Cytometry Part B: Clinical Cytometry最新文献

{"title":"Automated quantification of measurable residual disease in chronic lymphocytic leukemia using an artificial intelligence-assisted workflow","authors":"Alexandre Bazinet,&nbsp;Alan Wang,&nbsp;Xinmei Li,&nbsp;Fuli Jia,&nbsp;Huan Mo,&nbsp;Wei Wang,&nbsp;Sa A. Wang","doi":"10.1002/cyto.b.22116","DOIUrl":"10.1002/cyto.b.22116","url":null,"abstract":"<p>Detection of measurable residual disease (MRD) in chronic lymphocytic leukemia (CLL) is an important prognostic marker. The most common CLL MRD method in current use is multiparameter flow cytometry, but availability is limited by the need for expert manual analysis. Automated analysis has the potential to expand access to CLL MRD testing. We evaluated the performance of an artificial intelligence (AI)-assisted multiparameter flow cytometry (MFC) workflow for CLL MRD. We randomly selected 113 CLL MRD FCS files and divided them into training and validation sets. The training set (<i>n</i> = 41) was gated by expert manual analysis and used to train the AI model. We then compared the validation set (<i>n</i> = 72) MRD results obtained by the AI-assisted analysis versus those by expert manual analysis using the Pearson correlation coefficient and Bland–Altman plot method. In the validation set, the AI-assisted analysis correctly categorized cases as MRD-negative versus MRD-positive in 96% of cases. When comparing the AI-assisted analysis versus the expert manual analysis, the Pearson <i>r</i> was 0.8650, mean bias was 0.2237 log₁₀ units, and the 95% limit of agreement (LOA) was ±1.0282 log₁₀ units. The AI-assisted analysis performed sub-optimally in atypical immunophenotype CLL and in cases lacking residual normal B cells. When excluding these outlier cases, the mean bias improved to 0.0680 log₁₀ units and the 95% LOA to ±0.2926 log₁₀ units. An automated AI-assisted workflow allows for the quantification of MRD in CLL with typical immunophenotype. Further work is required to improve performance in atypical immunophenotype CLL.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2023-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10758725","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":"Increased IFN-γ+ and TNF-α+ mucosal-associated invariant T cells in patients with aplastic anemia","authors":"Xiaohui Chen,&nbsp;Yuping Zhang,&nbsp;Yikai Zhang,&nbsp;Yue Zhang,&nbsp;Shunqing Wang,&nbsp;Zhi Yu,&nbsp;Xiaoen Liu,&nbsp;Guixuan Huang,&nbsp;Lixing Guo,&nbsp;Xueqin Li,&nbsp;Xianfeng Zha,&nbsp;Yangqiu Li,&nbsp;Bo Li","doi":"10.1002/cyto.b.22115","DOIUrl":"https://doi.org/10.1002/cyto.b.22115","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Aplastic anemia (AA) is known as an autoimmune disease in which T cell activation is aberrant. It has been reported that unconventional T cells, mucosal-associated invariant T (MAIT) cells, play an important role in several autoimmune diseases, but it is unclear if they are involved in AA.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>In this study, we for the first time analyzed the proportions, phenotypes, and cytokine properties of MAIT cells in AA by flow cytometry.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>We found that the percentage of circulating MAIT cells was generally higher for CD3⁺, CD8⁺, and CD8⁻ T cells in AA patients compared with healthy individuals. Moreover, the percentage of IL-18Rα-, NKG2D-, IFN-γ-, and TNF-α- positive MAIT cells was also significantly higher in AA patients. In addition, the percentage of IFN-γ⁺ CD3⁺ or TNF-α⁺CD8⁻ MAIT cells had a significant negative correlation with the absolute neutrophil count.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>We present the first observation of MAIT cells in patients with AA. MAIT cells are associated with a higher frequency of IFN-γ and TNF-α production and may contribute to the pathogenesis of AA.