Journal: Nature Communications
Article Title: The CD4 + T cell population partners with Tpex CD8 + T cells to mediate antitumor immunity in the tumor microenvironment
doi: 10.1038/s41467-026-71161-0
Figure Lengend Snippet: a UMAP plots of CD3D + CD4 + cells generated with integrated gene expression data from single-cell RNA sequencing (scRNA-seq) of peripheral blood (PB) and tumor-infiltrating lymphocytes (TILs) derived from 8 stage I-II non-small cell lung cancer patients. All 79,130 CD3D + CD4 + cells (65,572 PB-CD3D + CD4 + cells and 13,558 TIL-CD3D + CD4 + cells) were categorized into 27 clusters, as illustrated in the UMAP plot. b The expression of SELL in the UMAP plot is indicated. The area outlined by the red dotted line represents clusters composed of effector T cells with downregulated expression of SELL . c On the basis of T-cell receptor (TCR) repertoire analysis using scTCR-seq, cells belonging to TCR clonotypes with two or more cells detected were mapped as pink dots on the UMAP plot. Although Foxp3 + Tregs in TILs included multicellular TCR clonotype cells, most effector multicellular TCR clonotype cells belong to clusters in which SELL expression was downregulated (outlined by a red dotted line). d Expression of FoxP3 in the UMAP plot is indicated. e Expression patterns of the 58 signature genes from sorted Th1, Th7R, and Th17 cells, and clusters PB-13 + 26, 18 + 21, and 14 from unsupervised clustering of surgical samples are shown as a heatmap. Predicted probabilities and cosine similarity index with sorted Th are indicated. f Distribution of cells with the same TCR clonotype found in the TIL-13, TIL-14, TIL-18, and TIL-21 clusters in both the TIL and the PB clusters. Each color represents cluster identity as defined in the UMAP plot (Fig. 1a). The TCR clonotypes found in the TIL-13 cluster were found almost exclusively in cluster 13 in TILs or PB. However, TIL-21 clonotypes were shared with the cluster 18 in TILs. g The number of cells in each TIL and PB cluster, those belonging to the 13, 14, 18, 21, and 26 clusters, are plotted in a color-coded stacked bar graph divided by five levels of clonal expansion. The PB-18 cluster had very few multicellular TCR clonotypes, whereas the TIL-18 cluster contained substantially expanded TCR clonotypes. h UMAP plot based on the WNN method using nuclear mRNA expression (nRNA) and chromatin accessibility (ATAC) profiles. A total of 10,699 nuclei were classified into 27 distinct clusters. i Annotation of Th1, Th7R, Th17, Treg and other cell type were visualized in a UMAP plot. j Nuclear mRNA expression patterns of 58 signature genes. The results of the cosine similarity analysis of the gene expression patterns in each cluster compared with the gene expression of the sorted Th cells are also presented. k Gene activity of 54 of 58 signature genes were shown as heatmap. The results of the cosine similarity analysis of the gene expression patterns in each cluster compared with those in the sorted Th cells are also presented.
Article Snippet: All single-cell RNA-seq and scTCR-seq libraries were constructed using the Chromium controller and Chromium Single-Cell Immune Profiling v2 kit (10x Genomics Inc., Pleasanton, CA, USA) to analyze gene expression differences and T-cell receptor (TCR) repertoires among T-cell subpopulations.
Techniques: Generated, Gene Expression, Single Cell, RNA Sequencing, Derivative Assay, Expressing, Activity Assay