Cytometry Part B: Clinical Cytometry最新文献

{"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}

{"title":"The patterns and diagnostic significance of the lack of surface immunoglobulin light chain on mature B cells in clinical samples for lymphoma workup","authors":"Sara L. Huang,&nbsp;Thomas Fennell,&nbsp;Xia Chen,&nbsp;James Z. Huang","doi":"10.1002/cyto.b.22107","DOIUrl":"10.1002/cyto.b.22107","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Surface immunoglobulin (sIg) light chains are not always detected on mature B cells. This may present as a challenge for clonality determination in clinical flow cytometry.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>To explore the mechanism and diagnostic significance of sIg negative mature B cells, we retrospectively studied 14 cases of sIg negative reactive B-cell lymphocytosis and 89 cases of sIg negative mature B-cell lymphomas. The expression patterns of sIg and cytoplasmic immunoglobulin (cIg) light chains were studied by flow cytometry using both monoclonal and polyclonal antibodies.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>These 14 cases of sIg negative reactive B-cell lymphocytosis were proven to be polytypic based on cytoplasmic light chain studies. In 89 cases of sIg negative mature B-cell lymphomas, we described four distinct patterns of abnormal light chain expression including partial or complete loss of sIg or cIg, suggesting different underlying mechanisms.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>This study represents the first reported series of body or cystic fluids where reactive B cells do not have detectable sIg, arguing strongly against making a diagnosis of B-cell lymphoma based on lack of sIg in mature B cells. Since the lack of sIg does not always predict clonal/neoplastic mature B-cell proliferation, further cIg evaluation should be performed when sIg expression is not detected in mature B cells. The lack of both sIg and cIg in mature B cells may serve as a reliable surrogate clonality/neoplastic marker.</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":"9925296","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 CD22+/CD19−/CD24− progenitors and CD22+/CD19+/CD24− mature B cells: Diagnostic pitfalls for minimal residual disease detection in B-lymphoblastic leukemia","authors":"Ting Zhou,&nbsp;Jeremiah Karrs,&nbsp;Truc Ho,&nbsp;Alyssa Doverte,&nbsp;James N. Kochenderfer,&nbsp;Nirali N. Shah,&nbsp;Constance M. Yuan,&nbsp;Hao-Wei Wang","doi":"10.1002/cyto.b.22104","DOIUrl":"10.1002/cyto.b.22104","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Multiparametric flow cytometry (MFC) has become a powerful tool in minimal residual disease (MRD) detection in B-lymphoblastic leukemia/lymphoma (B-ALL). In the setting of targeted immunotherapy, B-ALL MRD detection often relies on alterative gating strategies, such as the utilization of CD22 and CD24. It is important to depict the full diversity of normal cell populations included in the alternative B-cell gating methods to avoid false-positive results. We describe two CD22-positive non-neoplastic cell populations in the peripheral blood (PB), including one progenitor population of uncertain lineage and one mature B-cell population, which are immunophenotypic mimics of B-ALL.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Using MFC, we investigated the prevalence and phenotypic profiles of both CD22-positive populations in 278 blood samples from 52 patients with B-ALL; these were obtained pre- and post-treatment with CD19 and/or CD22 CAR-T therapies. We further assessed whether these two populations in the blood were exclusively associated with B-ALL or recent anticancer therapies, by performing the same analysis on patients diagnosed with other hematological malignancies but in long-term MRD remission.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The progenitor population and mature B-cell population were detected at low levels in PB of 61.5% and 44.2% of B-ALL patients, respectively. Both cell types showed distinctive and highly consistent antigen expression patterns that are reliably distinguishable from B-ALL. Furthermore, their presence is not restricted solely to B-ALL or recent therapy.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>Our findings aid in building a complete immunophenotypic profile of normal cell populations in PB, thereby preventing misdiagnosis of B-ALL MRD and inappropriate management.