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Rapid evaluation of T cell clonality in the diagnostic work-up of mature T cell neoplasms: TRBC1-based flow cytometric assay experience.

ABSTRACT: The identification of mature T cell neoplasms by flow cytometry is often challenging, due to overlapping features with reactive T cells and limitations of currently available T cell clonality assays. The description of an antibody specific for one of two mutually exclusive T cell receptor (TCR) β-chain constant regions (TRBC1) provides an opportunity to facilitate the detection of clonal TCRαβ+ T cells based on TRBC-restriction. Here we prospectively analyzed 14 healthy controls and 63 patients with the flow cytometry protocol currently used for suspected T cell neoplasm implemented with immunostaining targeting TRBC1. Specimens were firstly classified in 3 groups based on clinical records data, laboratory findings and immunophenotypic features. T cell clonality was assessed by TCR Vβ repertoire analysis and the new rapid TRBC1 assay. Results showed that TRBC1 unimodal expression was unequivocally associated with samples presenting with immunophenotypic aberrancies. Moreover, we demonstrated that the use of TRBC1 is useful in solving uncertain cases and confirmed the high sensitivity of the method in identifying small T cell clones of uncertain significance (T-CUS). Finally, we found a high degree of concordance (97%) comparing the currently available clonality assessment methods with the proposed new method. In conclusion, our results provided real-life evidence of the utility of TRBC1 introduction in the flow cytometric clonality evaluation for the routine diagnostic work-up of T cell neoplasms.

SUBMITTER: Capone M

PROVIDER: S-EPMC9530610 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature

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Rapid evaluation of T cell clonality in the diagnostic work-up of mature T cell neoplasms: TRBC1-based flow cytometric assay experience.

Capone Manuela M Peruzzi Benedetta B Palterer Boaz B Bencini Sara S Sanna Alessandro A Puccini Benedetta B Nassi Luca L Salvadori Benedetta B Statello Marinella M Carraresi Alessia A Stefanelli Stefania S Orazzini Chiara C Minuti Barbara B Caporale Roberto R Annunziato Francesco F

Translational oncology 20220930

The identification of mature T cell neoplasms by flow cytometry is often challenging, due to overlapping features with reactive T cells and limitations of currently available T cell clonality assays. The description of an antibody specific for one of two mutually exclusive T cell receptor (TCR) β-chain constant regions (TRBC1) provides an opportunity to facilitate the detection of clonal TCRαβ+ T cells based on TRBC-restriction. Here we prospectively analyzed 14 healthy controls and 63 patients ...[more]

PMID: 36183675

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Project description:Background There is no unified standard data about the sensitivity and specificity regarding flow cytometry analysis of Ki67 expression during lymphoma diagnoses. Objective This evaluated the efficacy of multicolor flow cytometry (MFC) in an estimate of the proliferative activity of B-cell non-Hodgkin lymphoma by comparing the expression of Ki67 using MFC and immunohistochemicals (IHC). Method A total of 559 patients with non-Hodgkin B-cell lymphoma were immunophenotyped using sensitive MFC, of which 517 were newly diagnosed and 42 were transformed lymphomas. Test samples include peripheral blood, bone marrow, various body fluids, and tissues. Through MFC multi-marker accurate gating, abnormal mature B lymphocytes with restricted expression of the light chain were screened. Ki67 was added to determine the proliferation index; the positive rate of Ki67 in tumor B cells was evaluated by cell grouping and internal control. For tissue specimens, MFC and IHC analyses were performed simultaneously to assess the Ki67 proliferation index. Results The positive rate of Ki67 by MFC was correlated with the subtype and aggressiveness of B-cell lymphoma. Ki67 could distinguish indolent lymphomas from aggressive subtypes with a cut-off value of 21.25%, and differentiate transformation from indolent lymphoma with a cut-off value of 7.65%. The expression of Ki67 by MFC (regardless of the type of samples)was highly agreement with the Ki67 proliferative index of tissue samples assessed by pathologic immunohistochemistry. MFC showed a fairly constant negative bias in evaluating tissue or bone marrow samples, compared with IHC. Conclusions Ki67 is a valuable flow marker that can distinguish between indolent and aggressive types of lymphoma and assess whether indolent lymphomas are transformed. Using MFC to evaluate the positive rate of Ki67 is important in clinical settings. MFC has unique advantages in judging the aggressiveness of lymphoma in samples of bone marrow, peripheral blood, pleural and ascites, and cerebrospinal fluid. This is particularly important when tissue samples cannot be obtained, making it an important supplement for pathologic examination.

Dataset Information

Rapid evaluation of T cell clonality in the diagnostic work-up of mature T cell neoplasms: TRBC1-based flow cytometric assay experience.

Publications

Rapid evaluation of T cell clonality in the diagnostic work-up of mature T cell neoplasms: TRBC1-based flow cytometric assay experience.

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