman, Sofie Van Gassen, Sergio Matarraz, Neus Villamor, Stefan Nierkens, Paula Fernandez, Elaine Sobral da Costa, Carmen-Mariana Aanei, Alberto Orfao, Jacques J M van Dongen, Yvan Saeys, Mattias Hofmans, Vincent H J van der Velden","doi":"10.1002/cyto.b.70077","DOIUrl":"https://doi.org/10.1002/cyto.b.70077","url":null,"abstract":"<p><p>Acute myeloid leukemia (AML) is characterized by high genotypic and immunophenotypic heterogeneity. We collected an extensive dataset containing 5366 flow cytometry files from 885 AML patients, stained with the EuroFlow acute leukemia orientation tube (ALOT) and AML/MDS panel, acquired in a standardized way at eight centers over a period of 10 years. Unsupervised clustering identified groups of patients based on FlowSOM-derived cell population percentages. In addition, we investigated immunophenotypic patterns in World Health Organization (WHO) patient classes and NPM1^mut subclasses, both at the cell population and the individual marker level. Some WHO classes, for example, AML with t(8;21)(q22;q22)/RUNX1::RUNX1T1 or t(15;17)(q24;q21)/PML::RARA, showed homogeneous immunophenotypes. Characterization of maturation arrest using FlowSOM confirmed maturation arrest at early stages in distinct WHO classes. Finally, a machine learning model was trained to predict WHO genetic classes from immunophenotypic data. The model allowed accurate prediction in 77% of cases, reproducible in an independent validation cohort. In conclusion, we show that EuroFlow standardized protocols allow analysis of multi-centric data measured over an extended period of time. Computational analysis demonstrated inter- and intrapatient immunophenotypic heterogeneity and allowed prediction of genetic abnormalities.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2026-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149073951","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A practical difference from the normal approach to evaluate natural killer (NK) cells and NK cell neoplasms by flow cytometry.","authors":"Aaron J Wilk, Afshin Shameli, Erik Ames, Jonathan R Fromm, Bartosz Grzywacz, Min Shi, Sindhu Cherian, Jean Oak","doi":"10.1002/cyto.b.70076","DOIUrl":"https://doi.org/10.1002/cyto.b.70076","url":null,"abstract":"<p><p>Natural killer (NK) cell neoplasms span indolent to highly aggressive entities, and flow cytometric immunophenotyping is central to their detection and classification. However, distinguishing reactive from neoplastic NK cell expansions can be challenging because NK cells lack somatically recombined antigen receptors, and reactive states may produce restricted phenotypes that mimic clonality. This best-practices manuscript outlines a practical, flow cytometry-based approach to NK cell analysis that integrates (i) robust lineage definition and gating, (ii) subset-aware interpretation of CD16/CD56 patterns, (iii) inference of clonality using killer cell immunoglobulin-like receptor (KIR; CD158) repertoire restriction and other NK cell receptors. We highlight key pitfalls-particularly the limited utility of KIR-based clonality inference in CD56^brightCD16^- NK cells and the potential for adaptive NK cell expansions to appear KIR-restricted-and propose an operational reflex algorithm triggered by NK cell expansion and/or defined immunophenotypic abnormalities in screening tubes. Case-based examples illustrate common scenarios, including NK-large granular lymphocytic leukemia, extranodal NK/T-cell lymphoma with circulating CD56^brightCD16^- cells, reactive adaptive NK cell expansions, and concurrent neoplastic and reactive NK clones. This framework supports standardized, reproducible NK cell assessment and improves specificity of flow-based clonality inference in routine clinical practice.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2026-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149036848","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of morphological and immunophenotypic heterogeneity in adult T-cell leukemia/lymphoma and development of a peripheral blood tumor load-based prognostic index.","authors":"Feng Zhang, Huarong Zhou, Fei Gao, Meng Lin, Yanrong Liu, Nainong Li, Chenqing Zhang","doi":"10.1002/cyto.b.70074","DOIUrl":"https://doi.org/10.1002/cyto.b.70074","url":null,"abstract":"<p><p>We evaluated morphologic and immunophenotypic features of adult T-cell leukemia/lymphoma (ATLL) using morphology and flow cytometry (FCM), compared tumor cell proportions in bone marrow (BM) and peripheral blood (PB), and assessed whether these parameters could improve prognostication beyond existing indices for aggressive ATLL. We retrospectively analyzed PB and BM morphology, FCM, and clinical records of individuals newly diagnosed with ATLL at our institution from 2010 to 2024. Morphologic reevaluation was performed in 47 of 58 enrolled individuals, all with leukemic manifestations. ATLL cells showed heterogeneity. Cytoplasmic vacuoles were present in 61.7% of cases, including a previously undescribed pattern of small peripheral vacuoles (38.3%). Nuclear forms included flower-like (66.0%), cerebriform (10.6%), and