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

Starr, Tregei; Bauler, Timothy J.; Malik-Kale, Preeti; Steele‐Mortimer, Olivia

doi:10.1371/journal.pone.0193601

Cited by 131 publications

(127 citation statements)

References 30 publications

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“…Light microscopy analysis revealed changes in PMA induced morphology, including increased cellular adhesion and spread morphology. In concordance with the results observed by Starr et al (28), the changes in cell morphology were dependent on the concentration and the duration of incubation, with cells treated with PMA and rested for two days showing a significant increase in cytoplasmic volume with increased adherence ( Figure 1A ). The differentiation was more pronounced in Condition A in comparison with the conditions B and C. Additionally, flow cytometry analyses indicated an increase in side scatter (SSC), in condition A with respect to conditions B and C. The cells treated with PMA but not rested (Condition C) closely resembled the undifferentiated THP-1 cells in regard to these properties ( Figure 1B ).…”

Section: Resultssupporting

confidence: 91%

“…However, to be used as an in vitro model mimicking human macrophages, THP-1 cells have to be differentiated, and several protocols have been tested (7, 9, 29, 44). Among these, PMA is most often employed to induce differentiation, with almost similar phenotypes reported in terms of cell morphology, expression of macrophage surface markers, and cytokine production (28, 45). However, the amount and duration of incubation with PMA vary widely across the literature.…”

Section: Discussionmentioning

confidence: 99%

“…However, the amount and duration of incubation with PMA vary widely across the literature. Additionally, studies demonstrate altered sensitivity, as well as undesirable gene regulation in PMA-differentiated macrophages that may contribute to their differential response to secondary stimuli (12, 28). Although this may likely be the case, one inherent limitation of most of these studies is that their focus has primarily been on characterizing either the transcriptional profile or surface receptor expression (11, 30).…”

Section: Discussionmentioning

confidence: 99%

“…Depth of 100. Kinome trees were drawn using KinMap (28) (http://kinhub.org/kinmap). Gene lists for functions such as phagocytosis, reactive oxygen species, and inflammasome complex were obtained from the Molecular Signatures Database (MSigDB, v7.0, https://www.gsea-msigdb.org/gsea/msigdb) (29).…”

Section: Methodsmentioning

confidence: 99%

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Dose-dependent phorbol 12-myristate-13-acetate-mediated monocyte-to-macrophage differentiation induces unique proteomic signatures in THP-1 cells

Pinto

Kim

Subbannayya

et al. 2020

Preprint

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1, 2). Monocytes in the blood circulation migrate to the site of infection/inflammation and 48 differentiate into macrophages for effective host defense, tissue remodeling, and repair (3). 49Furthermore, macrophages exhibit a high level of plasticity, depending on their local 50 microenvironment, specialized functions and varied phenotype acquired (1, 4). 51Several models are employed to study the mechanisms of immune-modulation in monocytes 52 and macrophages during inflammatory diseases. The most frequently used include primary 53 peripheral blood mononuclear cells (PBMCs) and monocyte cell lines. However, due to donor-54 to-donor variations and technical disparities involved in handling of PBMCs in vitro, the 55 human leukemia monocytic cell line, THP-1, is widely accepted and used as a 56 monocyte/macrophage model (5, 6). Several studies have indicated that THP-1 cells can be 57 differentiated into macrophage-like cells using phorbol-12-myristate-13-acetate (PMA), 58 which markedly resembles PBMC monocyte-derived macrophages (MDMs) in cytokine 59 production, metabolic and morphological properties, including differential expression of 60 macrophage surface markers such as CD14, CD11b (ITGAM) and scavenger receptors-CD163, 61 MSR1, and SCARB2 (5, 7-10). Nonetheless, depending on the parameters of the differentiation 62 protocol employed, such as the concentration (ranging from 5 to 400 ng/mL), and duration of 63 incubation (1 to 5 days) with PMA; the degree of differentiation and the functional changes 64 may vary significantly (5,(11)(12)(13)(14)(15). 65At the molecular level, multiple proteins including growth factors, antigenic markers, 66 chemokine-receptors, cytokines, and cell adhesion molecules, are known to govern and reflect 67 underlying monocyte-macrophage differentiation processes (16)(17)(18)(19)(20). However, the effect of 68 various differentiation protocols on the cellular proteome and intracellular signaling networks 69 4 during monocyte-to-macrophage differentiation remains poorly understood. Hence, it is crucial 70 to determine the most suitable differentiation conditions when using these cells as model 71 system, as this can significantly impact their response to various innate immune stimuli. 72Quantitative high-resolution mass spectrometry-based proteomic approaches have been widely 73 employed to investigate the proteomes of monocytes and macrophages as well as altered 74 cellular proteomes and complex cellular/biological mechanisms in several biological 75 conditions (21-23). However, to date, no studies have directly compared the proteome changes 76 of the PMA-mediated differentiation process. 77In the present study, we evaluate the effect of three PMA-based differentiation protocols on 78 the changes in the proteome profiles upon THP-1 differentiation using stable isotope dimethyl 79 labeling quantitative proteomics. We demonstrate that various differentiation conditions such 80 as concentration and incubation time, prior to any stimuli, are critical consideration factors for 81 hetero...

