title: Establishing a Proteomics-Based Monocyte Assay To Assess Differential Innate Immune Activation Responses.
abstract: Innate immune cells are complex systems that can be simultaneously activated in a variety of ways. Common methods currently used to estimate the response of innate immune cells to stimuli are usually biased toward a single mode of activation. The aim of this study was to assess the possibility of designing an assay based on unbiased proteome analysis that would be capable of predicting the complex response of the innate immune system to various challenges. Monocytes were used as representative cells of the innate immune system. The underlying hypothesis was that their proteome response to different activating molecules would reflect the immunogenicity of these molecules. To identify the main modes of response, we treated the human monocytic THP-1 cell line with nine different stimuli. Differentiation and activation were determined to be the two major modes of monocyte response, with PMA causing the strongest differentiation and Pam3CSK4 causing the strongest proinflammatory activation. The established assay was applied to characterize the monocyte response to epidermal growth factor peptide containing isoaspartate, which induced differentiation but not proinflammatory activation. Because of its versatility, robustness, and specificity, this new assay is likely to find a niche among the more established immunological methods.
authors.forename: Nataliya K
authors.initials: NK
authors.lastname: Tarasova
authors.forename: A Jimmy
authors.initials: AJ
authors.lastname: Ytterberg
authors.forename: Karin
authors.initials: K
authors.lastname: Lundberg
authors.affiliation: Department of Clinical Neuroscience, Centre for Molecular Medicine, Karolinska Hospital, Karolinska Institutet , SE 17176 Stockholm, Sweden.
authors.forename: Xing-Mei
authors.initials: XM
authors.lastname: Zhang
authors.affiliation: Department of Clinical Neuroscience, Centre for Molecular Medicine, Karolinska Hospital, Karolinska Institutet , SE 17176 Stockholm, Sweden.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Harris
authors.forename: Roman A
authors.initials: RA
authors.lastname: Zubarev
mesh_terms: D002460:Cell Line; D004815:Epidermal Growth Factor / Q000494:pharmacology; D006801:Humans; D007113:Immunity, Innate; D055666:Lipopeptides / Q000494:pharmacology; D015166:Monitoring, Immunologic / Q000379:methods* / Q000592:standards; D009000:Monocytes / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D020543:Proteome / Q000187:drug effects* / Q000276:immunology; D040901:Proteomics / Q000379:methods*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
chemical_list: D055666:Lipopeptides; C508075:Pam(3)CSK(4) peptide; D020543:Proteome; D004815:Epidermal Growth Factor; D013755:Tetradecanoylphorbol Acetate
keywords: THP-1 cell line; TLR ligands; activation; differentiation; label-free proteomics