Cytometry Part B: Clinical Cytometry最新文献

{"title":"Integration of T-cell clonality screening using TRBC-1 in lymphoma suspect samples by flow cytometry","authors":"Felipe Castillo,&nbsp;Constanza Morales,&nbsp;Biserka Spralja,&nbsp;Joaquín Díaz-Schmidt,&nbsp;Mirentxu Iruretagoyena,&nbsp;Daniel Ernst","doi":"10.1002/cyto.b.22147","DOIUrl":"10.1002/cyto.b.22147","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>The diagnosis of T-cell non-Hodgkin lymphomas (NHL) is challenging. The development of a monoclonal antibody specific for T-cell receptor β constant region 1 (TRBC1) provides an alternative to discriminate clonal T cells. The aim of this study was to evaluate the diagnostic potential of an anti-TRBC1 mAb for the identification of T-NHL.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>We performed a cross-sectional diagnostic analytic study of samples tested for lymphoma. All samples sent for lymphoma screening were first evaluated using the standard Euroflow LST, to which a second additional custom-designed T-cell clonality assessment tube was added CD45/TRBC1/CD2/CD7/CD4/TCRγδ/CD3. Flow cytometry reports were compared with morphological and molecular tests.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Fifty-nine patient samples were evaluated. Within the T-cell population, cut-off percentages in the CD4+ cells were from 29.4 to 54.6% and from 23.9 to 52.1% in CD8+ cells. Cut-off ratios in CD4+ T cells were from 0.33 to 1.1, and in CD8+ cells between 0.22 and 1.0. Using predefined normal cut-off values, 18 of 59 (30.5%) samples showed a restricted expression of TRBC1. A final diagnosis of a T-NHL was confirmed clinically and/or by histopathological studies in 15 of the 18 cases (83.3%). There were no cases of T-NHL by morphology/IHC with normal TRBC1 expression. Non-neoplastic patient samples behaved between predefined TRBC1 cut-off values.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>Expression of TRBC1 provides a robust method for T-cell clonality assessment, with very high sensitivity and good correlation with complementary methods. TRBC1 can be integrated into routine lymphoma screening strategies via flow cytometry.</p>\u0000 </section>\u0000 </div>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138444237","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 alternative processing approach to increase CD138 intensity in flow cytometric analysis of plasma cells","authors":"Deepak Kumar,&nbsp;F. N. U. Kiran,&nbsp;Amanda Wheeler,&nbsp;Rory Dellamano,&nbsp;Richard D. Hammer","doi":"10.1002/cyto.b.22149","DOIUrl":"10.1002/cyto.b.22149","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Surface median immunofluorescence intensity (MFI) of plasma cells antigens, particularly CD138, by flow cytometry underestimates plasma cell populations when compared with that estimated by morphological assessment on Wright's-stained slides. CD138 MFI using traditional sample preparation methods for flow cytometric analysis is often dim and difficult to interpret due to multiple factors. This becomes critical when diagnosing and accurately classifying plasma cell dyscrasias.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>In this study, we analyzed 280 flow cytometric results collected from 2016 to 2022 for CD38 and CD138 MFI on bone marrow aspirates performed by two different methods of sample processing—traditional method of lyse-wash and the alternative method of lyse-no-wash.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Visual examination of histograms showed a clear advantage to CD138 expression intensity with the no-wash method. Although no significant difference was observed in CD38 MFI between the two techniques (<i>p</i> = 0.3), considerable improvement was observed in CD138 MFI with the lyse-no-wash technique of sample processing compared with the conventional method (<i>p</i> = 0.003).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>We concluded that the method of lyse-no-wash is superior to traditional methods especially when it comes to handling bone marrow aspirate samples for plasma cell immunophenotyping. This alternate technique increases the sensitivity of flow cytometry to detect plasma cells resulting in bright and crisp signal intensity for surface CD138. This technique may be particularly advantageous when analyzing low tumor burden such as minimal residual disease.