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Hiltonol and pRNA stimuli induce an immune activating profile in the transcriptomics of cDC1s [RNA-seq]

ABSTRACT: Hiltonol and pRNA stimuli induce an immune activating profile in the transcriptomics of cDC1s [RNA-seq]

PROVIDER: PRJNA1001363 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR25497610_1.fastq.gz	Fastqsanger.gz
	SRR25497610_2.fastq.gz	Fastqsanger.gz
	SRR25497611_1.fastq.gz	Fastqsanger.gz
	SRR25497611_2.fastq.gz	Fastqsanger.gz
	SRR25497612_1.fastq.gz	Fastqsanger.gz

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Hiltonol and pRNA stimuli induce an immune activating profile in the transcriptomics of cDC1s

Project description:Hiltonol and pRNA stimuli induce an immune activating profile in the transcriptomics of cDC1s

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Hiltonol and pRNA stimuli induce an immune activating profile in the transcriptomics of cDC1s

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Hiltonol and pRNA stimuli induce an immune activating profile in the transcriptomics of cDC1s [Affy]

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Hiltonol and pRNA stimuli induce an immune activating profile in the transcriptomics of cDC1s

Project description:8 ligand), and compared these samples to unstimulated counterparts. Both methods, RNA-seq and microarray, gave very similar results and showed that Hiltonol and pRNA lead to almost identical changes in the transcriptome of cDC1 mDCs. A gene ontology (GO) term analysis suggested that these changes were mainly related to activation and maturation pathways, including induction of type I IFN and IL-12 transcription, while pathways related to adverse effects or cell damage were not strongly affected. The combination of both reagents in the DC cultures gave a very similar result as compared to either stimulus alone, suggesting no synergistic effect. Together, our results indicate that both stimuli are potent clinical grade adjuvants with comparable effects to mature cDC1 mDCs within an immunogenic profile in short-term cultures suitable to be used in immunotherapy.

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Hiltonol and pRNA stimuli induce an immune activating profile in the transcriptomics of cDC1s [Affy]

Project description:Stimulation of dendritic cells (DC) is considered critical in cancer immunotherapy. BATF-3–dependent subsets, that express in humans BDCA-3 (CD141), promote CD8 T-cell cross-priming against tumor antigens. Here, we evaluate two clinical grade stimuli for peripheral blood cDC1 myeloid dendritic cells (mDCs), a rare DC subset that is currently being explored for use in immunotherapy. In contrast to routine evaluation methods, which focus on predefined maturation markers on the surface or factors released from the activated cells, we applied an unbiased transcriptome-based method using both RNA-sequencing (RNA-seq) and microarrays. Specifically, we analyzed the mRNA of cDC1 mDCs from five human donors upon activation with two clinical-grade adjuvants, Hiltonol (poly IC, a TLR3 ligand, called Stimulus 1 in the description below) and protamine RNA (pRNA, aTLR7/8 ligand, Stimulus 2), and compared these samples to unstimulated counterparts. Both methods, RNA-seq and microarray, gave very similar results and showed that Hiltonol and pRNA lead to almost identical changes in the transcriptome of cDC1 mDCs. A gene ontology (GO) term analysis suggested that these changes were mainly related to activation and maturation pathways, including induction of type I IFN and IL-12 transcription, while pathways related to adverse effects or cell damage were not strongly affected. The combination of both reagents in the DC cultures gave a very similar result as compared to either stimulus alone, suggesting no synergistic effect. Together, our results indicate that both stimuli are potent clinical grade adjuvants with comparable effects to mature cDC1 mDCs within an immunogenic profile in short-term cultures suitable to be used in immunotherapy.

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Hiltonol and pRNA stimuli induce an immune activating profile in the transcriptomics of cDC1s [RNA-seq]

Project description:Stimulation of dendritic cells (DC) is considered critical in cancer immunotherapy. BATF-3–dependent subsets, that express in humans BDCA-3 (CD141), promote CD8 T-cell cross-priming against tumor antigens. Here, we evaluate two clinical grade stimuli for peripheral blood cDC1 myeloid dendritic cells (mDCs), a rare DC subset that is currently being explored for use in immunotherapy. In contrast to routine evaluation methods, which focus on predefined maturation markers on the surface or factors released from the activated cells, we applied an unbiased transcriptome-based method using both RNA-sequencing (RNA-seq) and microarrays. Specifically, we analyzed the mRNA of cDC1 mDCs from five human donors upon activation with two clinical-grade adjuvants, Hiltonol (poly IC, a TLR3 ligand, called "Stimulus 1" in the description below) and protamine RNA (pRNA, aTLR7/8 ligand, "Stimulus 2"), and compared these samples to unstimulated counterparts. Both methods, RNA-seq and microarray, gave very similar results and showed that Hiltonol and pRNA lead to almost identical changes in the transcriptome of cDC1 mDCs. A gene ontology (GO) term analysis suggested that these changes were mainly related to activation and maturation pathways, including induction of type I IFN and IL-12 transcription, while pathways related to adverse effects or cell damage were not strongly affected. The combination of both reagents in the DC cultures gave a very similar result as compared to either stimulus alone, suggesting no synergistic effect. Together, our results indicate that both stimuli are potent clinical grade adjuvants with comparable effects to mature cDC1 mDCs within an immunogenic profile in short-term cultures suitable to be used in immunotherapy.

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Erythropoietin receptor on cDC1s dictates immune tolerance

Project description:Immune tolerance is an active state of unresponsiveness of the immune system to antigens (Ags) that have the potential to induce an immune response1. Such tolerance is beneficial in avoiding autoimmunity and rejection of organ transplants, but detrimental in cancer2. Type 1 conventional dendritic cells (cDC1s) are unique in their efferocytosis3 and cross-presenting abilities4, resulting in T cell-mediated immunity5 or tolerance6-10. However, the mechanisms underlying the tolerogenic function of cDC1s remain largely unknown. Here, we show that the erythropoietin receptor (EpoR) acts as a critical switch that determines the tolerogenic state of cDC1s and the threshold of Ag-specific T cell responses. In total lymphoid irradiation-induced allograft tolerance11,12, EpoR+ cDC1s induce donor-specific CD4+FoxP3+ regulatory T cells (Tregs), and conditional knockout of EpoR in cDC1s abrogates Ag-specific Treg induction, resulting in allograft rejection. In the steady state, EpoR+ migratory cDC1s control Ag-specific Treg induction in peripheral lymph nodes (LNs). In cancer, EpoR is expressed on both tumor Ag-carrying, tumor-infiltrating cDC1s, as well as migratory cDC1s in tumor-draining LNs (tdLNs). Loss of EpoR from cDC1s leads to tumor reduction by enhancing tumor Ag-specific CD8+ T cell priming and generating more precursor exhausted T cells13 in tdLNs, preserving CD8+ T cell precursor-like features and effector function, and reducing Tregs in the tumor. Thus, targeting EpoR on cDC1s to induce or inhibit immune tolerance should enable new ways to treat a variety of diseases.

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Expression data from WT cDC1s, Zeb2 KO cDC1s, Nfil3 Zeb2 DKO cDC1s and Zeb2 Id2 DKO cDC1s

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Expression data from WT cDC1s, Zeb2 KO cDC1s, Nfil3 Zeb2 DKO cDC1s and Zeb2 Id2 DKO cDC1s

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