Cytometry Part B: Clinical Cytometry最新文献

{"title":"CD34 and CD117 negative pure erythroid leukemia and phenotypic differences with acute megakaryoblastic leukemia","authors":"Alejandra Altube, Daniela Chelin, Mariela Gomez, Cecilia Malusardi, Dolores Sciaccaluga, Cecilia Cabral, Mariangeles Auat","doi":"10.1002/cyto.b.22160","DOIUrl":"10.1002/cyto.b.22160","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2024-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139377332","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—November 2023","authors":"Professor Alberto Orfao","doi":"10.1002/cyto.b.22154","DOIUrl":"https://doi.org/10.1002/cyto.b.22154","url":null,"abstract":"<p>This new issue of Cytometry B (Clinical Cytometry) consists of five main manuscripts which contain original research in the field of clinical cytometry. A manuscript describing a simple (new) method for preservation of urinary cells for subsequent flow cytometric analyses (Freund et al., <span>2023</span>) opens this issue of the journal. It is followed by three papers related to the application of flow cytometry in the field of acute leukemias. In the first two manuscripts distinct assays for measurable residual disease (MRD) monitoring in acute myeloblastic leukemia (AML) (Tettero et al., <span>2023</span>) and B-cell precursor (BCP) acute lymphoblastic leukemia (ALL) (Arunachalam et al., <span>2023</span>) are (technically and clinically) validated, whereas the third one consists of a comparison and validation of four immunophenotypic scoring systems for the diagnosis of early-T precursor (ETP) acute lymphoblastic leukemia (ALL) (Basavaraju et al., <span>2023</span>). The fifth article in this issue of Cytometry B revisits the application of flow cytometry for HLA-B27 typing through the comparison of 3 CE-IVD certified methods (Waeckel et al., <span>2023</span>). Three letters to the editor complete the contents of the November issue of Cytometry B, in which different aspects of three clinically relevant flow cytometric assays for sepsis (Haem-Rahimi et al., <span>2023</span>), drug-induced hypersensitivity (Ebo et al., <span>2023</span>) and diagnostic screening of acute leukemias (Axler et al., <span>2023</span>), are briefly addressed. In this section, I will summarize the contents and highlight the most relevant contributions of the above papers in four separate blocks related to the fields of (i) the flow cytometric analysis of samples with low cell viability, (ii) the flow cytometric diagnosis and monitoring of acute leukemias, (iii) HLA-B27 typing and (iv) the feasibility to measure HLADR expression levels on stabilized blood monocytes and blood circulating drug-specific T cells in the diagnostic work-up of sepsis and drug-hypersensitivity, respectively.</p><p>Flow cytometry assays used in diagnostic laboratories have mostly focused on blood samples and to a less extent also, in bone marrow and other tissue and body fluid specimens. Despite the high frequency of kidney and urinary tract diseases in the general population, and the frequent need for invasive diagnostic procedures (e.g., kidney biopsy), urine has been one of the less explored and used specimens among the distinct types of body fluids evaluated in (clinical) flow cytometry. Of note, urine samples are frequently obtained for conventional biochemistry assays, including analysis of proteinuria, and for the evaluation of the urine sediment by conventional, for example, cytomorphology. In contrast, flow cytometric analysis of urinary cells (e.g., immune cells, podocytes or epithelial cells) is rarely used in routine diagnostics, despite it has proven to provide valuable infor","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22154","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138739912","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":"Editorial on IVD cellular assay validation","authors":"Bruce H. Davis","doi":"10.1002/cyto.b.22156","DOIUrl":"10.1002/cyto.b.22156","url":null,"abstract":"<p>The article in this issue of Cytometry B on “Standardization of Flow Cytometric Detection of Antigen Expression” by the NCI clinical cytometry group formerly headed by Maryalice Stetler-Stevenson and the NIST group headed by Lili Wang is deserving of not only an accompanying editorial, but special attention by all those readers intending to work in clinical cytometry for the coming decade, as it describes an important future component of diagnostic cellular analysis (Tain et al., <span>2023</span>). Specifically, the ability to measure the antigen (or probe target) expression on well-characterized cell populations will be a vital component of not only monitoring of patients with malignancy, as discussed in the article by Tian et al. herein (Tain et al., <span>2023</span>) but also monitoring of a variety of immune responses or therapeutically altered defined cell populations. A few predictions as to what true antigen quantitation will provide: (1) treatment of sickle cell disease will be adjusted through a standardized measurement of the level of hemoglobin F in F cells (Hgb F containing RBCs) in order to retard the sickling process (De Souza et al., <span>2023</span>); (2) patients with severe infection, cytokine storms and certainly sepsis will be monitored for a combination of activation markers (CD64, CD169, HLA-Dr and others) on neutrophils, monocytes and other cell types for informative and actionable changes regarding the immune status (Bourgoin et al., <span>2020</span>; Davis et al., <span>2006</span>; Davis &amp; Bigelow, <span>2005</span>; Ortillon et al., <span>2021</span>; Schiff et al., <span>1997</span>); (3) Rapid assays for the genetic expression of newly induced targets (CAR-T cells, adenovirus insertion of other targeting receptors, etc.).