</p>\u0000 </section>\u0000 </div>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2022-11-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10032883","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":"Multicenter prospective evaluation of diagnostic potential of flow cytometric aberrancies in myelodysplastic syndromes by the ELN iMDS flow working group","authors":"Wolfgang Kern,&nbsp;Theresia M. Westers,&nbsp;Frauke Bellos,&nbsp;Marie Christine Bene,&nbsp;Peter Bettelheim,&nbsp;Lisa Eidenschink Brodersen,&nbsp;Kate Burbury,&nbsp;Sung-Chao Chu,&nbsp;Matthew Cullen,&nbsp;Matteo Della Porta,&nbsp;Alan Stewart Dunlop,&nbsp;Ulrika Johansson,&nbsp;Sergio Matarraz,&nbsp;Uta Oelschlaegel,&nbsp;Kiyoyuki Ogata,&nbsp;Anna Porwit,&nbsp;Frank Preijers,&nbsp;Katherina Psarra,&nbsp;Leonie Saft,&nbsp;Dolores Subirá,&nbsp;Elisabeth Weiß,&nbsp;Vincent H. J. van der Velden,&nbsp;Arjan van de Loosdrecht","doi":"10.1002/cyto.b.22105","DOIUrl":"10.1002/cyto.b.22105","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Myelodysplastic syndromes (MDS) represent a diagnostic challenge. This prospective multicenter study was conducted to evaluate pre-defined flow cytometric markers in the diagnostic work-up of MDS and chronic myelomonocytic leukemia (CMML).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Thousand six hundred and eighty-two patients with suspected MDS/CMML were analyzed by both cytomorphology according to WHO 2016 criteria and flow cytometry according to ELN recommendations. Flow cytometric readout was categorized ‘non-MDS’ (i.e. no signs of MDS/CMML and limited signs of MDS/CMML) and ‘in agreement with MDS’ (i.e., in agreement with MDS/CMML).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Flow cytometric readout categorized 60% of patients in agreement with MDS, 28% showed limited signs of MDS and 12% had no signs of MDS. In 81% of cases flow cytometric readouts and cytomorphologic diagnosis correlated. For high-risk MDS, the level of concordance was 92%.</p>\u0000 \u0000 <p>A total of 17 immunophenotypic aberrancies were found independently related to MDS/CMML in ≥1 of the subgroups of low-risk MDS, high-risk MDS, CMML. A cut-off of ≥3 of these aberrancies resulted in 80% agreement with cytomorphology (20% cases concordantly negative, 60% positive). Moreover, &gt;3% myeloid progenitor cells were significantly associated with MDS (286/293 such cases, 98%).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>Data from this prospective multicenter study led to recognition of 17 immunophenotypic markers allowing to identify cases ‘in agreement with MDS’. Moreover, data emphasizes the clinical utility of immunophenotyping in MDS diagnostics, given the high concordance between cytomorphology and the flow cytometric readout. Results from the current study challenge the application of the cytomorphologically defined cut-off of 5% blasts for flow cytometry and rather suggest a 3% cut-off for the latter.</p>\u0000 </section>\u0000 </div>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2022-11-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9169766","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":"Flow cytometry in the diagnosis of myelodysplastic syndromes","authors":"Wolfgang Kern,&nbsp;Arjan van de Loosdrecht","doi":"10.1002/cyto.b.22103","DOIUrl":"10.1002/cyto.b.22103","url":null,"abstract":"<p>In hematologic neoplasms, the identification of recurrent disease-associated phenotypic features in general is followed after a prolonged period by the identification of disease-associated and even disease-defining genetic changes based on which specific treatment will be developed. As an example, for chronic myelogenous leukemia (CML) this has taken one and a half century from the first publication on morphology (Virchow, <span>1845</span>) to the first report on a tyrosin-kinase inhibitor (Druker et al., <span>2001</span>). For myelodysplastic syndromes (MDS) the interval between publication of morphologic classification system (Bennett et al., <span>1982</span>) and the most recent WHO classification applying genetic criteria for diagnostics (Khoury et al., <span>2022</span>) the interval has been significantly shorter. For various reasons, the role of immunophenotypic features of MDS and their flow cytometric identification had been neglected for a long time.</p><p>Following pivotal publication on the topic by Denise Wells and Mike Loken (Wells et al., <span>2003</span>) flow cytometrists became increasingly interested and gathered together in 2007 at the MDS Foundation Conference in Florence, Italy, which proved to be the founding hour of the “ELN iMDS flow working group.” Each member of the group has focused their research on assessing the diagnostic application of flow cytometry in MDS. Since 2007, the ELN iMDS flow working group has held yearly meetings to discuss progress in research activities as well as to divise consensus guideline publications aiming to apply flow cytometry in a valid manner for in the diagnostic setting of MDS (Porwit, <span>2014</span>; van de Loosdrecht et al., <span>2009</span>; van de Loosdrecht et al., <span>2013</span>; Westers et al., <span>2012</span>; Westers et al., <span>2017</span>). These meetings allowed not only for continuous scientific discussion between experts from worldwide (i.e., although formed under the roof of the European LeukemiaNet members came from North America, Europe, Asia and Australia) but also for friendships reaching beyond any scientific scope.