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Vol. 47. Núm. S3.
HEMO 2025 / III Simpósio Brasileiro de Citometria de Fluxo
(Outubro 2025)
Vol. 47. Núm. S3.
HEMO 2025 / III Simpósio Brasileiro de Citometria de Fluxo
(Outubro 2025)
5
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T-CELL CLONES OF UNCERTAIN SIGNIFICANCE/ T-CUS
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Maria Daniela Holthausen Perico, Renata da Silva Kalfeltz, Andressa de Oliveira Martin Wagner, Anise Osório Ferri, Franciani Turra Costella Delagnello, Liziani Crestani Bortoluzzi, Paula Gomes Back Prim
Hematology and Hemotherapy Center of Santa Catarina (HEMOSC), Cell Markers Laboratory, Florianopolis, SC, Brazil
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Vol. 47. Núm S3

HEMO 2025 / III Simpósio Brasileiro de Citometria de Fluxo

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Female patient, 56 years old, with leukocytosis and lymphocytosis and Lymphoma suspicion - Peripheral blood sample with white blood count 23,200 cells / TCD8+ restriction (TCD4/TCD8 ratio 0.1:1). Panel, Immunophenotyping and Gating Strategy. Triage of all the samples was analyzed in the LST tube. The TCRCBeta 1 tube was designed based on EuroFlow’s Orientation Tube for Immunodeficiencies (PIDOT), using CD3, CD4, CD8 and CD45 as “backbones”, CD27 and CD45RA to discriminate T-cell subsets, CD16 and CD56 to exclude NK cells, and including TCRCBeta1 in the PE channel. Results: 59% T Cells: *52.5% TCD8+: 39.4% TD (terminally differentiated) phenotype with increased absolute levels (9,141 cells/μL - normal age range 8-500) CD7dim CD27(-) CD45RA-/+ TCRCBeta1+5% (monoclonal); 0.7% Naive CD45RA+ CD27+ TCRCBeta1+30% (polyclonal), 1.2% CM/TM (central/terminal memory) CD45RA(-) CD27+ TCRCBeta1+19% (polyclonal); 9.7% EM (effector memory) phenotype with increased absolute levels (2,250 cells/μL - normal age range 2-323) CD45RA(-) CD27+ TCRCBeta1+5% (monoclonal). *4.9% TCD4+ TCRCBeta1+44% (polyclonal). Immunophenotype: CD45++ CD3+ CD8+ CD38dim CD45RA-/+ CD45RO-/+ CD2+ CD5++ CD7+FR TCR ALPHA-BETA+ PERFORIN+ TCRCBETA1 ++5%; Negative expression: CD25 CD26 CD27 CD28 TCL1 CCR7 TCR GAMMA-DELTA CD11C CD30 CD16 CD56 CD94. Morphology: In the analyzed smear, 32% of atypical medium-sized lymphoid cells were observed, with a globose nucleus, generally eccentric, with poorly condensed chromatin with an outline of a nucleolus, and a moderately basophilic, polarized and granular cytoplasm. The inicial diagnostic hypothesis was T-Lymphoma, NOS (the authors judged that there were no sufficient NK antigens expressed to characterize T-LGL). However, further investigation showed the patient had no identifiable T Cell lymphoma, and she eventually was diagnosed with Thrombotic Thrombocytopenic Purpura, with classic symptoms and laboratory confirmation. Since the implementation of TRBC1 for detection of abnormal T-cells, persistent clonal expansion of large granular lymphocytes (T-cell clones of uncertain significance/ T-CUS) have been reported in a variety of clinical conditions, such as hematological neoplasms and autoimmunity. The incidence of T-CUS increases with age, and probably represents reactive T-cell small clones (so called immunoclones), with less than 20% of total lymphocytes or 400 cells/mm3, highly prevalent in patients without T-cell malignancy. Most reactive T-CUS cases are CD8+ or CD4+/CD8+ double-positive.

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T-CUS in pink, normal TCD4+ cells in green and normal TCD8+ cells in dark blue.

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Hematology, Transfusion and Cell Therapy
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