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Analysis of gene expressions of mast cells in murine atopic dermatitis-like skin

ABSTRACT: Purpose: To compare gene expressions of resident and integrinβ7-positive mast cells (MCs) in atopic-dermatitis (AD) like skin. Methods: Each MC population from skin was sorted by flow cytometry, and sequencing was performed on Illumina HiSeq 4000 platform. Sequencing data in the form of FASTQ files were mapped onto the Mus musculus genome build GRCm38 using the STAR aligner (version 2.5.2b) with gene annotations obtained from GENCODE (version M22). Differential expression analysis was performed using the edgeR package (version 3.28.1) in R (version 3.6.2) and differentially expressed genes were defined as those with false discovery rate (FDR) < 0.05. Results: Principal component analysis demonstrated three distinct MC populations, and 1157 differentially expressed genes were detected. Integrinβ7-positive MC showed both mucosal type mast cell signatures and skin type mast cell signatures, and they specifically expressed migration related genes.

ORGANISM(S): Mus musculus

PROVIDER: GSE180541 | GEO | 2022/09/09

REPOSITORIES: GEO

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Project description:Although mast cells elicit proinflammatory and type I IFN responses upon VSV infection, in response to L.monocytogenes (L.m) or S. Typhimurium (S.t), such cells elicit a transcriptional program devoid of type I IFN response. Balanced induction of proinflamatory and type I interferon (IFN) responses upon activation of Toll like receptors (TLRs) determines the outcome of microbial infections and the pathogenesis of autoimmune and other inflammatory diseases. Mast cells, key components of the innate immune system, are known for their debilitating role in allergy and autoimmune syndromes. However, their potential role in anti-microbial host defenses is increasingly being acknowledged. How mast cells interact with microbes and the nature of responses triggered thereof is not well characterized. Here we show that in response to TLR activation by Gram-positive and negative bacteria or their components like LPS, unlike macrophages, mast cells elicit pro-inflammatory but not type I IFN responses. We demonstrate that in mast cells, the bound bacteria and TLR ligands remain trapped at the cell surface and do not undergo internalization - a prerequisite for type I IFN induction. Such cells could, however, elicit type I IFNs in response to vesicular stomatitis virus (VSV), which accesses the cytosolic RIG-I receptor. Although important for anti-viral immunity, a strong type I IFN response is known to contribute to pathogenesis during bacterial infection. Thus, while endowed with the capacity to elicit type I IFNs in response to viral infection, the fact that mast cells only elicit pro-inflammatory responses upon bacterial infection illustrates that mast cells, key effector cells of the innate immune system, are well adjusted for optimal anti-bacterial and anti-viral responses. Wild type control (cntr) or interferon receptor (IFNAR)-deficient mast cells (MC) or macrophages (MAC) were infected with L.m. and S.t. (MOIs 50 and 5 for MC and MAC, respectively). MC and MAC were exposed to VSV-AV2 (MOI: 2). Samples were analyzed after 6 hours. Uninfected/unstimulated cells were used as reference samples for calculating fold change in gene expression. Gene Expression levels were determined by the Affymetrix MOE 430 2.0 GeneChips. Signal Intensities were calculated using the RMA algorithm and for statistical analysis we applied GeneSpring GX 10 software suite (Agilent Technologies, Waldbronn, Germany). MultiExperiment Viewer (MEV) software version 4.4 of the Institute for Genomic Research was used for clustering algorithm data analysis and visualization.

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Analysis of gene expressions of mast cells in murine atopic dermatitis-like skin

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