Characterization of lipopolysaccharide-stimulated cytokine expression in macrophages and monocytes

Huang, Haiqiu; Fletcher, Arnetta; Niu, Yuge; Wang, Thomas T.Y.; Yu, Liangli

doi:10.1007/s00011-012-0533-8

Cited by 41 publications

(43 citation statements)

References 23 publications

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“…The stability of cytokines in CM has been demonstrated by measuring both cytokine mRNA in the macrophages and cytokine proteins in CM. These studies demonstrated that cellular mRNA was decreased when cytokine protein in CM was maximal (26, 50, 59). For example, in both Wong et al .…”

Section: Introductionmentioning

confidence: 95%

“…M1 macrophages are characterized by high IL-12, IL-6, IL-1, IL-23, TNF-α, nitric oxide production, and increased intracellular pathogen killing (11, 12, 17–19). Other cytokines produced by pro-inflammatory monocytes/macrophages include regulated on activation normal T cell expressed and secreted (RANTES) (20, 21), macrophage inflammatory protein-1β (MIP-1β) (22), macrophage inflammatory protein 2 (MIP-2) (23), tissue inhibitor of metalloproteinase 1 (TIMP-1) (24), interferon-γ-induced protein 10 (IP-10) (17, 25), keratinocyte-derived cytokine (KC-GRO) (23), monocyte chemotactic protein (MCP) MCP-1 (26–29), MCP-3 (27, 29), and MCP-5 (30). Cytokines are generally classified as pro-inflammatory due to their production by immune cells in response to inflammatory stimuli but also by their capability to promote inflammation.…”

Section: Introductionmentioning

confidence: 99%

“…The best example is TNF-α; which is both produced by macrophages in response to LPS and promotes fever (31), vasodilation, leukocyte vascular adhesion, and immune cell recruitment (32). Other inflammatory monocyte/macrophage-related cytokines also have pro-inflammatory functions including IL-12 inducing Th1 cell maturation/activation (33), IL-1 (34) and IL-6 (31) causing fevers, IL-6 stimulating the acute phase response (35), and immune cell recruitment by multiple cytokines: MCP-1 (26–29), MCP-3 (27, 29), MCP-5 (30), IP-10 (36), KC-GRO (17, 37), MIP-1β (17, 38, 39), MIP-2 (23), and RANTES (20, 21). …”

Section: Introductionmentioning

confidence: 99%

“…(59) and Huang et al . (26), TNF-α cellular mRNA and CM protein were measured at the same time points, demonstrating three important issues; 1) TNF-α protein was stable in the CM, 2) TNF-α protein in the CM did not reflect temporal changes in the expression of cellular TNF-α mRNA, 3) sustained elevations of TNF-α in the CM were associated with decreased cellular mRNA possibly due to feedback inhibitory mechanisms (60). Similar observations were also present with IL-6 (26) and IL-12p40 (50).…”

Section: Introductionmentioning

confidence: 99%

“…(26), TNF-α cellular mRNA and CM protein were measured at the same time points, demonstrating three important issues; 1) TNF-α protein was stable in the CM, 2) TNF-α protein in the CM did not reflect temporal changes in the expression of cellular TNF-α mRNA, 3) sustained elevations of TNF-α in the CM were associated with decreased cellular mRNA possibly due to feedback inhibitory mechanisms (60). Similar observations were also present with IL-6 (26) and IL-12p40 (50). Thus, activation of pathways necessary to create the cytokines in the CM likely occurred earlier than the rise in cytokines in the CM, suggesting that signal transduction pathways and release kinetics of cytokines should be studied early in the time course of the experiments after identification of the critical time points for each cytokine.…”

Section: Introductionmentioning

confidence: 99%

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Temporal phenotypic features distinguish polarized macrophagesin vitro

Melton¹,

McManus²,

Gelfond

et al. 2015

Autoimmunity

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Macrophages are important in vascular inflammation and environmental factors influence macrophage plasticity. Macrophage transitions into pro-inflammatory (M1) or anti-inflammatory (M2) states have been defined predominately by measuring cytokines in culture media (CM). However, temporal relationships between cellular and secreted cytokines have not been established. We measured phenotypic markers and cytokines in cellular and CM of murine bone marrow-derived macrophages at multiple time points following stimulation with IFN-γ+LPS (M1), IL-4 (M2a), or IL-10 (M2c). Cytokines/proteins in M1-polarized macrophages exhibited two distinct temporal patterns; an early (0.5–3 hr), transient increase in cellular cytokines (GM-CSF, KC-GRO, MIP-2, IP-10 and MIP-1β) and a delayed (3–6 hrs) response that was more sustained [IL-3, regulated on activation normal T cell expressed and secreted (RANTES), and tissue inhibitor of metalloproteinases 1 (TIMP-1)]. M2a-related cytokine/cell markers (IGF-1, Fizz1, and Ym1) were progressively (3–24 hrs) increased post-stimulation. Additionally, novel patterns were observed. First, and unexpectedly, cellular pro-inflammatory chemokines, MCP-1 and MCP-3 but not MCP-5, were comparably increased in M1 and M2a macrophages. Second, Vegfr1 mRNA was decreased in M1 and increased in M2a macrophages. Finally, VEGF-A was increased in the CM of M1 cultures and strikingly reduced in M2a coinciding with increased Vegfr1 expression, suggesting decreased VEGF-A in M2a CM was secondary to increased soluble VEGFR1. In conclusion, macrophage cytokine production and marker expression were temporally regulated and relative levels compared across polarizing conditions were highly dependent upon the timing and location (cellular vs. CM) of the sample collection. For most cytokines, cellular production preceded increases in the CM suggesting that cellular regulatory pathways should be studied within 6 hours of stimulation. The divergent polarization-dependent expression of Vegfr1 may be essential to controlling VEGF potentially regulating angiogenesis and inflammatory cell infiltration in the vascular niche. The current study expands the repertoire of cytokines produced by polarized macrophages and provides insights into the dynamic regulation of macrophage polarization and resulting cytokines, proteins, and gene expression that influence vascular inflammation.

