Cytometry Part B: Clinical Cytometry最新文献

{"title":"Correction to “Clinical application of flow cytometry in patients with unexplained cytopenia and suspected myelodysplastic syndrome: A report of the European LeukemiaNet International MDS-Flow Cytometry Working Group”","authors":"","doi":"10.1002/cyto.b.22141","DOIUrl":"10.1002/cyto.b.22141","url":null,"abstract":"<p>van de Loosdrecht, A. A., Kern, W., Porwit, A., Valent, P., Kordasti, S., Cremers, E., Alhan, C., Duetz, C., Dunlop, A., Hobo, W., Preijers, F., Wagner-Ballon, O., Chapuis, N., Fontenay, M., Bettelheim, P., Eidenschink-Brodersen, L., Font, P., Johansson, U., Loken, M. R., … Ireland, R. (2023). Clinical application of flow cytometry in patients with unexplained cytopenia and suspected myelodysplastic syndrome: A report of the European LeukemiaNet International MDS-Flow Cytometry Working Group. <i>Cytometry Part B: Clinical Cytometry</i>, <i>104</i>(1), 77–86. https://doi.org/10.1002/cyto.b.22044</p><p>In the originally published article, the Austrian Science Fund, Grant/Award Number: F4704 was incorrectly listed as a funder. This has been corrected in the online version of the article. We apologize for this error.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22141","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10137882","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":"Validation of a 12-color flow cytometry assay for acute myeloid leukemia minimal/measurable residual disease detection","authors":"Sa A. Wang,&nbsp;Jeffrey L. Jorgensen,&nbsp;Shimin Hu,&nbsp;Fuli Jia,&nbsp;Shaoying Li,&nbsp;Sanam Loghavi,&nbsp;Chi Young Ok,&nbsp;Beenu Thakral,&nbsp;Jie Xu,&nbsp;L. Jeffrey Medeiros,&nbsp;Wei Wang","doi":"10.1002/cyto.b.22140","DOIUrl":"10.1002/cyto.b.22140","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Acute myeloid leukemia (AML) minimal/measurable residual disease (MRD) by multicolor flow cytometry is a complex laboratory developed test (LDT), challenging for implementation. We share our experience in the validation of a 12-color AML MRD flow cytometry assay to meet stringent regulatory requirements.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>We worked under the guidelines of the CLSI HL62 publication, illustrated the details of the validation process that was tailored to uniqueness of AML MRD, and tested its clinical validity in 61 patients. The “trueness” was determined by correlating with concurrent molecular genetic testing and follow-up bone marrow examinations.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Under assay specificity, we shared the details of panel design, analysis, and criteria for interpretation and reporting. The assay accuracy was assessed by testing known positive and negative samples and correlating with molecular genetic testing and follow-up bone marrow examination. The limit of detection (LOD) and limit of quantification (LOQ) were validated to a level between 0.01% and 0.1%, varied from the leukemia-associated immunophenotypes (LAIP) and the numbers of events obtained for analysis. Assay linearity, precision and carry over studies all met acceptable criteria. In the clinical validity test, the concordance was 93%, specificity 98% and sensitivity 83%. The most challenging aspects of the assay were the discrimination of pre-leukemic cells (persistent clonal hematopoiesis) or underlying myelodysplastic clones from AML MRD with immunophenotypic switch or subclone selection.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>The validation met all criteria and obtained FDA IDE (investigational device exemption) approval. This study provides ample technical and professional details in setting up the AML MRD flow cytometry assay and illustrates through the example of the “fit for purpose” validation process. We also highlight the need for further characterization of abnormal blasts bearing the potential for AML relapse.</p>\u0000 </section>\u0000 </div>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10040504","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":"Issue highlights—July 2023","authors":"Wolfgang Kern","doi":"10.1002/cyto.b.22138","DOIUrl":"10.1002/cyto.b.22138","url":null,"abstract":"<p>Diagnostic techniques within hematology are becoming more sensitive and measurable residual disease (MRD) is gaining importance as a result—for both diagnosticians and patients. MRD measurement is increasingly used as a study endpoint in clinical trials and also in routine diagnostics—and as a strong predictor of treatment outcome. Currently, more sensitive and specific methods of measurement are in development. Besides molecular assays, flow cytometry is one of the most frequently used methods for MRD assessment due to its shorter turnaround time, its cost-effectiveness, and broader applicability.