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. 2022 May 24;25(2):263–276. doi: 10.1093/neuonc/noac138

Fig. 2.

Fig. 2

HGG-infiltrating dendritic cells exhibit limited antigen presentation via MHC class I. (A) Experimental overview. GL261-OVA or wildtype control cells were implanted in n = 12 C57BL6/J mice. 28 days post implantation, tumors and meninges were isolated and subjected to flow cytometry of SIINFEKL-reactive T cells and SIINFEKL-presenting DCs (CD11c+/MHC-II +). (B) Representative flow cytometry gating of H-2Kb-SIINFEKL-presenting DCs and SIINFEKL-dextramer-positive T cells isolated from syngeneic gliomas. Left column: GL261-Ova; Right column: GL261-WT. (C) left: Quantification of H-2Kb-SIINFEKL-presenting DCs and SIINFEKL-dextramer-positive T cells in meninges and glioma samples. n = 6 biological replicates per group. P-values are derived from paired student’s t-tests. right: inter-individual analysis of matched SIINFEKL-presenting DCs and SIINFEKL-dextramer-positive T cells from (left). Mouse IDs indicated. (D–F) Single-cell RNA-Seq of CD45+ cells from IDHmut and IDHwt GL261 mouse experimental gliomas. t-SNE maps are color-coded for (D) the genotype of the glioma and (E) the identified cell types based on marker gene expression. Median expression of cDC1 marker gene signature (Itgae, Xcr1, Sept3, Gcsam, Clec9a, Pianp, Ffar4, A503099J19Rik, Plpp1, Cxx1a, Ifi205, Cyp8b1, Tlr3, Snx2) and cDC2 signature (CD209a, Tnfsf9, Tnip3, Kcne3) shown in (D). Analysis was conducted on n = 7910 cells from IDH-mutated experimental glioma and n = 8835 cells from IDH-wildtype experimental glioma. (F) Heatmap representation of differentially regulated genes across all clusters. Clusters 5, 9, 11, 15, 18 represent cells on the monocyte-to-DC trajectory. Relative gene expression shown.