Genetic disruption of class I HLA in cutaneous T-cell lymphoma

Genetic disruption of class I HLA is a common mechanism of immune evasion and immunotherapy resistance in many other cancers but has not yet been characterized in cutaneous T-cell lymphoma (CTCL). We performed single-cell RNA sequencing (scRNA-seq) on tumor samples from patients with CTCL to demonstrate the tumor-specificity and intratumoral heterogeneity of such abnormalities. Peripheral blood mononuclear cells were isolated from whole blood using Ficoll-Paque centrifugation. Skin biopsies were dissociated into single cell suspensions using either the Tumor Dissociation Reagent (BD Biosciences) or gentleMACS Tumor Dissociation Kit (Miltenyi Biotec). After dead cell removal and CD45+ enrichment (the latter only in the case of skin biopsies), cells were analyzed using droplet-based scRNA-seq (10x Genomics).