</p><p>In this issue, several researchers have delved into various aspects of flow cytometry techniques being used for MRD detection within hematological malignancies. Gao et al. introduced a single tube flow cytometry assay with high sensitivity for monitoring MRD in B-lymphoblastic leukemia/lymphoma (B-ALL), independent of specific surface antigen expression like CD19 and CD22. As the authors point out, the development of such assays has become relevant due to the emergence of targeted anti-CD19 and anti-CD22 therapies (Gao, Chen, et al., <span>2023</span>; Gao, Liu, et al., <span>2023</span>). Targeted immunotherapy demonstrated encouraging results in recent years but induces significant changes in the phenotype of leukemic blasts concurrently. Therefore, alternative gating strategies have gained importance and potential CD19 substitutes have been proposed (Chen et al., <span>2023</span>; Mikhailova et al., <span>2022</span>).</p><p>The development of alternative gating strategies is also relevant independently of targeted therapies, as shown by the authors of the next article. In a rare case study, Ramalingam et al. reported a patient with CD19-negative diffuse large B-cell lymphoma (DLBCL). Since CD19 is currently the primary gating marker for B cell neoplasms, its absence may lead to erroneous results and potentially affect therapeutic strategies, so the authors (Ramalingam et al., <span>2022</span>). Challenges for flow cytometry approaches have been studied before (Gao, Chen, et al., <span>2023</span>; Gao, Liu, et al., <span>2023</span>; Huang et al., <span>2023</span>; Martig &amp; Fromm, <span>2022</span>).</p><p>In MRD detection, the avoidance of false positives is crucial as shown by Zhou et al. The authors discussed the pitfalls in MRD detection in B-ALL following targeted immunotherapy, describing the presence of two CD22-positive non-neoplastic cell populations. One progenitor population of uncertain lineage and one mature B-cell population were both immunophenotypic mimics of B-ALL. Zhou et al. concluded that an understanding of these normal cell populations is essential to avoid misinterpretation in MRD assessments and CD19-independent gating strategies, including CD22 and CD24, are key (Zhou et al., <span>2022</span>). Optimizing MRD measurement is at the forefront of numerous studies and alternative antigens (besides CD22 and CD24","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-08-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22138","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10028473","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":"Clinical significance of end of induction measurable residual disease monitoring in B-cell acute lymphoblastic leukemia: A single center experience","authors":"Arun Kumar Arunachalam,&nbsp;Sushil Selvarajan,&nbsp;Thenmozhi Mani,&nbsp;Nancy Beryl Janet,&nbsp;Madhavi Maddali,&nbsp;Sharon Anbumalar Lionel,&nbsp;Uday Kulkarni,&nbsp;Anu Korula,&nbsp;Fouzia N. Aboobacker,&nbsp;Aby Abraham,&nbsp;Biju George,&nbsp;Poonkuzhali Balasubramanian,&nbsp;Vikram Mathews","doi":"10.1002/cyto.b.22139","DOIUrl":"10.1002/cyto.b.22139","url":null,"abstract":"<p>The assessment of measurable residual disease (MRD) has emerged as a powerful prognostic tool for both pediatric and adult acute lymphoblastic leukemia (ALL). This retrospective study aimed to evaluate the prognostic relevance of the end of induction MRD in B-cell acute lymphoblastic leukemia (B ALL) patients. The study included 481 patients who underwent treatment for B ALL between August 2012 and March 2019 and had their MRD at the end of induction assessed by flow cytometry. Baseline demographic characteristics were collected from the patient's clinical records. Event free survival (EFS) and relapse free survival (RFS) were calculated using Kaplan–Meier analysis and survival estimates were compared using the log-rank test. End of induction MRD and baseline karyotype were the strongest predictors of EFS and RFS on multivariate analysis. The EFS was inversely related to the MRD value and the outcomes were similar in patients without morphological remission at the end of induction and patients in remission with MRD ≥1.0%. Even within the subgroups of ALL based on age, karyotype, <i>BCR::ABL1</i> translocation and the treatment protocol, end of induction MRD positive patients had poor outcomes compared to patients who were MRD negative. The study outcome would help draft end of induction MRD-based treatment guidelines for the management of B ALL patients.