Cytometry Part B: Clinical Cytometry最新文献

{"title":"CD38, CD39, and BCL2 differentiate disseminated forms of high-grade B-cell lymphomas in biological fluids from Burkitt lymphoma and diffuse large B-cell lymphoma.","authors":"Pauline Marianini,Vanessa Lacheretz-Szablewski,Marion Almeras,Jérôme Moreaux,Caroline Bret","doi":"10.1002/cyto.b.22208","DOIUrl":"https://doi.org/10.1002/cyto.b.22208","url":null,"abstract":"High-grade B-cell lymphomas (HGBCL) represent a heterogeneous group of very rare mature B-cell lymphomas. The 4th revised edition of the WHO Classification of Tumors of Hematopoietic and Lymphoid Tissues (WHO-HAEM) previously defined two categories of HGBCL: the so-called double-hit (DHL) and triple-hit (THL) lymphomas, which were related to forms harboring MYC and BCL2 and/or BCL6 rearrangements, and HGBCL, NOS (not otherwise specified), corresponding to entities with intermediate characteristics between diffuse large B-cell lymphoma (DLBCL) and Burkitt lymphoma (BL), without rearrangement of the MYC and BCL2, and/or BCL6 genes. In the 5th edition of the WHO-HAEM, DHL with MYC and BCL2 rearrangements or THL were reassigned as DLBCL/HGBCL with MYC and BCL2 rearrangements (DLBCL/HGBL-MYC/BCL2), whereas the category HGBCL, NOS remains unchanged. Characterized by an aggressive clinical presentation and a poor prognosis, HGBCL is often diagnosed at an advanced, widespread stage, leading to potential disseminated forms with a leukemic presentation, or spreading to the bone marrow (BM) or other biological fluids. Flow cytometric immunophenotypic study of these disseminated cells can provide a rapid method to identify HGBCL. However, due to the scarcity of cases, only limited data about the immunophenotypic features of HGBCL by multiparametric flow cytometry are available. In addition, identification of HGBCL cells by this technique may be challenging due to clinical, pathological, and biological features that can overlap with other distinct lymphoid malignancies, including Burkitt lymphoma (BL), diffuse large B-cell lymphoma (DLBCL), and even B acute lymphoblastic leukemia (B-ALL). In this study, we aimed to characterize the detailed immunophenotypic portrait of HGBCL, evaluating by multiparametric flow cytometry (MFC) the expression of 26 markers on biological samples obtained from a cohort of 10 newly-diagnosed cases and comparing their level of expression with normal peripheral blood (PB) B lymphocytes (n = 10 samples), tumoral cells from patients diagnosed with B-ALL (n = 30), BL (n = 13), or DLBCL (n = 22). We then proposed a new and simple approach to rapidly distinguish disseminated forms of HGBCL, BL, and DLBCL, using the combination of MFC data for CD38, BCL2, and CD39, the three most discriminative markers explored in this study. We finally confirmed the utility of the scoring system previously proposed by Khanlari to distinguish HGBCL cells from B lymphoblasts of B-ALL. In conclusion, we described a distinct immunophenotypic portrait of HGBCL cells and proposed a strategy to differentiate these cells from other aggressive B lymphoma entities in biological samples.","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142247525","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":"Converting an HLA‐B27 flow assay from the BD FACSCanto to the BD FACSLyric","authors":"Eugene V. Ravkov, Miguel F. Ventura, Swapna Gudipaty, David Ng, Julio C. Delgado, Leo Lin","doi":"10.1002/cyto.b.22206","DOIUrl":"https://doi.org/10.1002/cyto.b.22206","url":null,"abstract":"HLA‐B27 is a major histocompatibility complex (MHC) class I antigen which exhibits strong association (90%) with ankylosing spondylitis. HLA‐B27 detection in patients by flow cytometry is a widely used clinical test, performed on many different flow cytometer models. We sought to develop and validate a test conversion protocol for the HLA‐B27 test performed on the BD FACSCanto to BD's newer FACSLyric flow cytometers. The development and validation experiments were performed using anti‐HLA‐B27*FITC/CD3*PE antibody‐stained whole blood patient specimens. The anti‐HLA‐B27*FITC logarithmic median fluorescence (LMF) results on the BD FACSCanto were converted to median fluorescence intensity (MFI) values on the BD FACSLyric. Clustering of the HLA‐B27 positive and negative values, using a 3rd order polynomial equation, resulted in a conversion of the BD FACSCanto cutoff values, negative (<150 LMF) and positive (≥160 LMF), to negative (<4530 MFI) and positive (≥6950 MFI) on the BD FACSLyric. Accuracy was assessed by comparing the flow results obtained on the BD FACSCanto and BD FACSLyric to a molecular PCR based assay. Additional validation parameters (compensation verification, intra‐ and inter‐assay precision, and instrument comparison) were performed per the recommendations outlined in the Clinical and Laboratory Standards Institute (CLSI) H62 guidelines for validation of flow cytometry