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Section: Resultssupporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 2 more Smart Citations

Dose-dependent phorbol 12-myristate-13-acetate-mediated monocyte-to-macrophage differentiation induces unique proteomic signatures in THP-1 cells

Pinto

Kim

Subbannayya

et al. 2020

Preprint

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“…Differentiated cells exhibited macrophage-like phenotypes, which were characterized by their morphology and increased cell surface expression of CD11b by flow cytometry analysis (Fig. S1) as described in a previous study (10). THP-1 macrophages were used in the subsequent experiments.…”

Section: Methodsmentioning

confidence: 82%

Effects of nanoparticle‑mediated delivery of pitavastatin on atherosclerotic plaques in ApoE‑knockout mice and THP‑1‑derived macrophages

et al. 2020

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The treatment of atherosclerosis remains complex. Pitavastatin serves an important role in the prevention and treatment of atherosclerosis. The present study aimed to investigate the effects of nanoparticle (NP)-mediated delivery of pitavastatin into atherosclerotic plaques as a novel treatment method for atherosclerosis. The results of the present study demonstrated that pitavastatin-NP was more effective in attenuating the size of atherosclerotic plaques and enhancing the stability of plaques in vitro compared with pitavastatin alone. In an apolipoprotein E (ApoE)-knockout mouse model of atherosclerosis, a single intravenous injection of fluorescein isothiocyanate-NP resulted in the delivery of NP into atherosclerotic plaques for up to 7 days post-injection. In ApoE-knockout mice and THP-1-derived macrophages, pitavastatin-NP attenuated the development of atherosclerosis, which was associated with regulating lipid metabolism, and inhibited the secretion of inflammatory markers compared with pitavastatin alone. Additionally, the treatment advantages of pitavastatin-NP were independent of lipid lowering. The results demonstrated that pitavastatin-NP administration was more effective in attenuating the development of atherosclerotic plaques compared with systemic administration of pitavastatin.

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Engineering Living Immunotherapeutic Agents for Improved Cancer Treatment

Gwisai,

Günther,

Mirkhani

et al. 2023

Advanced Therapeutics

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Bacteria‐based agents are emerging as promising tools for cancer therapy due to their ability to actively target tumors, trigger localized inflammation, and induce tumor regression. There has been growing interest in using bacteria that are responsive to external cues, such as magnetic fields, to facilitate the formation of robust colonies in tumors and achieve the threshold for clinical efficacy. Several studies have demonstrated the potential of innately magnetically responsive bacteria, known as magnetotactic bacteria (MTB), as steerable agents. However, their immunostimulatory properties, which play a central role in their function as therapeutic agents, have not yet been adequately studied. Here, key aspects of human immune cell response to MTB strain Magnetospirillum magneticum AMB‐1 in physiological environments are characterized. The ability of MTB to maintain magnetic properties, remain viable in whole blood, elicit cytokine production by macrophages, and stimulate uptake of cancer cell material by dendritic cells is examined. This study also investigates the use of MTB–liposome complexes for effective delivery of therapeutic payloads in vitro and explores response to the agent in vivo. Overall, this work establishes the potential of MTB as a versatile, combined delivery platform for immune‐mediated cancer therapy.

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

Cited by 131 publications

References 30 publications

Dose-dependent phorbol 12-myristate-13-acetate-mediated monocyte-to-macrophage differentiation induces unique proteomic signatures in THP-1 cells

Dose-dependent phorbol 12-myristate-13-acetate-mediated monocyte-to-macrophage differentiation induces unique proteomic signatures in THP-1 cells

Effects of nanoparticle‑mediated delivery of pitavastatin on atherosclerotic plaques in ApoE‑knockout mice and THP‑1‑derived macrophages

Engineering Living Immunotherapeutic Agents for Improved Cancer Treatment

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