</p>\u0000 </section>\u0000 </div>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22115","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50150342","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Alignment, segmentation and neighborhood analysis in cyclic immunohistochemistry data using CASSATT","authors":"Asa A. Brockman,&nbsp;Rohit Khurana,&nbsp;Todd Bartkowiak,&nbsp;Portia L. Thomas,&nbsp;Shamilene Sivagnanam,&nbsp;Courtney B. Betts,&nbsp;Lisa M. Coussens,&nbsp;Christine M. Lovly,&nbsp;Jonathan M. Irish,&nbsp;Rebecca A. Ihrie","doi":"10.1002/cyto.b.22114","DOIUrl":"10.1002/cyto.b.22114","url":null,"abstract":"<p>Cyclic immunohistochemistry (cycIHC) uses sequential rounds of colorimetric immunostaining and imaging for quantitative mapping of location and number of cells of interest. Additionally, cycIHC benefits from the speed and simplicity of brightfield microscopy, making the collection of entire tissue sections and slides possible at a trivial cost compared to other high dimensional imaging modalities. However, large cycIHC datasets currently require an expert data scientist to concatenate separate open-source tools for each step of image pre-processing, registration, and segmentation, or the use of proprietary software. Here, we present a unified and user-friendly pipeline for processing, aligning, and analyzing cycIHC data - Cyclic Analysis of Single-Cell Subsets and Tissue Territories (CASSATT). CASSATT registers scanned slide images across all rounds of staining, segments individual nuclei, and measures marker expression on each detected cell. Beyond straightforward single cell data analysis outputs, CASSATT explores the spatial relationships between cell populations. By calculating the log odds of interaction frequencies between cell populations within tissues and tissue regions, this pipeline helps users identify populations of cells that interact—or do not interact—at frequencies that are greater than those occurring by chance. It also identifies specific neighborhoods of cells based on the assortment of neighboring cell types that surround each cell in the sample. The presence and location of these neighborhoods can be compared across slides or within distinct regions within a tissue. CASSATT is a fully open source workflow tool developed to process cycIHC data and will allow greater utilization of this powerful staining technique.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22114","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9947383","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Flow cytometric assessment for minimal/measurable residual disease in B lymphoblastic leukemia/lymphoma in the era of immunotherapy","authors":"Xueyan Chen,&nbsp;Qi Gao,&nbsp;Mikhail Roshal,&nbsp;Sindhu Cherian","doi":"10.1002/cyto.b.22113","DOIUrl":"10.1002/cyto.b.22113","url":null,"abstract":"<p>Minimal/measurable residual disease (MRD) is the most important independent prognostic factor for patients with B-lymphoblastic leukemia (B-LL). MRD post therapy has been incorporated into risk stratification and clinical management, resulting in substantially improved outcomes in pediatric and adult patients. Currently, MRD in B-ALL is most commonly assessed by multiparametric flow cytometry and molecular (polymerase chain reaction or high-throughput sequencing based) methods. The detection of MRD by flow cytometry in B-ALL often begins with B cell antigen-based gating strategies. Over the past several years, targeted immunotherapy directed against B cell markers has been introduced in patients with relapsed or refractory B-ALL and has demonstrated encouraging results. However, targeted therapies have significant impact on the immunophenotype of leukemic blasts, in particular, downregulation or loss of targeted antigens on blasts and normal B cell precursors, posing challenges for MRD detection using standard gating strategies. Novel flow cytometric approaches, using alternative strategies for population identification, sometimes including alternative gating reagents, have been developed and implemented to monitor MRD in the setting of post targeted therapy.