An integrated reagent and multimodal analysis workflow to enrich and characterize peptide-specific CD8+T cells

Abstract Immune responses to disease are complex and the only way understand such a response is consider behavior of multiple different cell types across both innate adaptive branches immune system. We developed Barcode Enabled Antigen Mapping (BEAM) technology enable easy enrichment antigen-specific cells. This fully integrated with 10x Genomics Chromium Single Cell Profiling Solution allow multimodal evaluation cells including full-length, paired sequences T B receptor genes while simultaneously screening their specificity for set genes. Our TCR workflow (BEAM-T) identifies CD8+T demonstrate assay’s performance spike-in experiments using various percentages HLA-A*02:01-restricted peptide-stimulated (targeted mixture being 40% cytomegalovirus peptide, 10% flu peptide 0–5% anti-SARS-CoV-2 cells) into background human PBMCs. Cells were labeled panel five viral pathogen peptide-loaded multimers, plus HLA-A*02:01 negative control assess antigen then sorted CD8+PE+ demonstrated that major VDJ-T clonotypes identified SARS-CoV2 retained, albeit roughly proportionally fewer cells, as percentage spiked decreased. further feature robustness protocol sample models H2Kb-restricted OT-1 transgenic mouse splenocytes, well healthy donor PBMCs dissociated tumor utility an reagent-to-data analysis offering range samples.