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-08-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10014115","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":"An ultra-rapid screening method for acute leukemias","authors":"Olof Axler,&nbsp;Filippa Bild,&nbsp;Åsa C. M. Johansson","doi":"10.1002/cyto.b.22137","DOIUrl":"10.1002/cyto.b.22137","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-08-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9957921","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":"Use of a hybrid intelligence decision tree to identify mature B-cell neoplasms.","authors":"Inès Vergnolle,&nbsp;Theo Ceccomarini,&nbsp;Alban Canali,&nbsp;Jean-Baptiste Rieu,&nbsp;François Vergez","doi":"10.1002/cyto.b.22136","DOIUrl":"10.1002/cyto.b.22136","url":null,"abstract":"<p><strong>Background: </strong>Mature B-cell neoplasms are challenging to diagnose due to their heterogeneity and overlapping clinical and biological features. In this study, we present a new workflow strategy that leverages a large amount of flow cytometry data and an artificial intelligence approach to classify these neoplasms.</p><p><strong>Methods: </strong>By combining mathematical tools, such as classification algorithms and regression tree (CART) models, with biological expertise, we have developed a decision tree that accurately identifies mature B-cell neoplasms. This includes chronic lymphocytic leukemia (CLL), for which cytometry has been extensively used, as well as other non-CLL subtypes.</p><p><strong>Results: </strong>The decision tree is easy to use and proposes a diagnosis and classification of mature B-cell neoplasms to the users. It can identify the majority of CLL cases using just three markers: CD5, CD43, and CD200.</p><p><strong>Conclusion: </strong>This approach has the potential to improve the accuracy and efficiency of mature B-cell neoplasm diagnosis.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-08-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9935370","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":"CD19-negative B-cell precursors in bone marrow: A potential mimicker for CD19-negative B-lymphoblastic leukemia by flow cytometry in patients with anti-CD19 treatment","authors":"Weijie Li,&nbsp;Ruth Morgan,&nbsp;Roxanne Nieder,&nbsp;Sahibu Sultan M. Habeebu,&nbsp;G. Doug Myers","doi":"10.1002/cyto.b.22135","DOIUrl":"10.1002/cyto.b.22135","url":null,"abstract":"Novel therapies such as monoclonal antibody or chimeric antigen receptor (CAR) T cell therapy (CAR T) have shown promising results in the management of relapsed/refractory B-lymphoblastic leukemia (B-ALL). CD19, a transmembrane glycoprotein almost always expressed in B-ALL, is a commonly used target by these therapies. Blinatumomab, a bispecific monoclonal antibody T cell engager (BiTE), induces antibody-dependent cellular cytotoxicity against the leukemic cells by binding to CD19 on leukemic cells and CD3 on T cells simultaneously. Tisagenlecleucel (Kymriah), a CAR T targeting CD19, uses patients' own modified T cells to attack CD19+ leukemic cells. The superior efficacy of blinatumomab and tisagenlecleucel has been well demonstrated in relapsed/refractory B-ALL compared with high-dose chemotherapy. Multiparametric flow cytometry (FCM) is commonly used to monitor the therapeutic responses to these treatments. The key strategy for FCM to detect residual or relapsed B-ALL is to find an immature B-cell population phenotypically different from normal B-cell precursors (BCPs, also known as hematogones) in the bone marrow (BM). To investigate if these two anti-CD19 therapies could alter the phenotype of normal BCPs, we retrospectively studied the BCPs in the BM specimens from patients treated with blinatumomab and tisagenlecleucel. We also studied the BCPs from the patients after conventional chemotherapy and hematopoietic stem cell transplantation (HSCT) as a comparison.","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-07-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9879768","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":"Acute leukemia of ambiguous