small indented lymphocyte-like (36.2%) cells. Cells typically expressed CD4, CD25, CD2, and CD5. However, CD25 was negative in 26.3% of cases when assessed with a weakly conjugated fluorochrome. CD8, CD7, and CD56 were consistently absent, CD3 was variable, whereas TRBC1 showed preliminary variability in the limited subset. Both cytology (with more ATLL cells showing abnormal morphology in PB than in BM) and flow cytometry demonstrated a higher tumor burden in PB than in BM. FCM detected BM infiltration more sensitively than conventional BM smears. In this single-center derivation cohort without validation, the PB tumor load prognostic index (PBTL-PI), incorporating PB abnormal lymphocyte percentage (PB-PAL) and corrected calcium, showed excellent apparent discrimination (area under the curve [AUC] = 0.871). PBTL-PI outperformed the Japan Clinical Oncology Group prognostic index in risk stratification (log-rank p = 0.0002 vs. 0.0147) and prognostic accuracy (AUC: 0.871 vs. 0.738, DeLong's p = 0.024). However, these findings are exploratory and require independent validation. ATLL shows morphologic and immunophenotypic heterogeneity. Cytology and flow cytometry both confirmed higher tumor burden in PB (vs. BM). FCM improves the detection of BM infiltration. PBTL-PI may identify high-risk ATLL and guide risk-adapted management.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2026-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149015825","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Clinical Optimized Multiparameter Immunophenotyping Panels: A new publication series in Cytometry Part B.","authors":"Ahmad Al-Attar, Kamila Czechowska","doi":"10.1002/cyto.b.70072","DOIUrl":"https://doi.org/10.1002/cyto.b.70072","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2026-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148975994","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Circulating tumor plasma cells quantification in newly diagnosed transplant ineligible real world multiple myeloma patients: Comparison of single-platform flow cytometry versus next generation flow.","authors":"Elona Saraci, Alessandra Larocca, Roberto Ria, Silvia Mangiacavalli, Giulia Benevolo, Nicola Giuliani, Renato Zambello, Angelo Belotti, Ilaria Rizzello, Antonietta Pia Falcone, Iolanda Donatella Vincelli, Mariagrazia Michieli, Sonia Ronconi, Patrizia Tosi, Maria Letizia Mosca Siez, Alessandro Allegra, Maide Maria Cavalli, Matilde Scaldaferri, Ilaria Fedele, Marco Burdisso, Tania Villanova, Cristina Velluti, Marcello Capriata, Donato Mannina, Anna Maria Cafro, Elisabetta Antonioli, Sonia Morè, Mario Boccadoro, Benedetto Bruno, Sara Bringhen, Mattia D'Agostino","doi":"10.1002/cyto.b.70068","DOIUrl":"https://doi.org/10.1002/cyto.b.70068","url":null,"abstract":"<p><p>Circulating tumor plasma cells (CTCs) at diagnosis are independent prognostic markers in newly diagnosed multiple myeloma (NDMM). Flow cytometry (FC) enables sensitive and cost-effective CTC quantification, but prognostic cut-offs vary across studies. The methodological approach used for quantification may affect the determination of the CTC percentage. In this study we compare head-to-head two FC techniques [single-platform FC versus Next Generation Flow (NGF)] to measure CTC levels in NDMM patients. Methodological differences between the two techniques can be found in sample processing (NGF includes fixation and permeabilization steps that are not required for CTC quantification by single-platform FC) and analytical sensitivity, which is higher for NGF than for single-platform FC. The percentage of CTCs was simultaneously assessed with both techniques in the peripheral blood (PB) of 34 patients enrolled in the REAL MM trial (NCT03829371) at screening. Single-platform FC detected CTCs in 29/34 patients (85.3%) patients, while NGF detected CTCs in 31/34 patients (91.2%), including 2 cases that were negative by single-platform FC. The CTC percentage measured using the NGF technique (median 0.0049%, range 0%-0.377%) was significantly lower than that obtained using single-platform FC (median 0.009%, range 0%-0.59%) (p < 0.0001). Indeed, quantifying CTCs with FC after fixation and permeabilization (comparably to NGF) led to results that were similar to NGF. In conclusion, NGF demonstrated higher sensitivity than single-platform FC to detect CTCs at diagnosis in MM patients. Employing standardized techniques like NGF may avoid variability across different laboratories and centers.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2026-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148967074","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Stimulus-specific interpretation in DHR testing for chronic granulomatous disease.","authors":"Nezihe Köker Özer, Ercan Yılmaz, Mustafa Yavuz Köker","doi":"10.1002/cyto.b.70073","DOIUrl":"https://doi.org/10.1002/cyto.b.70073","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":" ","pages":""},"PeriodicalIF":2.9,"publicationDate":"2026-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148963186","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}