Single cell profiling of CD4+ T cells from follicular lymphoma, diffuse large B cell lymphoma and tonsilar samples

Tumor-infiltrating regulatory T cells contribute to an immunosuppressive tumor microenvironment. We applied single-cell RNA sequencing (scRNA-seq) , TCR sequencing combined with cellular indexing of transcriptomes and epitopes (CITE-seq) on CD4 T cells to decipher the heterogeneity of intratumoral Tregs in diffuse large B-cell lymphoma (DLBCL) and follicular lymphoma (FL), compared with non-malignant tonsillar tissue. Single-cell suspensions from diagnostic samples from FL and DLBCL and healthy donor tonsilar samples were stained with Alexa750 viability dye, fluorochrome-coupled-, CITE-seq- and TotalSeq-C hashtag antibodies. CD4 T cells from FL, DLBCL and tonsilar samples were isolated by fluorescence-activated cell sorting (FACS) and analyzed by scRNA-seq, TCR-seq and CITEseq.