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Single nuclei RNA sequencing of skin draining lymph nodes in the setting of skin inflammation and RAG1 fate mapping

ABSTRACT: Single nuclei RNA sequencing of skin draining lymph nodes in the setting of skin inflammation and RAG1 fate mapping

PROVIDER: PRJNA792191 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR17329876_1.fastq.gz	Fastqsanger.gz
	SRR17329876_2.fastq.gz	Fastqsanger.gz
	SRR17329877_1.fastq.gz	Fastqsanger.gz
	SRR17329877_2.fastq.gz	Fastqsanger.gz
	SRR17329878_1.fastq.gz	Fastqsanger.gz

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Single nuclei RNA sequencing of skin draining lymph nodes in the setting of skin inflammation and RAG1 fate mapping

Project description:Type 2 inflammation contributes to the pathology of skin diseases such as atopic dermatitis (AD) and urticaria. We previously found that AD-like inflammation induced by calcipotriol in mice is critically mediated by group 2 innate lymphoid cells (ILC2s), while T cells and B cells are dispensable given development of robust AD-like disease in RAG knockout mice. Indeed, we found that in RAG knockout mice AD-like inflammation was worse, with a proportion of ILC2s displaying increased markers of activation. The goal of this study was to characterize transcriptional and epigenomic changes in ILC populations with a history, or not, of expressing RAG1 in the setting of AD-like skin inflammation.

2024-04-28 | GSE192597 | GEO

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Project description:Topical (epicutaneous, e.c.) application of the adjuvant CpG ODN during immunization leads to a robust immune response compared to when subcutaneous (s.c.) administration. Dendritic cells are hematopoietically derived cells that are important in cross-presenting to and activating CD8 T cells. Dermal dendritic cells are one of the two major dendritic cell subsets found in the skin which mobilize from the skin to draining lymph nodes to present to T cells upon activation. Dermal dendritic cells are found in skin draining lymph nodes around 24 hours post immunization. To determine how the immune system respond differently between e.c. versus s.c. administration of CpG ODN, we evaluated changes in the skin draining lymph node environment upon the two routes of adjuvant application. Expression chemokines and chemokine receptors were assessed with real-time qPCR. To determine the changes in the skin draining lymph node environment (cytokine and cytokine receptor levels) upon immunization via real time RT-PCR.

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Cytokine and receptor expression in skin draining lymph nodes after skin immunization

Project description:Cytokine and receptor expression in skin draining lymph nodes after skin immunization

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RNA-sequencing of Vg4+Vd4+gdT cells isolated from skin-draining lymph nodes of WT and NF-kB1 deficient mice after imiquiod treatment

Project description:Purpose: The numbers of Vg4+Vd4+gdT17 cells in skin-draining lymph nodes were significantly reduced in imiqimod-treated NF-kB1-/- mice compared to those in IMQ-treated WT mice. To address the mechanism of NF-kB1-mediated proliferation of Vg4+Vd4+gdT17 cells, we performed RNA-sequencing of Vg4+Vd4+gdT cells isolated from the draining lymph nodes. Method: Mice were treated daily with 50 mg of 5% imiquimod cream or sham cream on both ears for 4 days. Single cell suspensions were separated from cervical lymph nodes. Vg4+Vd4+gdT cells were isolated by a cell sorter. N=3 each. Results: Using an optimized data analysis workflow, 552 of the transcripts showed differential expression between the WT and NF-kB1 deficient mice after imiquiod treatment, with a fold change ≥2 and p value <0.05.

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Project description:To determine the influence of primary tumors on pre-metastatic lymph nodes, we have employed whole genome microarray expression profiling as a discovery platform to identify gene signatures of stromal cells from tumor-draining lymph nodes, compared with normal lymph nodes. We subcutaneously inoculated C57BL/6 mice with the 4T1 mammary carcinoma. Two weeks later, tumor-draining lymph nodes were dissociated and stromal cells (CD45-) were sorted. Lymph nodes stromal cells from normal mice without tumor bearing were set as controls.

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Immune response genes at the mRNA vaccine delivery sites and draining lymph nodes of rhesus macaques

Project description:mRNA vaccines are emerging as a powerful vaccine platform as they are well-tolerated and scalable. Modified non-replicating mRNA encoding Influenza hemagglutinin and encapsulated in lipid nanoparticles (LNP) induced robust antibody and CD4 T cell responses after intramuscular or intradermal delivery in rhesus macaques. We investigated the local innate immune responses modulating such vaccine-induced immunity at the sites of immunization (skeletal muscle and skin) and their draining lymph nodes (LNs). Rapid mobilization of antigen presenting cells was found at the LNP/mRNA-injection sites and LNs. Dendritic cells efficiently internalized the LNPs, translated the mRNA cargo and upregulated co-stimulatory molecules. In addition, several type I interferon-inducible genes were expressed at the immunization sites and draining LNs. The innate immune activation was transient and resulted in priming of antigen-specific CD4+ T cells exclusively in the vaccine-draining LNs. Collectively, mRNA-based vaccines induce type I interferon-polarized innate immunity and antigen production by antigen presenting cells, which resulted potent vaccine-specific responses.

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