title: Full Spectrum of LPS Activation in Alveolar Macrophages of Healthy Volunteers by Whole Transcriptomic Profiling.
abstract: 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.
authors.affiliation: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Miguel
authors.initials: M
authors.lastname: Pinilla-Vera
authors.affiliation: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Zeyu
authors.initials: Z
authors.lastname: Xiong
authors.affiliation: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.|Vascular Medicine Institute, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Yutong
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.|Vascular Medicine Institute, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Jing
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Michael P
authors.initials: MP
authors.lastname: Donahoe
authors.affiliation: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Suchitra
authors.initials: S
authors.lastname: Barge
authors.affiliation: Department of Pediatrics, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: William T
authors.initials: WT
authors.lastname: Horne
authors.affiliation: Department of Pediatrics, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Jay K
authors.initials: JK
authors.lastname: Kolls
authors.affiliation: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Bryan J
authors.initials: BJ
authors.lastname: McVerry
authors.affiliation: Department of Medicine, Duke University, Durham, North Carolina, United States of America.
authors.forename: Anastasiya
authors.initials: A
authors.lastname: Birukova
authors.affiliation: Department of Medicine, Duke University, Durham, North Carolina, United States of America.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Tighe
authors.affiliation: Department of Medicine, Duke University, Durham, North Carolina, United States of America.
authors.forename: W Michael
authors.initials: WM
authors.lastname: Foster
authors.affiliation: Department of Medicine, Duke University, Durham, North Carolina, United States of America.|Department of Medicine, Division of Pulmonary and Critical Care Medicine, Ohio State University, Columbus, Ohio, United States of America.
authors.forename: John
authors.initials: J
authors.lastname: Hollingsworth
authors.affiliation: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Anuradha
authors.initials: A
authors.lastname: Ray
authors.affiliation: The Medical Specialty Service Line, Veterans Affairs Pittsburgh Healthcare System, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Rama
authors.initials: R
authors.lastname: Mallampalli
authors.affiliation: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Prabir
authors.initials: P
authors.lastname: Ray
authors.affiliation: Department of Medicine, Division of Pulmonary, Allergy, and Critical Care Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.|Vascular Medicine Institute, University of Pittsburgh, Pittsburgh, Pennsylvania, United States of America.
authors.forename: Janet S
authors.initials: JS
authors.lastname: Lee
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology; D010450:Endopeptidases / Q000235:genetics*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D055785:Gene Knockdown Techniques; D064368:Healthy Volunteers; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D050839:Interferon Regulatory Factor-7 / Q000037:antagonists & inhibitors / Q000235:genetics*; D016753:Interleukin-10 / Q000235:genetics*; D008070:Lipopolysaccharides / Q000008:administration & dosage / Q000378:metabolism; D008262:Macrophage Activation / Q000187:drug effects / Q000235:genetics; D016676:Macrophages, Alveolar / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008875:Middle Aged; D000067996:RAW 264.7 Cells; D059467:Transcriptome / Q000235:genetics; D043222:Ubiquitin Thiolesterase; D064570:Ubiquitin-Specific Proteases / Q000235:genetics*
chemical_list: C508609:IL10 protein, human; C494234:IRF7 protein, human; D050839:Interferon Regulatory Factor-7; D008070:Lipopolysaccharides; D016753:Interleukin-10; D010450:Endopeptidases; C485560:USP18 protein, human; C579217:USP41 protein, human; D043222:Ubiquitin Thiolesterase; D064570:Ubiquitin-Specific Proteases