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-01-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9561739","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":"Noel Warner Ph.D.","authors":"Frederic Preffer,&nbsp;Jack Dunne,&nbsp;Diether Recktenwald,&nbsp;Andrew Blidy,&nbsp;Lewis Lanier,&nbsp;Joe Trotter,&nbsp;Michael J. Daley","doi":"10.1002/cyto.b.22112","DOIUrl":"10.1002/cyto.b.22112","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9222979","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":"Where diagnosis for myelodysplastic neoplasms (MDS) stands today and where it will go: The role of flow cytometry in evaluation of MDS","authors":"Wei Wang,&nbsp;Joseph D. Khoury","doi":"10.1002/cyto.b.22110","DOIUrl":"10.1002/cyto.b.22110","url":null,"abstract":"<p>Myelodysplastic neoplasms (MDS) are clonal hematopoietic neoplasms defined by cytopenias and morphologic dysplasia. This definition distinguishes MDS from clonal hematopoiesis of indeterminate potential (CHIP) and clonal cytopenia of undetermined significance (CCUS) wherein a patient harbors a somatic mutation involving a myeloid malignancy-associated gene in the absence of cytopenia and, in case of CCUS, dysplasia (Khoury et al., <span>2022</span>). As such, the diagnostic framework for MDS relies on the detection of morphologic dysplasia in bone marrow cells coupled with evidence of clonality in patients with cytopenia.</p><p>While the features of dysplasia in hematopoietic lineages have been long established, assessment is subjective and limited by inter-observer variability particularly in cases with mild morphologic changes. In addition, several non-neoplastic conditions give rise to dysplastic changes that can overlap markedly with those seen in MDS. Examples of such conditions include nutritional deficiencies, toxins, and exposure to various treatments. In view of the challenges inherent in dysplasia assessment, demonstration of clonality in a patient with cytopenia is critical to establishing a diagnosis of MDS, especially in lower-risk disease with no excess of blasts. Establishing clonality has long rested on detection of cytogenetic abnormalities using conventional karyotyping and/or fluorescence in situ hybridization (FISH). Beyond establishing evidence of clonality, cytogenetic studies also play an important role in risk stratification (Greenberg et al., <span>2012</span>), and some cytogenetic abnormalities correlate with distinct morphological and clinical features such as those of <i>MDS with low blasts and 5q deletion</i>. Yet, the problem of using cytogenetic abnormalities in the diagnostic workup of MDS is that they are detected in only up to 50% of MDS cases. In cases with normal cytogenetic findings, other tools are needed to demonstrate clonality and establish the diagnosis of MDS. Mutation profiling using next-generation sequencing (NGS) demonstrates somatic mutations in 80%–90% of MDS cases (Ogawa, <span>2019</span>), providing substantial value in diagnostic assessment and establishing the presence of clonality. Here too, beyond establishing evidence of clonality, mutation profiling studies also play an important role in risk stratification, and some gene abnormalities are required to diagnose certain MDS types with defining genetic abnormalities such as <i>MDS with low blasts and SF3B1 mutation</i> and <i>MDS with biallelic TP53 inactivation</i> (Bernard et al., <span>2022</span>; Khoury et al., <span>2022</span>; Malcovati et al., <span>2020</span>). However, a persistent caveat is that CHIP and CCUS can also occur in otherwise healthy elderly individuals (Jaiswal et al., <span>2014</span>), requiring caution in interpreting the significance of certain mutations, especially those that are well known age-related mutat","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2022-12-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22110","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9222974","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"CD19 negative diffuse large B-cell lymphoma presenting in leukemic phase","authors":"Thulasi Raman Ramalingam,&nbsp;Ragavi Uthayasuriyan,&nbsp;Vikram Prabhakar,&nbsp;Lakshman Vaidhyanathan","doi":"10.1002/cyto.b.22109","DOIUrl":"10.1002/cyto.b.22109","url":null,"abstract":"Diffuse large B cell lymphoma (DLBCL) is the most common non-Hodgkins lymphoma affecting elderly individuals. It is an aggressive disease, but the outcomes have quite improved after the addition of monoclonal antibodies to the chemotherapy regimen. DLBCL in leukemic phase (L-DLBCL) is extremely rare compared with other mature B cell neoplasms. L-DLBCL can mimic acute lymphoblastic leukemia and immunophenotyping (IPT)","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2022-12-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10410020","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":"Multiparameter flow cytometry in the evaluation of myelodysplasia: Analytical issues","authors":"Anna Porwit,&nbsp;Marie C. Béné,&nbsp;Carolien Duetz,&nbsp;Sergio Matarraz,&nbsp;Uta Oelschlaegel,&nbsp;Theresia M. Westers,&nbsp;Orianne Wagner-Ballon,&nbsp;Shahram Kordasti,&nbsp;Peter Valent,&nbsp;Frank Preijers,&nbsp;Canan Alhan,&nbsp;Frauke Bellos,&nbsp;Peter Bettelheim,&nbsp;Kate Burbury,&nbsp;Nicolas Chapuis,&nbsp;Eline Cremers,&nbsp;Matteo G. Della Porta,&nbsp;Alan Dunlop,&nbsp;Lisa Eidenschink-Brodersen,&nbsp;Patricia Font,&nbsp;Michaela Fontenay,&nbsp;Willemijn Hobo,&nbsp;Robin Ireland,&nbsp;Ulrika Johansson,&nbsp;Michael R. Loken,&nbsp;Kiyoyuki Ogata,&nbsp;Alberto Orfao,&nbsp;Katherina Psarra,&nbsp;Leonie Saft,&nbsp;Dolores Subira,&nbsp;Jeroen te Marvelde,&nbsp;Denise A. Wells,&nbsp;Vincent H. J. van der Velden,&nbsp;Wolfgang Kern,&nbsp;Arjan A. van de Loosdrecht","doi":"10.1002/cyto.b.22108","DOIUrl":"https://doi.org/10.1002/cyto.b.22108","url":null,"abstract":"<p>Multiparameter flow cytometry (MFC) is one of the essential ancillary methods in bone marrow (BM) investigation of patients with cytopenia and suspected myelodysplastic syndrome (MDS). MFC can also be applied in the follow-up of MDS patients undergoing treatment. This document summarizes recommendations from the International/European Leukemia Net Working Group for Flow Cytometry in Myelodysplastic Syndromes (ELN <i>i</i>MDS Flow) on the analytical issues in MFC for the diagnostic work-up of MDS. Recommendations for the analysis of several BM cell subsets such as myeloid precursors, maturing granulocytic and monocytic components and erythropoiesis are given. A core set of 17 markers identified as independently related to a cytomorphologic diagnosis of myelodysplasia is suggested as mandatory for MFC evaluation of BM in a patient with cytopenia. A myeloid precursor cell (CD34⁺CD19⁻) count &gt;3% should be considered immunophenotypically indicative of myelodysplasia. However, MFC results should always be evaluated as part of an integrated hematopathology work-up. Looking forward, several machine-learning-based analytical tools of interest should be applied in parallel to conventional analytical methods to investigate their usefulness in integrated diagnostics, risk stratification, and potentially even in the evaluation of response to therapy, based on MFC data. In addition, compiling large uniform datasets is desirable, as most of the machine-learning-based methods tend to perform better with larger numbers of investigated samples, especially in such a heterogeneous disease as MDS.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2022-12-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22108","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50147358","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Human leukocyte antigen-B27 typing by flow cytometry: Comparison of three CE-IVD methods","authors":"Louis Waeckel,&nbsp;Anne Kennel,&nbsp;Anne-Emmanuelle Berger,&nbsp;Claude Lambert","doi":"10.1002/cyto.b.22102","DOIUrl":"10.1002/cyto.b.22102","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>The human leukocyte antigen B27 (HLA-B27), associated with chronic inflammatory diseases is thus widely performed for diagnostic purposes. Genotyping by molecular biology is the current gold standard for HLA-B27 typing, but cheaper and faster flow cytometry methods have been developed.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>In this report, we compare analytical performances of three CE-IVD flow cytometry kits: IOTest™ and DuraClone B27™ from Beckman Coulter and BD HLA-B27™ from BD Biosciences on a Navios™ cytometer as compared to molecular biology.