Fig. 4: scRNA-seq analysis of individual CARMz T (sCARMz T) and CARMz T from a mixture of CARMz T and CARP T cells (mCARMz T) after coculture with HeLa-GL cells. | Nature Communications

Fig. 4: scRNA-seq analysis of individual CARMz T (sCARMz T) and CARMz T from a mixture of CARMz T and CARP T cells (mCARMz T) after coculture with HeLa-GL cells.

From: Co-expression of a PD-L1-specific chimeric switch receptor augments the efficacy and persistence of CAR T cells via the CD70-CD27 axis

Fig. 4

a–h Single cell RNA-seq strategy: Individual CARMz T (sCARMz T) and CARMz T from a combination of CARMz T and CARP T cells (mCARMz T) were separated by flow cytometry sorting based on their GFP tag post-coculture with HeLa-GL cells, and then processed for scRNA-seq. b The UMAP projection of single CARMz T from sCARMz T and mCARMz T cells. C1-Tua: non-activated T cells, C2-Tef8: CD8+ effector T cells, C3-Tie8: partially differentiated CD8+ effector T cells, C4-Tcm8: CD8+ central memory T cells, C5-acTcm8: activated CD8+ central memory T cells, C6-Tef4: CD4+ effector T cells, C7-Tie4: partially differentiated CD4+ effector T cells, C8-Tcm4: CD4+ central memory T cells. Each dot corresponds to a single cell. c Single cells from sCARMz T (red) and mCARMz T (blue) clusters in distinct regions of the UMAP space. d Single-cell transcript levels of CD8, CD4, CCL5, TNFRSF4, CCR7, and CD27 illustrated in UMAP plots. e Dot plot of selected DEGs expressed in each cluster. f, g GSEA illustrating the enrichment of genes defined as β€œCD8 stem cell memory vs. naive CD8 T cell UP” in cluster Tcm8 and genes defined as β€œeffector memory vs. central memory CD4 T cell UP” in cluster Tcm4. NES (Normalized Enrichment Scores) and p adjust are indicated. h Bar graphs summarizing the number of sCARMz T and mCARMz T cells in each cluster. i Representative phenotype of separated CARMz T (sCARMz T), CARMz T from a mixture of CARMz T and CARP T (mCARMz T), and control CAR19z T cells (gated on CD8+GFP+ cells and CD8βˆ’GFP+ cells) post-coculture with HeLa cells for 36 h. j Proportion of Tcm (central memory T cells, CD45RO+CCR7+) in sCARMz T, mCARMz T and control CAR19z T cells in (i). Data are presented as mean ± SD (N = 3 biological samples). p Values were calculated by two-way ANOVA with Tukey’s multiple comparisons test (mCARMz T vs. sCARMz T = CD8+: 3.913Eβˆ’05, CD4+: 2.268Eβˆ’04). ***p < 0.001.

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