</p>\u0000 </section>\u0000 </div>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-11-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138444236","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":"Phenotype and oxidative burst of low-density neutrophil subpopulations are altered in common variable immunodeficiency patients","authors":"Peter Slanina,&nbsp;Julie Stichova,&nbsp;Veronika Bosakova,&nbsp;Iva Staniczkova Zambo,&nbsp;Marcela Hortova Kohoutkova,&nbsp;Petra Laznickova,&nbsp;Zita Chovancova,&nbsp;Jiri Litzman,&nbsp;Terezie Plucarova,&nbsp;Jan Fric,&nbsp;Marcela Vlkova","doi":"10.1002/cyto.b.22150","DOIUrl":"10.1002/cyto.b.22150","url":null,"abstract":"<p>Common variable immunodeficiency disorder (CVID) is the most common form of primary antibody immunodeficiency. Due to low antibody levels, CVID patients receive intravenous or subcutaneous immunoglobulin replacement therapy as treatment. CVID is associated with the chronic activation of granulocytes, including an increased percentage of low-density neutrophils (LDNs). In this study, we examined changes in the percentage of LDNs and the expression of their surface markers in 25 patients with CVID and 27 healthy donors (HD) after in vitro stimulation of whole blood using IVIg. An oxidative burst assay was used to assess the functionality of LDNs. CVID patients had increased both relative and absolute LDN counts with a higher proportion of mLDNs compared to iLDNs, distinguished based on the expression of CD10 and CD16. Immature LDNs in the CVID and HD groups had significantly reduced oxidative burst capacity compared to mature LDNs. Interestingly we observed reduced oxidative burst capacity, reduced expression of CD10 after stimulation of WB, and higher expression of PD-L1 in mature LDNs in CVID patients compared to HD cells. Our data indicate that that the functional characteristics of LDNs are closely linked to their developmental stage. The observed reduction in oxidative burst capacity in mLDNs in CVID patients could contribute to an increased susceptibility to recurrent bacterial infections among CVID patients.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-11-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22150","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138298613","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":"IgE-mediated bleomycin hypersensitivity: Evidence from drug-reactive T lymphocytes","authors":"Didier Ebo,&nbsp;Michiel Beyens,&nbsp;Alessandro Toscano,&nbsp;Christel Mertens,&nbsp;Jessy Elst,&nbsp;Vito Sabato","doi":"10.1002/cyto.b.22146","DOIUrl":"10.1002/cyto.b.22146","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"107590474","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":"BCR::ABL1 fusion gene positive de novo acute myeloid leukemia with coexistence of NRAS mutation and presented with a peculiar CD58 positive immunophenotype","authors":"Xueya Zhang,&nbsp;Xizhe Guo","doi":"10.1002/cyto.b.22152","DOIUrl":"10.1002/cyto.b.22152","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134648632","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 cytometric analysis of CD34+CD38− cells; cell frequency and immunophenotype based on CD45RA expression pattern","authors":"Shumpei Mizuta,&nbsp;Makoto Iwasaki,&nbsp;Noriko Bandai,&nbsp;Saya Yoshida,&nbsp;Asami Watanabe,&nbsp;Hiroshi Takashima,&nbsp;Takeshi Ueshimo,&nbsp;Kazuhiro Bandai,&nbsp;Kensuke Fujiwara,&nbsp;Naoko Hiranuma,&nbsp;Yusuke Koba,&nbsp;Takahito Kawata,&nbsp;Akira Tamekane,&nbsp;Mitsumasa Watanabe","doi":"10.1002/cyto.b.22148","DOIUrl":"10.1002/cyto.b.22148","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Introduction</h3>\u0000 \u0000 <p>The CD34⁺CD38⁻ population in bone marrow includes hematopoietic stem/progenitor cells. Recently, in acute myeloid leukemia, the focus has shifted to flow cytometry analysis targeting CD34⁺CD38⁻ leukemic cells due to their effectiveness in minimal/measurable residual disease detection and prognosis prediction. Nevertheless, the immunophenotype and cell frequency of these cells in the bone marrow, in the absence of leukemic cells, remains unknown. We aimed to evaluate detailed characteristics of CD34⁺CD38⁻ cells in both normal and leukemic cells by flow cytometry.