Memory CD8+T cells subjected to single-cell VDJ sequencing and mRNA sequencing, and mRNA transcripts ranked by over-expression in clonally expanded vs. non-expanded memory CD8+T cells. Slope graphs compare mRNA transcripts between expanded (n= 893) and non-expanded CD8+memory T cells (n= 3,494) from children with Wilms tumor (N = 2); expanded (n= 3,893) and non-expanded CD8+memory T cells (n= 3,000) from neuroblastoma with high TMB (above average) (N = 4); expanded (n= 430) and non-expanded CD8+memory T cells (n= 3,975) from children with neuroblastoma with low TMB (below average) (N = 5); expanded (n= 960) and non-expanded CD8+memory T cells (n= 679) from one child with a neuroblastoma with unknown TMB; expanded (n= 625) and non-expanded memory T (n= 1,415) from one child with an osteosarcoma compared with clonally expanded (n= 530) and non-expanded CD8+memory T cells (n= 275) from adult patients with colorectal cancer (CRC) (N = 12), by Zhang et al.47 (B and C) (B) As an alternative, the pediatric patient samples were compared with clonally expanded (n= 708) and non-expanded CD8+memory T cells (n= 1,515) from adult patients with renal tumors (N = 3) or lung cancer (N = 1), published by Wu et al.,38(C) or clonally expanded (n= 505) and non-expanded CD8+memory T cells (n= 180) from adult patients with non-small-cell lung cancer (N = 14), by Guo et al.48mRNA transcripts significantly overrepresented in expanded (yellow) and non-expanded cells (blue) in both adults and children are denoted. Other genes, not significantly enriched in both groups (adults or children), are colored in gray. (D) Fraction of expanded memory CD8+T cells among all memory CD8+T cells sequenced from peripheral blood of each patient in (A)–(C) shown as individual dots and groups of pediatric and adult cancer patients compared with chi-squared tests.