</p><p>This Special Issue of Clinical Cytometry on the application of flow cytometry in the diagnostic setting of MDS comprises a variety of different papers from the ELN iMDS flow working group reaching from updated consensus guidelines over multi center studies to research results generated by the members of the group. It has been our pleasure to coordinate this group for one and a half decades and we hope the readers of the Special Issue enjoy reading the articles and get stimulated to add their research to the topic.</p><p>Wolfgang Kern declares part ownership of MLL Munich Leukemia Laboratory.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2022-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22103","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9169763","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":"Issue highlights—November 2022","authors":"János Kappelmayer MD PhD","doi":"10.1002/cyto.b.22101","DOIUrl":"10.1002/cyto.b.22101","url":null,"abstract":"<p>In patients with interstitial lung diseases (ILD) broncho-alveolar lavage (BAL) fluid analysis is important in supporting the diagnosis. The suggested site for obtaining diagnostic BAL is the middle lobe of the patient's lungs when it is washed with isotonic saline and the aspirated solution is analyzed for cell concentration and leukocyte subsets. The procedure for obtaining BAL fluid is quite difficult to standardize and the washing and harvesting procedure is usually carried out in five fractions. In ILD like sarcoidosis, idiopathic pulmonary fibrosis, or eosinophilic ILD, BAL fluid analysis can aid in the diagnosis while in other diseases like primary alveolar proteinosis a therapeutic BAL is applied as drug treatment in these cases, which often fail.</p><p>In this issue of <i>Clinical Cytometry</i> two manuscripts from the same group deal with flow cytometric BAL analysis. In the manuscript of Eidhof et al. (<span>2021</span>) the authors compare three methods to stabilize BAL cells for flow cytometric analysis. This procedure has a large practical significance, as cells in the low protein content BAL fluid are quite vulnerable. Samples should be kept on ice and transported to the laboratory and stained within 4 h, otherwise they tend to lyse. In their article, the authors present the comparison of native lavage cells versus stabilized cells utilizing Transfix, Streck Cell Preservative and an in-house method that primarily consists of formaldehyde fixation in buffered conditions containing serum proteins. All three stabilization methods influence the side scatter (SSC) but this does not affect gating. They found a 7-day stability with Transfix and a 28-day stability with their in-house method that can considerably expand the list of laboratories where samples can be transported under the above conditions.</p><p>In their other paper, this group by Mulder et al. (<span>2022</span>) demonstrate the results of their external quality flow cytometric studies. Starting from year 2000 with only 12 participating laboratories they are now involving 42 flow labs. The external QC programme required on one hand the actual analysis of the cytometric items that consisted of leukocyte count, leukocyte differentiation, lymphocyte subsets, T-cell subsets related to peripheral blood and CD103 expression on CD4+ cells with microscopic analysis. On the other hand, the participants received relevant clinical information about each case and were able to report their interpretive comments and their diagnosis as free text. The best matches with the diagnosis were obtained for sarcoidosis, eosinophilic ILD and extrinsic alveolitis, that would advance the diagnosis of these patients.</p><p>In the past decade the implementation of immunotherapy in the treatment of numerous hematological disorders has become a daily practice. The utilization of rituximab has been incorporated into several treatment protocols making it impossible to use anti-CD20 for the identificatio","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2022-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22101","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10686641","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}