T cells in B16 tumor model with ACT and checkpoint blockade - scRNA-seq, CITE-seq and TCRab [i35]

This study is part of the immgenT Open Source Project, specifically from IGT35, which focuses on characterizing T cells in the B16.gp33 melanoma model following adoptive cell therapy (ACT) and immune checkpoint blockade. T cells were isolated from tumors and draining inguinal lymph nodes of C57BL/6J male mice (7–8 weeks old) subjected to subcutaneous implantation of B16.gp33 melanoma cells, followed by adoptive cell transfer treatment with intravenous transfer of P14 and pmel T cells, and treatment with either isotype control antibodies or a combination of anti-PD-1 and anti-4-1BB antibodies. Samples were collected at day 14 post post-tumor implantation (1 day post-final antibody treatment). The study targeted all CD3+ T cell populations, including transgenic P14 and pmel T cells Overall design: T cells were isolated from tumors and draining inguinal lymph node samples using mechanical dissociation or collagenase I digestion. Sorted CD3+ T cell populations, including transgenic P14 and pmel T cells were profiled using single-cell RNA sequencing (10x Genomics), combined with CITE-seq (TotalSeq C) and paired TCR a/ß sequencing. Samples from multiple mice were pooled using hashtag oligonucleotides for multiplexing.