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Routine analyses could conclude for HLA-B27 in197 patients (23.2%) and was not conclusive for 66 patients (7%, 8%). The experience revealed the needs to complete IOTest™ with a lineage marker (like CD3-APC) and a standardization procedure in detection of fluorescence. Comparative analysis considering 23/44 non-conclusive samples as negative, pointed out a 100% sensitivity and specificity of IOTest™ in determining genetically proved HLA-B27. Specificity of the BD HLA-B27^TM kit was 94% (two false positives) sticking to the manufacturer instructions but raised to 100% and 94% sensitivity with a cutoff at 8.5 (225 on FACSdiva's scale).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Discussion</h3>\u0000 \u0000 <p>The DuraClone B27™ specificity was poor using the manufacturer cutoff but raised to 100% with a 8.0 cutoff instead of 15.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>The three flow cytometry kits displayed satisfying performances but need adjustments. The DuraClone B27™ kit seems to be the best while the IOTest™ kit is not conclusive in 8% of cases. In most cases the use of molecular biology can be spared, but genotyping remains essential for patients whose HLA-B27 status cannot be determined with confidence by flow cytometry.</p>\u0000 </section>\u0000 </div>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2022-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22102","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40710972","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Optimization of monocyte gating to quantify monocyte subsets for the diagnosis of chronic myelomonocytic leukemia","authors":"Rebeca Jurado,&nbsp;Maria Huguet,&nbsp;Blanca Xicoy,&nbsp;Marta Cabezon,&nbsp;Ari Jimenez-Ponce,&nbsp;David Quintela,&nbsp;Cristina De La Fuente,&nbsp;Minerva Raya,&nbsp;Esther Vinets,&nbsp;Jordi Junca,&nbsp;Joaquim Julià-Torras,&nbsp;Lurdes Zamora,&nbsp;Albert Oriol,&nbsp;Jose-Tomas Navarro,&nbsp;Xavier Calvo,&nbsp;Marc Sorigue","doi":"10.1002/cyto.b.22106","DOIUrl":"10.1002/cyto.b.22106","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>The presence of &gt;94% classical monocytes (MO1, CD14++/CD16-) in peripheral blood (PB) has an excellent performance for the diagnosis of chronic myelomonocytic leukemia (CMML). However, the monocyte gating strategy is not well defined. The objective of the study was to compare monocyte gating strategies and propose an optimal one.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>This is a prospective, single center study assessing monocyte subsets in PB. First, we compared monocyte subsets using 13 monocyte gating strategies in 10 samples. Then we developed our own 10 color tube and tested it on 124 patients (normal white blood cell counts, reactive monocytosis, CMML and a spectrum of other myeloid malignancies). Both conventional and computational (FlowSOM) analyses were used.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Comparing different monocyte gating strategies, small but significant differences in %MO1 and percentually large differences in %MO3 (nonclassical monocytes) were found, suggesting that the monocyte gating strategy can impact monocyte subset quantification. Then, we designed a 10-color tube for this purpose (CD45/CD33/CD14/CD16/CD64/CD86/CD300/CD2/CD66c/CD56) and applied it to 124 patients. This tube allowed proper monocyte gating even in highly abnormal PB. Computational analysis found a higher %MO1 and lower %MO3 compared to conventional analysis. However, differences between conventional and computational analysis in both MO1 and MO3 were globally consistent and only minimal differences were observed when comparing the ranking of patients according to %MO1 or %MO3 obtained with the conventional versus the computational approach.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>The choice of monocyte gating strategy appears relevant for the monocyte subset distribution test. Our 10-color proposal allowed satisfactory monocyte gating even in highly abnormal PB. Computational analysis seems promising to increase reproducibility in monocyte subset quantification.</p>\u0000 </section>\u0000 </div>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2022-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10034941","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}