dc marker  (Miltenyi Biotec)

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: International Journal of Molecular Sciences

Article Title: Induction of Antitumor Immunity by Exosomes Isolated from Cryopreserved Cord Blood Monocyte-Derived Dendritic Cells

doi: 10.3390/ijms21051834

Figure Lengend Snippet: DCs and their exosomes induced allogeneic T cell proliferation. ( A ) A cup-shaped morphology was observed for pulsed DC-derived exosomes by TEM. ( B ) Pulsed DC-derived exosomes expressed CD9, CD63, and CD86. ( C ) AlloT cells grew as clumps when incubated with pulsed DCs. ( D ) Carboxyfluorescein succinimidyl ester (CSFE)-stained T cells proliferated during a seven-day incubation with pulsed DCs, unpulsed DCs and exosomes isolated from DCs. Untreated T cells or T cells incubated with exosomes isolated from Exo/unpulsed DCs did not divide. ( E ) The number of T cells increased highest in the treatment with pulsed DCs, followed by Exo/pulsed DCs, and unpulsed DCs. Exo/unpulsed DCs showed no significant effect on alloT cell proliferation. Data was presented as mean ± SD in quadruplicate cultures (* p < 0.05). Exo1: pulsed DC-derived exosome sample 1; Exo2: pulsed DC-derived exosome sample 2; Exo3: pulsed DC-derived exosome sample 3. DCs: dendritic cells; pulsed DCs: A549 tumor cell lysate-pulsed DCs; unpulsed DCs: A549 tumor cell lysate-unpulsed DCs; Exo/unpulsed DCs: exosomes isolated from unpulsed DCs; Exo/pulsed DCs: exosomes isolated from pulsed DCs.

Article Snippet: The membrane was probed with antibodies (Santa Cruz Biotechnology, Dallas, TX, USA) against CD63 and CD9 (exosomal markers), CD86 (DC marker) (Beckman Counter, Marseille, France), and GAPDH followed by incubation with a horseradish peroxidase-conjugated anti-mouse antibody (Amersham TM , Freiburg, Germany).

Techniques: Derivative Assay, Incubation, Staining, Isolation