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Section: Introductionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Temporal phenotypic features distinguish polarized macrophagesin vitro

Melton¹,

McManus²,

Gelfond

et al. 2015

Autoimmunity

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Direct electrical stimulation enhances osteogenesis by inducing Bmp2 and Spp1 expressions from macrophages and preosteoblasts

Srirussamee

Mobini

Cassidy

et al. 2019

Biotech & Bioengineering

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The capability of electrical stimulation (ES) in promoting bone regeneration has already been addressed in clinical studies. However, its mechanism is still being investigated and discussed. This study aims to investigate the responses of macrophages (J774A.1) and preosteoblasts (MC3T3‐E1) to ES and the faradic by‐products from ES. It is found that pH of the culture media was not significantly changed, whereas the average hydrogen peroxide concentration was increased by 3.6 and 5.4 µM after 1 and 2 hr of ES, respectively. The upregulation of Bmp2 and Spp1 messenger RNAs was observed after 3 days of stimulation, which is consistent among two cell types. It is also found that Spp1 expression of macrophages was partially enhanced by faradic by‐products. Osteogenic differentiation of preosteoblasts was not observed during the early stage of ES as the level of Runx2 expression remains unchanged. However, cell proliferation was impaired by the excessive current density from the electrodes, and also faradic by‐products in the case of macrophages. This study shows that macrophages could respond to ES and potentially contribute to the bone formation alongside preosteoblasts. The upregulation of Bmp2 and Spp1 expressions induced by ES could be one of the mechanisms behind the electrically stimulated osteogenesis.

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Transcriptional and translational‐uncoupling in regulation of the CXCL12 and its receptors CXCR4, 7 in THP‐1 monocytes and macrophages

Yu¹,

Yu²,

Pham

et al. 2017

Immunity Inflam & Disease

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IntroductionThe chemokine CXCL12 and its receptors CXCR4 and 7 play crucial roles in the immune system. In the present study, regulation of this pathway was further examined using the in‐vitro model of undifferentiated human THP‐1 monocytes (u‐THP‐1) and phorbol 12‐myristate 13‐acetate (PMA)‐differentiated THP‐1 macrophages (d‐THP‐1), to assess the effects of differentiation and the TLR4 ligand lipopolysaccharide (LPS) on the pathway.Methods/ResultsDifferentiation did not affect the CXCR4, 7 mRNA levels. Interestingly, the CXCL12 and CXCR7 proteins but not CXCR4 were found to be up‐regulated during differentiation. LPS, through CD14‐dependent pathway, induced CXCL12 and CXCR4, 7 mRNA levels to a greater magnitude in d‐ than u‐THP‐1. The induction effect on CXCL12 stimulated by LPS was confirmed using ELISA. Increased migration of u‐THP‐1 was observed using conditioned medium from LPS‐treated d‐THP‐1. Additionally, d‐THP‐1, although expressed higher CXCR7 protein levels, failed to migrate toward CXCL12. In contrast, LPS did not affect CXCR4, 7 protein levels.ConclusionHence, this study indicated that CXCL12, CXCR4, and CXCR7 were differentially expressed and regulated in u‐THP‐1 and d‐THP‐1 cells in response to external stimuli. Importantly, we reported here a novel observation that uncoupling exists between transcriptional and translational regulation of CXCR4, 7 expressions by differentiation and TLR stimuli.

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Characterization of lipopolysaccharide-stimulated cytokine expression in macrophages and monocytes

Abstract: In summary, this study indicated that temporal and dose-dependent responses to LPS need to be controlled for and that extrapolation of data on mechanisms may differ between cell lines of different origin.

Cited by 41 publications

References 23 publications

Temporal phenotypic features distinguish polarized macrophagesin vitro

Temporal phenotypic features distinguish polarized macrophagesin vitro

Direct electrical stimulation enhances osteogenesis by inducing Bmp2 and Spp1 expressions from macrophages and preosteoblasts

Transcriptional and translational‐uncoupling in regulation of the CXCL12 and its receptors CXCR4, 7 in THP‐1 monocytes and macrophages

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