</p><p>The paper also compares two commonly advocated quantitation methods, PE labeled beads to derive average or median antibody binding capacity (ABC) per cell (Davis et al., <span>1998</span>) vs. single point transformation or ratiometric comparison of the targeted cell population to the CD4 expression on helper T cells using an assumed 40,000 CD4 mAb binding sites per cell (Degheidy et al., <span>2016</span>; Wang et al., <span>2016</span>). Other technical variables the paper convincingly observed is that purified 1:1 PE:antibody preparations give better precision than regular off-the-shelf PE-labeled antibody lots, even if the measured F/P ratio of the off the shelf preparation is close to 1.00. Not surprisingly the study provides quantitative evidence that clone selection does matter and different clones with the same reported target antigen specificity can give variable results, up to nearly a two-fold difference in ABC units and this difference was in no way correctable using the reported F/P ratio of the antibody lot. While the use of 1:1 PE:antibody preparation along with spectrally matched beads for ABC quantitation gave acceptable imprecision with a CV between four instruments","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22156","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138686029","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":"Immunophenotypic portrait of leukemia-associated-phenotype markers in B acute lymphoblastic leukemia","authors":"Emilia Boris,&nbsp;Alexandre Theron,&nbsp;Valentin Montagnon,&nbsp;Nicolas Rouquier,&nbsp;Marion Almeras,&nbsp;Jérôme Moreaux,&nbsp;Caroline Bret","doi":"10.1002/cyto.b.22153","DOIUrl":"10.1002/cyto.b.22153","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Background</h3>\u0000 \u0000 <p>Multiparametric flow cytometry (MFC) is an essential diagnostic tool in B acute lymphoblastic leukemia (B ALL) to determine the B-lineage affiliation of the blast population and to define their complete immunophenotypic profile. Most MFC strategies used in routine laboratories include leukemia-associated phenotype (LAP) markers, whose expression profiles can be difficult to interpret. The aim of our study was to reach a better understanding of 7 LAP markers' landscape in B ALL: CD9, CD21, CD66c, CD58, CD81, CD123, and NG2.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Using a 10-color MFC approach, we evaluated the level of expression of 7 LAP markers including CD9, CD21, CD66c, CD58, CD81, CD123, and NG2, at the surface of normal peripheral blood leukocytes (<i>n</i> = 10 healthy donors), of normal precursor B regenerative cells (<i>n</i> = 40 uninvolved bone marrow samples) and of lymphoblasts (<i>n</i> = 100 peripheral blood samples or bone marrow samples from B ALL patients at diagnosis). The expression profile of B lymphoblasts was analyzed according the presence or absence of recurrent cytogenetic aberrations. The prognostic value of the 7 LAP markers was examined using Maxstat R algorithm.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>In order to help the interpretation of the MFC data in routine laboratories, we first determined internal positive and negative populations among normal leukocytes for each of the seven evaluated LAP markers. Second, their profile of expression was evaluated in normal B cell differentiation in comparison with B lymphoblasts to establish a synopsis of their expression in normal hematogones. We then evaluated the frequency of expression of these LAP markers at the surface of B lymphoblasts at diagnosis of B ALL. CD9 was expressed in 60% of the cases, CD21 in only 3% of the cases, CD58 in 96% of the cases, CD66c in 45% of the cases, CD81 in 97% of the cases, CD123 in 72% of the cases, and NG2 in only 2% of the cases. We confirmed the interest of the CD81/CD58 MFI expression ratio as a way to discriminate hematogones from lymphoblasts. We observed a significant lower expression of CD9 and of CD81 at the surface of B lymphoblasts with a t(9;22)(<i>BCR-ABL</i>) in comparison with B lymphoblasts without any recurrent cytogenetic alteration (<i>p</i> = 0.0317 and <i>p</i> = 0.0011, respectively) and with B lymphoblasts harboring other cytogenetic recurrent abnormalities (<i>p</i> = 0.0032 and <i>p</i> &lt; 0.0001, respectively). B lymphoblasts with t(1;19) at diagnosis significantly overexpressed CD81 when compared with B lymphoblasts with other recurrent cytogenetic abnormalitie","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-11-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cyto.b.22153","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138458473","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":"CD5-negative chronic lymphocytic leukemia: Does this entity really exist?","authors":"Daniel Mazza Matos","doi":"10.1002/cyto.b.22151","DOIUrl":"10.1002/cyto.b.22151","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2023-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138451167","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":"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}