Accurate detection of the tumor clone in peripheral T-cell lymphoma biopsies by flow cytometric analysis of TCR-Vβ repertoire

Abstract

Multiparametric flow cytometry has proven to be a powerful method for detection and immunophenotypic characterization of clonal subsets, particularly in lymphoproliferative disorders of the B-cell lineage. Although in theory promising, this approach has not been comparably fulfilled in mature T-cell malignancies. Specifically, the T-cell receptor-Vb repertoire analysis in blood can provide strong evidence of clonality, particularly when a single expanded V family is detected. The purpose of this study was to determine the relevance of this approach when applied to biopsies, at the site of tumor involvement. To this end, 30 peripheral T-cell lymphoma and 94 control biopsies were prospectively studied. Vb expansions were commonly detected within CD4 þ or CD8 þ T cells (97% of peripheral T-cell lymphoma and 54% of non-peripheral T-cell lymphoma cases); thus, not differentiating malignant from reactive processes. Interestingly, we demonstrated that using a standardized evaluation, the detection of a high Vb expansion was closely associated with diagnosis of peripheral T-cell lymphoma, with remarkable specificity (98%) and sensitivity (90%). This approach also identified eight cases of peripheral T-cell lymphoma that were not detectable by other forms of immunophenotyping. Moreover, focusing Vb expression analysis to T-cell subsets with aberrant immunophenotypes, we demonstrated that the T-cell clone might be heterogeneous with regard to surface CD7 or CD10 expression (4/11 cases), providing indication on 'phenotypic plasticity'. Finally, among the wide variety of Vb families, the occurrence of a Vb17 expansion in five cases was striking. To our knowledge, this is the first report demonstrating the power of T-cell receptor-Vb repertoire analysis by flow cytometry in biopsies as a basis for peripheral T-cell lymphoma diagnosis and precise T-cell clone identification and characterization.

Flow cytometric pattern recognition of lymph node biopsies with lymphomas that lack lineage characteristics

“…Normal T cells show more phenotypically different subpopulations than normal B cells, not only dependent on their functional status but also on the investigated organ . This variation is also apparent in the Vbeta usage as shown by IPT.…”

“…This variation is also apparent in the Vbeta usage as shown by IPT. Nevertheless two reactive cases showed Vbeta restriction in a series of 94 cases, which is considered as suggestive of T‐NHL . One case was EBV infection related, and the second case was a lymph node with Hodgkin lymphoma.…”

TCR Repertoire Characterization for T Cells Expanded in Response to hRSV Infection in Mice Immunized with a Recombinant BCG Vaccine

“…An explanation that has been proposed is that some TCRs may help to control pathogens since the virus may have less probability to escape via antigenic variation [25]. In noninfectious diseases, such as monoclonal human lymphomas and lymphoid leukemias, TCRVβ rearrangements are used as highly sensitive markers for diagnosis and follow-up of the disease [31,32]. Further, non-Hodgkin's lymphomas patients have been shown to express the same TCRVβ subfamilies as compared to healthy controls [31].…”

Abnormal immunophenotype of the T‐cell‐receptor beta Chain in follicular‐helper T cells of angioimmunoblastic T‐cell lymphoma

“…Vbeta analysis is characteristically performed on subsets of alpha-beta T lymphocytes in blood (1,2), but this study and recent publications have expanded the utility to bone marrow and lymph node biopsies (3,4).…”

“…This Vbeta analysis is characteristically performed on peripheral blood (1,2), but this study and recent publications expand the utility to lymph nodes and bone marrow biopsies of mature T-cell lymphomas (3,4).…”