Journal: Nature Communications

Article Title: CD4 + helper T cells endow cDC1 with cancer-impeding functions in the human tumor micro-environment

doi: 10.1038/s41467-022-35615-5

Figure Lengend Snippet: CD11c − CD303 + pDC, CD11c + CD141 + cDC1, CD11c + CD1c + cDC2 and CD11c + CD14 + CD206 + moDC were flow cytometrically sorted from human PBMC as outlined in Supplementary Fig. . Cells were co-cultured with activated- or naive CD4 + T-cells overnight, then stained with antibody to β2m- conjugated to hashtag oligonucleotides (HTO) 1-8 and oligo-tagged antibody to CD3. After extensive washing steps, HTO 1-8 tagged samples were pooled in equal proportion and loaded on a 10X Genomics platform. a tSNE plots highlighting the mRNA expression profiles of pDC, cDC1, cDC2 and moDC subsets individually. Red color indicates DC co-cultured with activated (a)CD4 + T-cells. Blue color indicates DC co-cultured with naive (n)CD4 + T-cells. b GO biological process analysis using Ingenuity Pathway Analysis (IPA) using the 577 DEGs of the cDC1 “help” signature. c Dot plot depicting transcript levels and percentage of cells expressing genes related to key pathways of “antigen processing-cross presentation”, “DC maturation/migration” and “T cell differentiation/recruitment” as identified in the cDC1 “help” signature. d Heatmap revealing top 100 upregulated DEGs in the cDC1 “help” signature as derived from comparing activated CD4 + T-cell treated cDC1 (HTO2) versus naïve CD4 + T-cell treated cDC1 (HTO6).

Article Snippet: Heatmap of 114 significantly differentially expressed genes (DEGs) between DC subsets across 374 CD markers from Human Protein Atlas was generated based on log 2 FC using Qlucore Omics Explorer (version 3.7).

Techniques: Cell Culture, Staining, Expressing, Migration, Cell Differentiation, Derivative Assay

Journal: Nature Communications

Article Title: CD4 + helper T cells endow cDC1 with cancer-impeding functions in the human tumor micro-environment

doi: 10.1038/s41467-022-35615-5

Figure Lengend Snippet: To investigate the relationship between “helped” cDC1 and tumor-infiltrating DC that are conserved across multiple human solid tumor types, the cDC1 “help” signature was cross-compared with a tumor-infiltrating DC3 signature from Gerhard et al. and with tumor-infiltrating DC signatures of 8 different states that are associated with either longer or shorter overall patient survival from Luca et al. . a Venn diagram depicting number of overlapping genes between the cDC1 “help” signature and the tumor-infiltrating DC3 signature. b Heatmaps depicting the expression of tumor-infiltrating DC3 signature genes in each DC subset under “help” (aCD4 + T) or “no help” (nCD4 + T) conditions. c Heatmap depicting the expression of signature genes from the indicated tumor-infiltrating DC states (DC_S3 etc.) that are associated with longer or shorter overall survival (OS) in cDC1 under “help” (aCD4 + T) or “no help” (nCD4 + T) conditions. d Venn diagrams depicting numbers of overlapping genes between the cDC1 “help” signature and the tumor-infiltrating DC signatures of 8 different states defined in the study of Luca et al. .

Article Snippet: Heatmap of 114 significantly differentially expressed genes (DEGs) between DC subsets across 374 CD markers from Human Protein Atlas was generated based on log 2 FC using Qlucore Omics Explorer (version 3.7).

Techniques: Expressing

Journal: Nature Communications

Article Title: CD4 + helper T cells endow cDC1 with cancer-impeding functions in the human tumor micro-environment

doi: 10.1038/s41467-022-35615-5

Figure Lengend Snippet: a Heatmap depicting the expression of shared genes between cDC1 “help” signature and tumor-infiltrating DC_S3 signature (66 genes) in cDC1 under “help” (aCD4 + T) or “no help” (nCD4 + T) conditions. b – e Pearson’s correlations between ( b ) tumor-infiltrating DC3 signature , ( c ) tumor-infiltrating DC_S3 signature , ( d ) cDC1 “help” signature or ( e ) shared signature between “helped” cDC1 and tumor-infiltrating DC_S3 and defined tumor infiltrating T-cell signatures denoting activated ( a ) and effector memory (EM) CD8 + T-cells and Th1 and Th2 CD4 + T-cells within TCGA SKCM dataset ( n = 458). R, correlation coefficient. f , g Kaplan–Meier curves revealing prognostic/predictive value of tumor-infiltrating DC3 signature , tumor-infiltrating DC_S3 signature , cDC1 “help” signature or shared signature between “helped” cDC1 and tumor-infiltrating DC_S3 for ( f ) melanoma patient’s OS in TCGA SKCM cohort ( n = 458; baseline transcriptome), or ( g ) response to anti-PD-1 immunotherapy ( n = 41; baseline transcriptome). High or low metagene expression subgroups of patients were based on a median expression cut-off. p -value was calculated using Log-rank test/Mantel-Cox test (TCGA SKCM cohort) or CoxPh hazard ratios (HR), depicted as Z-scores in the anti-PD1 immunotherapy trial ( p < 0.05 is considered significant).

Article Snippet: Heatmap of 114 significantly differentially expressed genes (DEGs) between DC subsets across 374 CD markers from Human Protein Atlas was generated based on log 2 FC using Qlucore Omics Explorer (version 3.7).

Techniques: Expressing

Journal:

Article Title: Hybrids of Dendritic Cells and Tumor Cells Generated by Electrofusion Simultaneously Present Immunodominant Epitopes from Multiple Human Tumor-Associated Antigens in the Context of MHC Class I and Class II Molecules 1

doi:

Figure Lengend Snippet: DC phenotype before electrofusion. DCs from donor 4 were stained with FITC- (x-axis; FL1-H) and PE- (y-axis; FL2-H) conjugated mAbs as indicated and analyzed by FACS.

Article Snippet: After fusion, DC-888mel hybrids were detected by staining with PE-conjugated mAbs against DC cell surface markers including CD11c, CD40, CD80, CD86, CD83, and HLA-DR (BD PharMingen, San Diego, CA).

Techniques: Electrofusion, Staining

Journal:

Article Title: Hybrids of Dendritic Cells and Tumor Cells Generated by Electrofusion Simultaneously Present Immunodominant Epitopes from Multiple Human Tumor-Associated Antigens in the Context of MHC Class I and Class II Molecules 1

doi:

Figure Lengend Snippet: FACS analyses of DC-TC hybrid cells generated by electrofusion. 888 melanoma cells were labeled with CFSE (x-axis; FL1-H) and were fused with DCs from donor 4. After overnight culture, both adherent and nonadherent cell populations were stained with PE- (y-axis; FL2-H) conjugated mAbs as indicated.

Article Snippet: After fusion, DC-888mel hybrids were detected by staining with PE-conjugated mAbs against DC cell surface markers including CD11c, CD40, CD80, CD86, CD83, and HLA-DR (BD PharMingen, San Diego, CA).

Techniques: Generated, Electrofusion, Labeling, Staining