Quantitative proteomics analyses of activation states of human THP-1 macrophages

Meijer, Kees; Weening, Desireé; Vries, Marcel P. de; Priebe, Marion; Vonk, Roel J.; Roelofsen, Han

doi:10.1016/j.jprot.2015.07.013

Cited by 19 publications

(22 citation statements)

References 46 publications

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“…THP-1 cells are widely used to study monocyte/macrophage function and biology. PMA is often used to stimulate differentiation into cells that mimic human MDMs, in terms of cell morphology, macrophage surface markers, and cytokine production [ 23 ]. However, the amount of, and duration of incubation with, PMA varies widely.…”

Section: Discussionmentioning

confidence: 99%

The phorbol 12-myristate-13-acetate differentiation protocol is critical to the interaction of THP-1 macrophages with Salmonella Typhimurium

et al. 2018

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THP-1 cells differentiated with phorbol 12-myristate 13-acetate (PMA) are widely used as a model for function and biology of human macrophages. However, the conditions used for differentiation, particularly the concentration of PMA and the duration of treatment, vary widely. Here we compare several differentiation conditions and compare the ability of THP-1 macrophages to interact with the facultative intracellular pathogen Salmonella enterica serovar Typhimurium. The results show that THP-1 macrophages differentiated in high concentrations of PMA rapidly died following infection whereas those differentiated in low concentrations of PMA survived and were able to control the intracellular bacteria similar to primary human macrophages.

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Section: Discussionmentioning

confidence: 99%

The phorbol 12-myristate-13-acetate differentiation protocol is critical to the interaction of THP-1 macrophages with Salmonella Typhimurium

et al. 2018

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“…Although the functions of secreted cytokines in host response to bacterial infections have been well studied ( 27 ), other proteins, such as cathepsins ( 28 ) and specific receptors ( 29 ), can also be secreted by macrophages. There have been several attempts to identify extracellular proteins of macrophages, which included mouse RAW 264.7 macrophages ( 29 ), human U937-derived macrophages ( 30 ), mouse BMDMs ( 31 ), human primary monocyte-derived macrophages ( 32 ), human primary macrophages ( 33 ), and human THP-1-derived macrophages ( 34 ). Extracellular proteomes of cells subjected to differentiation ( 30 ) or stimulation with LPS or IL-4 ( 31 , 34 ) and influenza A virus-infected macrophages ( 33 ) were analyzed, but analysis of the extracellular proteome of human macrophages infected with bacteria has never been performed.…”

Section: Discussionmentioning

confidence: 99%

Salmonella enterica Serovar Typhimurium Alters the Extracellular Proteome of Macrophages and Leads to the Production of Proinflammatory Exosomes

et al. 2018

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Salmonella enterica serovar Typhimurium is a Gram-negative bacterium, which can invade and survive within macrophages. Pathogenic salmonellae induce the secretion of specific cytokines from these phagocytic cells and interfere with the host secretory pathways. In this study, we describe the extracellular proteome of human macrophages infected with S. Typhimurium, followed by analysis of canonical pathways of proteins isolated from the extracellular milieu. We demonstrate that some of the proteins secreted by macrophages upon S. Typhimurium infection are released via exosomes. Moreover, we show that infected macrophages produce CD63+ and CD9+ subpopulations of exosomes at 2 h postinfection. Exosomes derived from infected macrophages trigger the Toll-like receptor 4-dependent release of tumor necrosis factor alpha (TNF-α) from naive macrophages and dendritic cells, but they also stimulate secretion of such cytokines as RANTES, IL-1ra, MIP-2, CXCL1, MCP-1, sICAM-1, GM-CSF, and G-CSF. Proinflammatory effects of exosomes are partially attributed to lipopolysaccharide, which is encapsulated within exosomes. In summary, we show for the first time that proinflammatory exosomes are formed in the early phase of macrophage infection with S. Typhimurium and that they can be used to transfer cargo to naive cells, thereby leading to their stimulation.

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“…THP-1 macrophages provide several advantages over tissue resident or peripheral blood mononuclear cell (PBMC) derived macrophages: ease of access, long-term storage, faster growth rates, relative homogeneity with the same genetic background and the absence of contaminating cells [8]. Published studies have analyzed responses to polarizing stimulation by THP-1 macrophages using transcriptomic or proteomic analyses [9,10]. However, the range of phenotypic surface markers characterizing THP-1 macrophages has not been reported in detail.…”

Section: Introductionmentioning

confidence: 99%

Similarities and differences in surface receptor expression by THP-1 monocytes and differentiated macrophages polarized using seven different conditioning regimens

Forrester

Wassall

Hall

et al. 2018

Cellular Immunology

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Macrophages are key in orchestrating immune responses to micro-environmental stimuli, sensed by a complex set of surface receptors. The human cell line THP-1 has a monocytic phenotype, including the ability to differentiate into macrophages, providing a tractable, standardised surrogate for human monocyte-derived macrophages. Here we assessed the expression of 49 surface markers including Fc, complement, C-type lectin and scavenger receptors; TIMs; Siglecs; and co-stimulatory molecules by flow cytometry on both THP-1 monocytes and macrophages and following macrophage activation with seven standard conditioning/polarizing stimuli. Of the 34 surface markers detected on macrophages, 18 altered expression levels on activation. From these, expression of 9 surface markers were consistently altered by all conditioning regimens, while 9 were specific to individual polarizing stimuli. This study provides a resource for the study of macrophages and highlights that macrophage polarization states share much in common and the differences do not easily fit a simple classification system.

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Quantitative proteomics analyses of activation states of human THP-1 macrophages

Cited by 19 publications

References 46 publications

The phorbol 12-myristate-13-acetate differentiation protocol is critical to the interaction of THP-1 macrophages with Salmonella Typhimurium

The phorbol 12-myristate-13-acetate differentiation protocol is critical to the interaction of THP-1 macrophages with Salmonella Typhimurium

Salmonella enterica Serovar Typhimurium Alters the Extracellular Proteome of Macrophages and Leads to the Production of Proinflammatory Exosomes

Similarities and differences in surface receptor expression by THP-1 monocytes and differentiated macrophages polarized using seven different conditioning regimens

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