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The 4th revised edition of the WHO Classification of Tumors of Hematopoietic and Lymphoid Tissues (WHO\u2010HAEM) previously defined two categories of HGBCL: the so\u2010called double\u2010hit (DHL) and triple\u2010hit (THL) lymphomas, which were related to forms harboring <jats:italic>MYC<\/jats:italic> and <jats:italic>BCL2<\/jats:italic> and\/or <jats:italic>BCL6<\/jats:italic> rearrangements, and HGBCL, NOS (not otherwise specified), corresponding to entities with intermediate characteristics between diffuse large B\u2010cell lymphoma (DLBCL) and Burkitt lymphoma (BL), without rearrangement of the <jats:italic>MYC<\/jats:italic> and <jats:italic>BCL2<\/jats:italic>, and\/or <jats:italic>BCL6<\/jats:italic> genes. In the 5th edition of the WHO\u2010HAEM, DHL with <jats:italic>MYC<\/jats:italic> and <jats:italic>BCL2<\/jats:italic> rearrangements or THL were reassigned as DLBCL\/HGBCL with <jats:italic>MYC<\/jats:italic> and <jats:italic>BCL2<\/jats:italic> rearrangements (DLBCL\/HGBL\u2010<jats:italic>MYC<\/jats:italic>\/<jats:italic>BCL2<\/jats:italic>), 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\u2010cell lymphoma (DLBCL), and even B acute lymphoblastic leukemia (B\u2010ALL). 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\u2010diagnosed cases and comparing their level of expression with normal peripheral blood (PB) B lymphocytes (<jats:italic>n<\/jats:italic>\u2009=\u200910 samples), tumoral cells from patients diagnosed with B\u2010ALL (<jats:italic>n<\/jats:italic>\u2009=\u200930), BL (<jats:italic>n<\/jats:italic>\u2009=\u200913), or DLBCL (<jats:italic>n<\/jats:italic>\u2009=\u200922). 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\u2010ALL. 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.<\/jats:p>","DOI":"10.1002\/cyto.b.22208","type":"journal-article","created":{"date-parts":[[2024,9,19]],"date-time":"2024-09-19T07:15:25Z","timestamp":1726730125000},"page":"448-464","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["<scp>CD38<\/scp>, <scp>CD39<\/scp>, and <scp>BCL2<\/scp> differentiate disseminated forms of high\u2010grade B\u2010cell lymphomas in biological fluids from Burkitt lymphoma and diffuse large B\u2010cell lymphoma"],"prefix":"10.1002","volume":"106","author":[{"given":"Pauline","family":"Marianini","sequence":"first","affiliation":[{"name":"Department of Biological Hematology, St Eloi Hospital Montpellier University Hospital  Montpellier 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