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>We compared the cell frequency and immunophenotype of the CD34⁺CD38⁻ fraction in the following groups: patients with idiopathic thrombocytopenic purpura and malignant lymphoma as controls (<i>n</i> = 17), post-treatment patients without abnormal blasts (<i>n</i> = 35), and patients with myeloid malignancies (<i>n</i> = 86). The comparison was based on the presence or absence of CD45RA expression, a marker commonly used to prospectively isolate lymphoid-primed cell populations within the CD34⁺CD38⁻ fraction.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The CD34⁺CD38⁻CD45RA⁺ cell population exhibited a significant expansion in bone marrow without leukemic cells 1 month after cord blood transplantation and in various type of myeloid malignancies, compared to the control group (<i>p</i> &lt; 0.01). Continuous CD45RA expression and notable expansion of the CD34⁺CD38⁻CD45RA⁻ population were exclusively observed in myelodysplastic syndrome-related diseases. The CD34⁺CD38⁻CD45RA⁺ population displayed frequent expression of various markers in both leukemic and non-leukemic cells, in contrast to the CD34⁺CD38⁻CD45RA⁻ population.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>The CD34⁺CD38⁻ fraction should be carefully evaluated considering the nature of normal hematopoietic precursor cells, their cell frequency and immunophenotype, including CD45RA expression pattern, for improving the accuracy of myeloid malignancy diagnosis.</p>\u0000 </section>\u0000 </div>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71479197","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—September 2023","authors":"Professor Frederic I. Preffer","doi":"10.1002/cyto.b.22145","DOIUrl":"10.1002/cyto.b.22145","url":null,"abstract":"<p>[Color figure can be viewed at wileyonlinelibrary.com]</p><p>Despite the reality that my entire career has been spent devoted to flow cytometry, it is undeniable to me that the gold standard remains an image of the cell, and when visualized within tissue, the architectural relationships between cells. Thus my keen interest in bringing the Amnis technology into my research laboratory years ago and my current enthusiasm for the cell-imaging capacities of more recent technologies such as those from Becton-Dickinson (Cell-View), ThermoFisher (Attune CytPix) and other imaging flow cytometers currently or soon available. In this issue I am therefore extremely interested in Cyclic Analysis of Single-Cell Subsets and Tissue Territories (CASSATT) which identifies scanned slide images through multiple staining rounds, segments nuclei and assesses marker expression on each detected cell (Brockman et al., <span>2023</span>). Cyclic immunochemistry (cycIHC) exploits multiple rounds of immunostaining and imaging for mapping and locating cells of interest with the speed and simplicity of brightfield microscopy, making the collection of entire tissue sections and slides possible. CASSATT employs registered scanned slide images across all rounds of staining, segments individual nuclei, and measures marker expression on each detected cell. It further explores the spatial relationships between cell populations and the odds of interaction frequencies between cell populations within tissue regions, helping to identify cells that interact or do not interact. In this report, a test dataset of six glioblastoma tissue sections were cyclically stained for eight biomarkers for a total of 48 scanned slide images. The authors describe an efficient workflow that provided answers to questions commonly encountered in discovery research in tissue specimens, such as the overall abundance of cells of interest within a tissue section as well as whether groups of cells were in close proximity. CASSATT gathered together and streamlined all steps necessary to produce single cell expression information from cycIHC datasets and did so utilizing an open source environment. In addition, CASSATT systematically analyzed the spatial relationships between cell populations and via unsupervised algorithms, identified clusters of cell niches.