Recombinant Human Proteoglycan 4 Regulates Phagocytic Activation of Monocytes and Reduces IL-1β Secretion by Urate Crystal Stimulated Gout PBMCs

Elsayed, Sandy; Jay, Gregory D.; Cabezas, Ralph; Qadri, Marwa; Schmidt, Tannin A.; Elsaid, Khaled A.

doi:10.3389/fimmu.2021.771677

Cited by 11 publications

(14 citation statements)

References 62 publications

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“…THP-1 designates a spontaneously immortalized monocyte-like cell line, which has been widely used in studies of gout. Pam3CSK4 (50), PMA (51), LPS (52), and IL-6 (53) were all used for priming the THP-1 cells. Bone marrow derived monocytes (BMDM) are murine primary cells, for which priming is necessary as well.…”

Section: Discussionmentioning

confidence: 99%

Recent Insights Into the Role of Macrophages in Acute Gout

et al. 2022

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Gout is a common type of inflammatory arthritis characterized by the presence of monosodium urate crystals (MSU) in the joints. Macrophages are believed to be involved in gout flares. It has long been recognized that resident macrophage and monocyte derived macrophages are distinct subsets and there have been attempts to investigate their roles in acute gout, respectively. Previous studies revealed that resident macrophages initiate and drive the inflammation, while monocyte derived macrophages differentiated into M1-like macrophages in response to MSU crystals. With the advancement of technologies, subpopulations of synovial resident macrophages have been defined with the characteristics more accurately described. Resident macrophages in the synovial lining layer showed an anti-inflammatory effect in rheumatoid arthritis, but specific Trpv4 depletion of them reduced MSU crystals induced murine arthritis. CD14+ monocytes in the synovial fluid from patients with gout exhibit phenotypes of anti-inflammatory as well as pro-inflammatory characteristics. Here, we review the main aspects of macrophages in the initiation and resolution of acute gout and try to clarify the specific role of each subpopulation. Building a reliable diagram of the effect of monocytes and macrophages during MSU crystals induced arthritis will bring us closer to targeting macrophages for improving the management of gout.

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Section: Discussionmentioning

confidence: 99%

Recent Insights Into the Role of Macrophages in Acute Gout

et al. 2022

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“…Moreover, a variety of studies reported that the maturation and release of IL-1β were related to the activation of the NLRP3 inflammasome, and activation of NLRP3 was an important pathway for MSU to contribute to inflammatory responses. Thus, we speculated whether the anti-inflammatory mechanism of neoastilbin was connected with the NLRP3 inflammasome [ 39 , 40 ]. In vitro and in vivo experiments showed that the expression of NLRP3, Caspase-1 and ASC proteins increased significantly after MSU stimulation, indicating that the NLRP3 inflammasome might be activated.…”

Section: Discussionmentioning

confidence: 99%

The Protective Effects of Neoastilbin on Monosodium Urate Stimulated THP-1-Derived Macrophages and Gouty Arthritis in Mice through NF-κB and NLRP3 Inflammasome Pathways

Zhang

et al. 2022

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Gouty arthritis (GA) is a frequent inflammatory disease characterized by pain, swelling, and stiffness of joints. Neoastilbin is a flavonoid isolated from the rhizome of Smilax glabra, which possesses various anti-inflammatory effects. However, the mechanism of neoastilbin in treating GA has not yet been clarified. Thus, this study was to investigate the protective effects of neoastilbin in both monosodium urate (MSU) stimulated THP-1-derived macrophages and the animal model of GA by injecting MSU into the ankle joints of mice. The levels of key inflammatory cytokines in MSU stimulated THP-1-derived macrophages were detected by enzyme-linked immunosorbent assay (ELISA) kits. Protein expressions of nuclear factor kappa B (NF-κB) and NOD-like receptor protein 3 (NLRP3) inflammasome pathways were further detected by Western blotting. In addition, swelling degree of ankle joints, the levels of inflammatory factors, infiltration of inflammatory cells and the expressions of related proteins were determined. Swelling degree and histopathological injury in ankle joints of MSU-injected mice were significantly decreased after being treated with neoastilbin. Moreover, neoastilbin significantly diminished the secretion of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α), suppressing the activation of NF-κB and NLRP3 inflammasome pathways in both MSU stimulated THP-1-derived macrophages and the mouse model of GA. In summary, neoastilbin could alleviate GA by inhibiting the NF-κB and NLRP3 inflammasome pathways, which provided some evidence for neoastilbin as a promising therapeutic agent for GA treatment.