assays.","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2024-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142247526","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":"A comprehensive 26‐color immunophenotyping panel to study the role of the gut‐liver axis in chronic liver diseases","authors":"Alix Bruneau, Yaroslava Shevchenko, Frank Tacke, Linda Hammerich","doi":"10.1002/cyto.b.22203","DOIUrl":"https://doi.org/10.1002/cyto.b.22203","url":null,"abstract":"The gut‐liver axis includes the bidirectional communication between the gut and the liver, and thus covers signals from liver‐to‐gut and from gut‐to‐liver. Disruptions of the gut‐liver axis have been associated with the progression of chronic liver diseases, including alcohol‐related and metabolic dysfunction‐associated steatotic liver disease and cholangiopathies. Immune cells and their expression of pattern recognition receptors, activation markers or immune checkpoints might play an active role in the communication between gut and liver. Here, we present a 26‐color full spectrum flow cytometry panel for human cells to decipher the role of circulating immune cells in gut‐liver communication during the progression of chronic liver diseases in a non‐invasive manner, which has been optimized to be used on patient‐derived whole blood samples, the most abundantly available clinical material. Our panel focuses on changes in pattern recognition receptors, including toll‐like receptors (TLRs) or Dectin‐1, and also includes other immunomodulatory molecules such as bile acid receptors and checkpoint molecules. Moreover, this panel can be utilized to follow the progression of chronic liver diseases and could be used as a tool to evaluate the efficiency of therapeutic targets directed against microbial mediators or modulating immune cell activation.","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2024-09-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142209866","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":"CD133 in T-lymphoblastic leukemia is preferentially expressed in early T-phenotype (ETP) and near ETP subtypes.","authors":"Shuyu E, Karen Amelia Nahmod, Beenu Thakral, Wei Wang, Jeffrey L Jorgensen, Sa A Wang","doi":"10.1002/cyto.b.22205","DOIUrl":"https://doi.org/10.1002/cyto.b.22205","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-09-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142139474","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":"Appropriate interpretation of TRBC1-dim positive subsets in T-cell immunophenotyping by flow cytometry.","authors":"Min Shi, Matthew J Weybright, Gregory E Otteson, Dragan Jevremovic, Horatiu Olteanu, Pedro Horna","doi":"10.1002/cyto.b.22204","DOIUrl":"https://doi.org/10.1002/cyto.b.22204","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-09-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142105209","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 role of the primitive marker CD133 in CD34-negative acute myeloid leukemia for the detection of leukemia stem cells.","authors":"Tom Reuvekamp, Luca L G Janssen, Lok Lam Ngai, Jannemieke Carbaat-Ham, Daphne den Hartog, Willemijn J Scholten, Angèle Kelder, Diana Hanekamp, Eliza Wensink, Noortje van Gils, Patrycja Gradowska, Bob Löwenberg, Gert J Ossenkoppele, Arjan A van de Loosdrecht, Theresia M Westers, Linda Smit, Costa Bachas, Jacqueline Cloos","doi":"10.1002/cyto.b.22201","DOIUrl":"https://doi.org/10.1002/cyto.b.22201","url":null,"abstract":"<p><p>The most important reason for dismal outcomes in acute myeloid leukemia (AML) is the development of relapse. Leukemia stem cells (LSCs) are hypothesized to initiate relapse, and high CD34+CD38- LSC load is associated with poor prognosis. In 10% of AML patients, CD34 is not or is low expressed on the leukemic cells (<1%), and CD34+CD38- LSCs are absent. These patients are classified as CD34-negative. We aimed to determine whether the primitive marker CD133 can detect LSCs in CD34-negative AML. We retrospectively quantified 148 CD34-negative patients for proportions of CD34-CD133+ and CD133+CD38- cell fractions in the diagnostic samples of CD34-negative patients in the HOVON102 and HOVON132 trials. No prognostic difference was found between patients with high or low proportions of CD34-CD133+, which is found to be aberrantly expressed in AML. A high level of CD133+CD38- cells was not associated with poor overall survival, and expression in AML was similar to normal bone marrow. To conclude, CD133 is useful as an additional primitive marker for the detection of leukemic blast cells in CD34-negative AML. However, CD133+CD38 alone is not suitable for the detection of LSCs at diagnosis.