Improved Sézary cell detection and novel insights into immunophenotypic and molecular heterogeneity in Sézary syndrome

• EuroFlow standardized multiparameter flow cytometry improves identification, quantitation, and characterization of circulating Sézary cells.
• Immunophenotypically distinct Sézary subsets are clonally related and share a transcriptomic signature indicating impaired T-cell function.

Sézary syndrome (SS) is an aggressive leukemic form of Cutaneous T-cell Lymphoma with neoplastic CD4+ T cells present in skin, lymph nodes, and blood. Despite advances in therapy, prognosis remains poor with a 5-year overall survival of 30%. The immunophenotype of Sézary cells is diverse, which hampers efficient diagnosis, sensitive disease monitoring, and accurate assessment of treatment response. Comprehensive immunophenotypic profiling of Sézary cells with an in-depth analysis of maturation and functional subsets has not been performed thus far. We immunophenotypically profiled 24 SS patients employing standardized and sensitive EuroFlow-based multiparameter flow cytometry (MFC). We accurately identified and quantified Sézary cells in blood and performed an in-depth assessment of their phenotypic characteristics in comparison with their normal counterparts in the blood CD4+ T-cell compartment. We observed inter-and intra-patient heterogeneity and phenotypic changes over time. Sézary cells exhibited phenotypes corresponding with classical and non-classical T helper subsets with different maturation phenotypes.