scRNA-seq and CITE-seq of immune cells at day 3 after ACT using Pmel TCR-transgenic CD39⁺CD69⁺

Concurrent neoantigen-vaccination enhances the antitumor activity of ACT-products dominated by dysfunctional T cells (Tdys cells). To understand if concurrent vaccination influences early changes in the immune landscape within B16^KVP TME following T_dys-ACT into lymphodepleted hosts that led to improved antitumor response. To this end, 3 days after ACT, we performed single-cell RNA sequencing (scRNA-seq) along with cellular indexing of transcriptomes and epitopes by sequencing (CITE-seq) by “cell hashing” of 4 groups of CD45⁺ tumor-infiltrating immune cells: those derived from untreated mice, mice receiving VACV^KVP (vaccine) only, and mice receiving T_dys-ACT (using CD39⁺CD69⁺FACS-sorted cells) with and without vaccination.