The use of anti-T-cell receptor-Vβ antibodies for the estimation of treatment success and phenotypic characterization of clonal T-cell populations in cutaneous T-cell lymphomas

Sezary syndrome and Mycosis fungoides are the most common forms of cutaneous T-cell lymphomas. To assess the response to different therapies especially in Sezary syndrome, it is helpful to monitor the percentage of circulating tumour cells in the blood. The use of T-cell receptor (TCR)-Vbeta specific monoclonal antibodies provides a suitable tool for detecting Sezary cells. In this study, we analysed the levels of clonal CD4(+) Vbeta(+) cells of seven patients with various treatment modalities using flow cytometry and investigated the immunophenotype of the clonal cells by double staining with a panel of antibodies recognizing lymphatic surface markers. Additionally, a polymerase chain reaction-denaturing gradient gel electrophoresis assay was performed on clonal CD4(+) Vbeta2(+) cells, showing that these cells carry a Vgamma10/11, JgammaP1/2 TCR rearrangement. Follow-up studies revealed close association of the Vbeta(+) clone developmentwith the clinical response to different therapiesinsixpatients. Intwo cases, the CD4(+) Vbeta(+) cells decreased accompanied by partial regression or even complete remission. In four cases, a stable or increasing clonal CD4(+) Vbeta(+) population reflected well a stable or progressing course of the disease. Double staining of Vbeta(+) cells revealed the following pattern, CD3(+) , CD5(+) , CD7(+) , CD28(+) , CD80(-) , CD86(+) and human leucocyte antigen (HLA) class I+ . In contrast, HLA-DR was heterogeneously expressed. We conclude that identification and monitoring of CD4(+) Vbeta(+) clonal T cells by fluorescence-activated cell sorting with double staining is a suitable method to assess clinical responses to different therapies.