Single-cell T cell receptor sequences of homo sapiens: Tumor-infiltrating leukocytes (TIL) in pancreatic ductal adenocarcinoma (PDAC) immunotherapy treated
Single-cell RNA-sequencing of tumor-infiltrating leukocytes was performed using the 10x Genomics approach. The pancreatic cancer samples were collected from surgical specimens from patients that underwent neoadjuvant immunotherapy with GVAX, cyclophosphamide, nivolumab, and 41BB agonist as monotherapies or in combinations. Treated samples were obtained from fresh non-necrotic tumor pieces resuspended in RPMI-1640 media following mechanical disruption and homogenization. Cells were filtered by a 70μm cell strainer and enriched for leukocytes by Percoll density gradient centrifugation. Cells were cryopreserved as single-cell suspensions until sequencing. Cryopreserved single cell suspensions were thawed at 37°C and underwent single-cell droplet emulsion and library preparation using the 10x Genomics Chromium single cell controller and Single Cell V(D)J Enrichment Kit ( Human T Cell) following the manufacturer's instructions. Sequencing was conducted using the HiSeq X system following manufacturer's instructions.