lineage, not otherwise specified with FLT3-ITD mutation and a possible origin in the common lymphoid progenitor","authors":"Fernando Martin-Moro,&nbsp;Jose A. Garcia-Vela","doi":"10.1002/cyto.b.22134","DOIUrl":"10.1002/cyto.b.22134","url":null,"abstract":"The current major classification of acute leukemias is based on the assignment of cellular lineage to the blast population, for which an immunophenotypic analysis generally performed by multiparametric flow cytometry (FCM) is mandatory (Porwit & Béné, 2019). Furthermore, the phenotypic characterization of acute leukemias provides information about the pathogenesis of the disease, although it is sometimes difficult to correlate the blast population compartment with the normal stages of hematopoiesis. Acute leukemias of ambiguous lineage (ALAL) — category included in both the 5th edition of the World Health Organization Classification of Haematolymphoid Tumours (Khoury et al., 2022) and the International Consensus Classification of Myeloid Neoplasms and Acute Leukemias (Arber et al., 2022) — comprise a heterogeneous group of rare and aggressive diseases that includes acute undifferentiated leukemias (AUL) and mixedphenotype acute leukemias (MPAL), thus, ALAL are those that either fail to show evidence of either myeloid, B, or T-lymphoid lineages, or show evidence of commitment to more than one lineage, respectively. The so-called subgroup ALAL, not otherwise specified (NOS) is a catch-all category of diseases that express a combination of markers that do not allow their classification as either other ALAL, and in which a definitive classification along a single lineage is difficult. All those infrequent entities are considered as high-risk diseases and show a poor prognosis when treated with either conventional chemotherapy used for acute myeloid leukemia (AML) or acute lymphoblastic leukemia (ALL). More aggressive approaches such as the FLAG-IDA regimen (fludarabine, cytarabine, granulocyte colony-stimulating factor, and idarubicin) have been explored in patients with ALAL, and","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-05-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9546342","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":"Monocyte HLA-DR expression as an enrollment biomarker in sepsis clinical trials: Evaluation of two sampling tubes and definition of respective clinical thresholds","authors":"Muzhda Haem Rahimi,&nbsp;Filippo Conti,&nbsp;Jean-Francois Llitjos,&nbsp;Aurore Fleurie,&nbsp;Valérie Cerro,&nbsp;Fabienne Venet,&nbsp;Anne-Claire Lukaszewicz,&nbsp;Guillaume Monneret","doi":"10.1002/cyto.b.22133","DOIUrl":"10.1002/cyto.b.22133","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-05-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9522535","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":"Resolving 31 colors on a standard 3-laser full spectrum flow cytometer for immune monitoring of human blood samples","authors":"Linda Hammerich,&nbsp;Yaroslava Shevchenko,&nbsp;Jana Knorr,&nbsp;Wiebke Werner,&nbsp;Alix Bruneau,&nbsp;Frank Tacke","doi":"10.1002/cyto.b.22126","DOIUrl":"10.1002/cyto.b.22126","url":null,"abstract":"<p>Immune monitoring of patients on a single-cell level is becoming increasingly important in various diseases. Due to the often very limited availability of human specimens and our increased understanding of the immune systems there is an increasing demand to analyze as many markers as possible simultaneously in one panel. Full spectrum flow cytometry is emerging as a powerful tool for immune monitoring since 5-laser instruments enable characterization of 40 parameters or more in a single sample. Nevertheless, even if only machines with fewer lasers are available, development of novel fluorophore families enables increasing panel sizes. Here, we demonstrate that careful panel design enables the use of 31-color panels on a 3-laser Cytek® Aurora cytometer for analyzing human peripheral blood leukocytes, without the need for custom configuration and using only commercially available fluorochromes. The panel presented here should serve as an example of a 31-fluorochrome combination that can be resolved on a 3-laser full spectrum cytometer and that can be adapted to comprise other (and possibly more) markers of interest depending on the research focus.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22126","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9489135","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}