GSM9578551: HPV-16 specific CD4+ T cells; Homo sapiens; RNA-Seq
PBMC samples were stained with hashtag antibodies (Biolegend, custom TotalSeq anti-CD298, clone LNH-94) based on patients and/or timepoints. After two washes, PBMCs were pooled for MACS-based CD4+ T cell enrichment through negative selection (Miltenyi, 130-096-533). Isolated CD4+ T cells were incubated in 50 nM PKI solution at 37C for 20-30 min, followed by dextramer pool staining in 50 nM PKI for 20-25 min at 4C. After 2-3 washes, the cells were stained with live dye (Biolegend, 427402), CD4-BV421 (Biolegend, 317434, clone OKT4), and TCR-APC (Biolegend, 306718, clone IP26) for 10 min at 4C. The cells were then FACS sorted, washed and loaded onto Chromium Next GEM chips for single cell sequencing (10x Genomics). Library was constructed according to standard library prep manuals from 10x Genomics.