</p><p>The persistence of measurable residual disease (MRD) is a strong indicator for adverse outcomes in acute myeloid leukemia (AML) and has been shown to be a valid surrogate marker for disease-free survival and overall survival, irrespective of patients' age, AML subtype, sample type, time of MRD assessment and MRD detection method (Short et al., <span>2020</span>). Other hematopoietic malignancies such as B- and plasma cell lineages also benefit from MRD diagnostic assays (Chen et al., <span>2023</span>; Gao et al., <span>2023</span>; McMillan et al., <span>2023</span>; Zhou et al., <span>2023</span>). In a study by Wang et al. (<span>2023</span>) th","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22145","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41182275","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":"Analytical assay validation for acute myeloid leukemia measurable residual disease assessment by multiparametric flow cytometry","authors":"Jesse M. Tettero,&nbsp;Naveen Dakappagari,&nbsp;Maaike E. Heidinga,&nbsp;Yvonne Oussoren-Brockhoff,&nbsp;Diana Hanekamp,&nbsp;Anil Pahuja,&nbsp;Kerri Burns,&nbsp;Pavinder Kaur,&nbsp;Zeni Alfonso,&nbsp;Vincent H. J. van der Velden,&nbsp;Jeroen G. te Marvelde,&nbsp;Willemijn Hobo,&nbsp;Jennichjen Slomp,&nbsp;Costa Bachas,&nbsp;Angele Kelder,&nbsp;Kevin Nguyen,&nbsp;Jacqueline Cloos","doi":"10.1002/cyto.b.22144","DOIUrl":"10.1002/cyto.b.22144","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Measurable residual disease (MRD) assessed by multiparametric flow cytometry (MFC) has gained importance in clinical decision-making for acute myeloid leukemia (AML) patients. However, complying with the recent In Vitro Diagnostic Regulations (IVDR) in Europe and Food and Drug Administration (FDA) guidance in the United States requires rigorous validation prior to their use in investigational clinical trials and diagnostics. Validating AML MRD-MFC assays poses challenges due to the unique underlying disease biology and paucity of patient specimens. In this study, we describe an experimental framework for validation that meets regulatory expectations.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Our validation efforts focused on evaluating assay accuracy, analytical specificity, analytical and functional sensitivity (limit of blank (LoB), detection (LLoD) and quantitation (LLoQ)), precision, linearity, sample/reagent stability and establishing the assay background frequencies.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Correlation between different MFC methods was highly significant (<i>r</i> = 0.99 for %blasts and <i>r</i> = 0.93 for %LAIPs). The analysis of LAIP specificity accurately discriminated from negative control cells. The assay demonstrated a LoB of 0.03, LLoD of 0.04, and LLoQ of 0.1%. Precision experiments yielded highly reproducible results (Coefficient of Variation &lt;20%). Stability experiments demonstrated reliable measurement of samples up to 96 h from collection. Furthermore, the reference range of LAIP frequencies in non-AML patients was below 0.1%, ranging from 0.0% to 0.04%.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>In this manuscript, we present the validation of an AML MFC-MRD assay using BM/PB patient specimens, adhering to best practices. Our approach is expected to assist other laboratories in expediting their validation activities to fulfill recent health authority guidelines.</p>\u0000 </section>\u0000 </div>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22144","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41113543","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":"Minimal residual disease assessment in B-cell precursor acute lymphoblastic leukemia by semi-automated identification of normal hematopoietic cells: A EuroFlow study","authors":"Martijn W. C. Verbeek,&nbsp;Beatriz Soriano Rodríguez,&nbsp;Lukasz Sedek,&nbsp;Anna Laqua,&nbsp;Chiara Buracchi,&nbsp;Malicorne Buysse,&nbsp;Michaela Reiterová,&nbsp;Elen Oliveira,&nbsp;Daniela Morf,&nbsp;Sjoerd R. Oude Alink,&nbsp;Susana Barrena,&nbsp;Saskia Kohlscheen,&nbsp;Stefan Nierkens,&nbsp;Mattias Hofmans,&nbsp;Paula Fernandez,&nbsp;Elaine Sobral de Costa,&nbsp;Ester Mejstrikova,&nbsp;Tomasz Szczepanski,&nbsp;Lukasz Slota,&nbsp;Monika Brüggemann,&nbsp;Giuseppe Gaipa,&nbsp;Georgiana Grigore,&nbsp;Jacques J. M. van Dongen,&nbsp;Alberto Orfao,&nbsp;Vincent H. J. van der