αcd14 apch7

Tetramer magnetic enrichment and single‐cell PCR were undertaken as previously described.³⁷, ⁴⁷ Briefly, PBMCs from HLA‐A*03:01⁺ individuals were FcR blocked (Miltenyi Biotech) in MACS buffer (phosphate‐buffered saline (PBS)), 0.5% bovine serum albumin (BSA; Sigma‐Aldrich); and 0.2 mm EDTA (Sigma‐Aldrich) for 15 min at 4°C. PBMCs were then stained with PE‐conjugated tetramer in MACS buffer for 1 h at room temperature, washed and labelled with anti‐PE microbeads (Miltenyi Biotech) at 4°C for 30 min. Epitope‐specific cells were enriched by passing twice over a LS magnetic column (Miltenyi Biotech) and surface stained with αCD3‐BV480 (BD Biosciences), αCD8‐PerCPCy5.5 (BD Biosciences), Live/Dead‐NIR (Life Technologies), αCD14‐APCH7 (BD Biosciences), αCD4‐APCH7 (BD Biosciences), αCD19‐APCH7 (BD Biosciences), αCD27‐BV711 (BD Biosciences), αCD45RA‐FITC (BD Biosciences), αCCR7‐PECy7 (BD Biosciences) and αCD95‐BV421 (BD Biosciences) at 4°C in the dark. Cells were resuspended in sort buffer (PBS, 0.1% BSA; Gibco, CA, USA), and tetramer^high/CD8⁺ T cells were single‐cell sorted directly into Twin‐Tech PCR plates (Eppendorf, Hamburg, Germany) on an Aria Fusion (BD Biosciences) and were stored at −80°C until used. The gating strategy is shown in Supplementary figure 1c.