Full Spectrum of LPS Activation in Alveolar Macrophages of Healthy Volunteers by Whole Transcriptomic Profiling
| Autor: | Bryan J. McVerry, Robert M. Tighe, Suchitra Barge, W. Michael Foster, Anastasiya Birukova, Yutong Zhao, William Horne, Rama K. Mallampalli, Prabir Ray, Anuradha Ray, Miguel Pinilla-Vera, John W. Hollingsworth, Jay K. Kolls, Janet S. Lee, Jing Zhao, Zeyu Xiong, Michael P. Donahoe |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
Lipopolysaccharides
Male 0301 basic medicine Interferon Regulatory Factor-7 medicine.medical_treatment lcsh:Medicine Biochemistry White Blood Cells Mice 0302 clinical medicine Animal Cells Interferon Medicine and Health Sciences Small interfering RNAs Alveolar Macrophages lcsh:Science Regulation of gene expression Multidisciplinary Gene Ontologies High-Throughput Nucleotide Sequencing Genomics Middle Aged Healthy Volunteers Interleukin-10 Nucleic acids Interleukin 10 Cytokine Gene Knockdown Techniques Female Ubiquitin-Specific Proteases Cellular Types Bronchoalveolar Lavage Fluid Ubiquitin Thiolesterase Research Article medicine.drug Adult Adolescent Immune Cells Immunology Biology 03 medical and health sciences Extraction techniques Gene Types Endopeptidases Macrophages Alveolar Genetics medicine Animals Humans Non-coding RNA Aged Blood Cells Macrophages lcsh:R Biology and Life Sciences Proteins Computational Biology Cell Biology Macrophage Activation Genome Analysis Molecular biology RNA extraction Gene regulation Research and analysis methods RAW 264.7 Cells 030104 developmental biology Gene Expression Regulation TRIF TLR4 RNA Regulator Genes IRF7 lcsh:Q Interferons Gene expression Transcriptome 030215 immunology Interferon regulatory factors |
| Zdroj: | PLoS ONE, Vol 11, Iss 7, p e0159329 (2016) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | Despite recent advances in understanding macrophage activation, little is known regarding how human alveolar macrophages in health calibrate its transcriptional response to canonical TLR4 activation. In this study, we examined the full spectrum of LPS activation and determined whether the transcriptomic profile of human alveolar macrophages is distinguished by a TIR-domain-containing adapter-inducing interferon-β (TRIF)-dominant type I interferon signature. Bronchoalveolar lavage macrophages were obtained from healthy volunteers, stimulated in the presence or absence of ultrapure LPS in vitro, and whole transcriptomic profiling was performed by RNA sequencing (RNA-Seq). LPS induced a robust type I interferon transcriptional response and Ingenuity Pathway Analysis predicted interferon regulatory factor (IRF)7 as the top upstream regulator of 89 known gene targets. Ubiquitin-specific peptidase (USP)-18, a negative regulator of interferon α/β responses, was among the top up-regulated genes in addition to IL10 and USP41, a novel gene with no known biological function but with high sequence homology to USP18. We determined whether IRF-7 and USP-18 can influence downstream macrophage effector cytokine production such as IL-10. We show that IRF-7 siRNA knockdown enhanced LPS-induced IL-10 production in human monocyte-derived macrophages, and USP-18 overexpression attenuated LPS-induced production of IL-10 in RAW264.7 cells. Quantitative PCR confirmed upregulation of USP18, USP41, IL10, and IRF7. An independent cohort confirmed LPS induction of USP41 and IL10 genes. These results suggest that IRF-7 and predicted downstream target USP18, both elements of a type I interferon gene signature identified by RNA-Seq, may serve to fine-tune early cytokine response by calibrating IL-10 production in human alveolar macrophages. |
| Databáze: | OpenAIRE |
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