Velden","doi":"10.1002/cyto.b.22143","DOIUrl":"10.1002/cyto.b.22143","url":null,"abstract":"<p>Presence of minimal residual disease (MRD), detected by flow cytometry, is an important prognostic biomarker in the management of B-cell precursor acute lymphoblastic leukemia (BCP-ALL). However, data-analysis remains mainly expert-dependent. In this study, we designed and validated an Automated Gating &amp; Identification (AGI) tool for MRD analysis in BCP-ALL patients using the two tubes of the EuroFlow 8-color MRD panel. The accuracy, repeatability, and reproducibility of the AGI tool was validated in a multicenter study using bone marrow follow-up samples from 174 BCP-ALL patients, stained with the EuroFlow BCP-ALL MRD panel. In these patients, MRD was assessed both by manual analysis and by AGI tool supported analysis. Comparison of MRD levels obtained between both approaches showed a concordance rate of 83%, with comparable concordances between MRD tubes (tube 1, 2 or both), treatment received (chemotherapy versus targeted therapy) and flow cytometers (FACSCanto versus FACSLyric). After review of discordant cases by additional experts, the concordance increased to 97%. Furthermore, the AGI tool showed excellent intra-expert concordance (100%) and good inter-expert concordance (90%). In addition to MRD levels, also percentages of normal cell populations showed excellent concordance between manual and AGI tool analysis. We conclude that the AGI tool may facilitate MRD analysis using the EuroFlow BCP-ALL MRD protocol and will contribute to a more standardized and objective MRD assessment. However, appropriate training is required for the correct analysis of MRD data.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2023-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22143","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41118047","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":"Whole blood no-lyse no-wash micromethod for the quantitative measurement of monocyte HLA-DR","authors":"Jordi Miatello,&nbsp;Valérie Faivre,&nbsp;Clémence Marais,&nbsp;Mégane Raineau,&nbsp;Didier Payen,&nbsp;Pierre Tissieres","doi":"10.1002/cyto.b.22142","DOIUrl":"10.1002/cyto.b.22142","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Monocyte (m)HLA-DR expression appears to be a potent marker of immunosuppression in critically ill patients. The persistence of low mHLA-DR expression is associated with an increased risk of nosocomial infections and mortality. To adapt this measurement to pediatric requirements and provide extensive 24/7 access, we have developed a whole blood no-lyse no-wash micromethod (MM) and compared it with the standardized method (SM).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>mHLA-DR was quantified by flow cytometry using Quantibrite™ Anti-HLA-DR PE/Monocyte PerCP-Cy™5.5 with either the SM performed in a diagnostic hematology laboratory using manufacturer protocol, or a whole blood no-lyse no-wash MM using an Attune flow cytometer located in the pediatric ICU. Median fluorescence intensity was measured in both techniques and converted to antibodies per cell (AB/C) calibrated with BD Quantibrite™ PE beads. Blood and Quantibrite™ reagent volume used with the MM was reduced by 5-fold compared to SM. In addition to Quantibrite™ Anti-Human HLA-DR PE/Monocyte PerCP-Cy™5.5, MM required anti-CD45 and anti-CD19 labeling.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>We determined the expression of mHLA-DR in 34 patients, 20 adults, and 14 children admitted to ICU. Correlation between MM and SM was excellent (Pearson's correlation: <i>y</i> = 0.8192<i>x</i> + 678.7, <i>r</i> = 0.9270, <i>p</i> &lt; 0.0001). The estimated bias was 2467 ± 1.96 × 3307 AB/C; CI 95% [−4016; +8949].</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>The no-lyse no-wash whole blood microvolume method for measuring mHLA-DR expression allows for simplified sample preparation without compromising accuracy of the data. This method may simplify immune monitoring of critically ill patient by the deployment of a point of care method.</p>\u0000 </section>\u0000 </div>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22142","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10227410","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}