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142035492","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 assay modifications: Recommendations for method validation based on CLSI H62 guidelines.","authors":"Sara A Monaghan, Steven Eck, Silvia Bunting, Xiangyang X Dong, Robert J Durso, Christele Gonneau, Amanda Hays, Andrea Illingworth, Stacy C League, Eleni Linskens, Megan McCausland, Thomas W McCloskey, Nina Rolf, Min Shi, Paul K Wallace, Virginia Litwin, Wolfgang Kern, George Deeb, Veronica Nash, Horatiu Olteanu","doi":"10.1002/cyto.b.22202","DOIUrl":"https://doi.org/10.1002/cyto.b.22202","url":null,"abstract":"<p><p>The Clinical and Laboratory Standards Institute (CLSI) H62-Validation of Assays Performed by Flow Cytometry guideline, released in 2021, provides recommendations for platform workflow and quality system essentials, instrument setup and standardization, assay development and optimization and fit-for-purpose analytical method validation. In addition, CLSI H62 includes some recommendations for the validation strategies after a validated flow cytometric method has been modified. This manuscript builds on those recommendations and discusses the impact of different types of assay modifications on assay performance. Recommendations regarding which validation parameters to evaluate depending on the type of modification are provided. The impact of assay modification on the assay's intended use is discussed. When recommending minor deviations from the CLSI H62 process for a laboratory-initiated assay revision (e.g., specimen numbers for sensitivity, specificity, or precision studies), a rationale based on expert opinion is provided with the understanding that not every laboratory, assay type, and circumstance can be comprehensively addressed in this paper. These recommendations are meant as a practical recommendation and are not intended to be restrictive, prescriptive, or understood as necessarily sufficient to meet every specific requirement from regulatory bodies (e.g., FDA or New York State Department of Health).</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142008459","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":"Concomitant cutaneous T-cell lymphoma and biclonal B-cell lymphoproliferative disorder.","authors":"Clorinda Derosa, Anna Mestice, Tommasina Perrone, Giuseppe Ingravallo, Michele Troia, Valentina Tabanelli, Raffaele Filotico, Pellegrino Musto","doi":"10.1002/cyto.b.22200","DOIUrl":"https://doi.org/10.1002/cyto.b.22200","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141975333","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":"Mass cytometric single cell immune profiles of peripheral blood from acute myeloid leukemia patients in complete remission with measurable residual disease.","authors":"Øystein Sefland, Stein-Erik Gullaksen, Maria Omsland, Håkon Reikvam, Eivind Galteland, Hoa Thi Tuyet Tran, Signe Spetalen, Satwinder Kaur Singh, Hester J T Van Zeeburg, Arjan A Van De Loosdrecht, Bjørn Tore Gjertsen","doi":"10.1002/cyto.b.22197","DOIUrl":"https://doi.org/10.1002/cyto.b.22197","url":null,"abstract":"<p><p>Measurable residual disease (MRD) is detected in approximately a quarter of AML chemotherapy responders, serving as a predictor for relapse and shorter survival. Immunological control of residual disease is suggested to prevent relapse, but the mechanisms involved are not fully understood. We present a peripheral blood single cell immune profiling by mass cytometry using a 42-antibody panel with particular emphasis on markers of cellular immune response. Six healthy donors were compared with four AML patients with MRD (MRD⁺) in first complete remission (CR1_MRD+). Three of four patients demonstrated a favorable genetic risk profile, while the fourth patient had an unfavorable risk profile (complex karyotype, TP53-mutation) and a high level of MRD. Unsupervised clustering using self-organizing maps and dimensional reduction analysis was performed for visualization and analysis of immune cell subsets. CD57⁺ natural killer (NK)-cell subsets were found to be less abundant in patients than in healthy donors. Both T and NK cells demonstrated elevated expression of activity and maturation markers (CD44, granzyme B, and phosho-STAT5 Y694) in patients. Although mass cytometry remains an expensive method with limited scalability, our data suggest the utility for employing a 42-plex profiling for cellular immune surveillance in whole blood, and possibly as a biomarker platform in future clinical trials. The findings encourage further investigations of single cell immune profiling in CR1_MRD+ AML-patients.</p>","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141792183","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 2024","authors":"Wolfgang Kern,&nbsp;Paul Wallace,&nbsp;Ryan Brinkman","doi":"10.1002/cyto.b.22199","DOIUrl":"10.1002/cyto.b.22199","url":null,"abstract":"","PeriodicalId":10883,"journal":{"name":"Cytometry Part B: Clinical Cytometry","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-07-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141864015","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}