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  • Title: TCR-Vbeta flow cytometric analysis of peripheral blood for assessing clonality and disease burden in patients with T cell large granular lymphocyte leukaemia.
    Author: Feng B, Jorgensen JL, Hu Y, Medeiros LJ, Wang SA.
    Journal: J Clin Pathol; 2010 Feb; 63(2):141-6. PubMed ID: 20154036.
    Abstract:
    AIMS: T cell large granular lymphocytes (T-LGLs) are commonly increased in reactive conditions as well as T-LGL leukaemia. This differential diagnosis often requires a combined assessment of clonality and tumour burden. In this study we assessed the utility of flow cytometric (FC) analysis of T cell receptor beta chain variable region (TCR-Vbeta) expression by using 24 antibodies reactive to 70% of the TCR-Vbeta repertoire. METHODS: Analyses were performed on peripheral blood samples obtained from 20 patients with a confirmed diagnosis of T-LGL leukaemia and 18 patients without known T cell lymphoproliferative diseases. RESULTS: The results were compared with TCR gene rearrangement status assessed by PCR. By FC analysis, 19/20 T-LGL leukaemia cases were CD3+CD8+ and one case was CD3+CD4+. All the cases demonstrated at least one immunophenotypic aberration, with altered CD5 expression being most frequent. Abnormal Vbeta expression was detected by FC in 19 of 20 (95%) T-LGL leukaemia cases, but in none of the controls; this showed 100% concordance with TCR gene rearrangement studies. In addition to establishing clonality, FC Vbeta analysis enables calculation of absolute numbers of clonal T cells; this is important in monitoring tumour burden after treatment. CONCLUSIONS: It is concluded that FC Vbeta analysis is a fast, reliable and quantitative method that can simultaneously assess T-LGL leukaemia clonality and tumour burden.
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