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“…We employed the murine peritoneal model of acute gout as previously described ( Elsayed et al, 2021 ). Our experiments were approved by the IACUC committee at Chapman University.…”

Section: Methodsmentioning

confidence: 99%

“…Pyrogen-free MSU crystals (2 mg in 200 μL), vehicle (200 μL) or fingolimod (5 mg/kg; 200 μL) were administered intraperitoneally and peritoneal lavaging was performed at 4 h. Lavaging was performed by injecting 3 ml of cold DPBS into the peritoneal cavity followed by shaking for 30–60 s and lavage aspiration. Lavage fluids were centrifuged at 3,000 rpm for 5 min at 4°C, and cell pellets were resuspended in PBS and immunophenotyping of CMs, NCMs and neutrophils was conducted as previously described ( Elsayed et al, 2021 ). CMs were identified as CD11b+ Ly6C hi CCR2+, while NCMs were identified as CD11b+ Ly6C lo CD43 hi CX3CR1+ and neutrophils were identified as Ly6G + Ly6B.2+ ( Elsayed et al, 2021 ).…”

Section: Methodsmentioning

confidence: 99%

“…CD11b+ cells were identified and two populations: Ly6C hi and Ly6C lo cells were gated. We subsequently gated for CCR2 positivity in the Ly6C hi population Frontiers in Pharmacology frontiersin.org to identify CMs and gated for CD43 and CX3CR1 in the Ly6C lo population to identify NCMs (Elsayed et al, 2021). For neutrophils, singlet viable cells were gated for Ly6G and Ly6B.2 markers.…”

Section: M1 and M2 Markers' Expressions And Cytokine Array Assays In ...mentioning

confidence: 99%

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Protein phosphatase 2A regulates xanthine oxidase-derived ROS production in macrophages and influx of inflammatory monocytes in a murine gout model

Elsayed

Elsaid²

2022

Front. Pharmacol.

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Background: Gout is a common arthritis, due to deposition of monosodium urate (MSU) crystals which results in IL-1β secretion by tissue-resident macrophages. Xanthine oxidase (XO) catalyzes uric acid (UA) production and in the process, reactive oxygen species (ROS) are generated which contributes to NLRP3 inflammasome activation. Protein phosphatase 2A (PP2A) may be involved in regulating inflammatory pathways in macrophages. The objective of this study was to investigate whether PP2A regulates gout inflammation, mediated by XO activity modulation. We studied UA and ROS generations in MSU stimulated murine bone marrow derived macrophages (BMDMs) in response to fingolimod phosphate, a PP2A activator, and compared its anti-inflammatory efficacy to that of an XO inhibitor, febuxostat.Methods: BMDMs were stimulated with MSU, GM-CSF/IL-1β or nigericin ± fingolimod (2.5 μM) or febuxostat (200 μM) and UA levels, ROS, XO, and PP2A activities, Xdh (XO) expression and secreted IL-1β levels were determined. PP2A activity and IL-1β in MSU stimulated BMDMs ± N-acetylcysteine (NAC) (10 μM) ± okadaic acid (a PP2A inhibitor) were also determined. M1 polarization of BMDMs in response to MSU ± fingolimod treatment was assessed by a combination of iNOS expression and multiplex cytokine assay. The in vivo efficacy of fingolimod was assessed in a murine peritoneal model of acute gout where peritoneal lavages were studied for pro-inflammatory classical monocytes (CMs), anti-inflammatory nonclassical monocytes (NCMs) and neutrophils by flow cytometry and IL-1β by ELISA.Results: Fingolimod reduced intracellular and secreted UA levels (p < 0.05), Xdh expression (p < 0.001), XO activity (p < 0.001), ROS generation (p < 0.0001) and IL-1β secretion (p < 0.0001), whereas febuxostat enhanced PP2A activity (p < 0.05). NAC treatment enhanced PP2A activity and reduced XO activity and PP2A restoration mediated NAC’s efficacy as co-treatment with okadaic acid increased IL-1β secretion (p < 0.05). Nigericin activated caspase-1 and reduced PP2A activity (p < 0.001) and fingolimod reduced caspase-1 activity in BMDMs (p < 0.001). Fingolimod reduced iNOS expression (p < 0.0001) and secretion of IL-6 and TNF-α (p < 0.05). Fingolimod reduced CMs (p < 0.0001), neutrophil (p < 0.001) and IL-1β (p < 0.05) lavage levels while increasing NCMs (p < 0.001).Conclusion: Macrophage PP2A is inactivated in acute gout by ROS and a PP2A activator exhibited a broad anti-inflammatory effect in acute gout in vitro and in vivo.

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Recombinant Human Proteoglycan 4 Regulates Phagocytic Activation of Monocytes and Reduces IL-1β Secretion by Urate Crystal Stimulated Gout PBMCs

Cited by 11 publications

References 62 publications

Recent Insights Into the Role of Macrophages in Acute Gout

Recent Insights Into the Role of Macrophages in Acute Gout

The Protective Effects of Neoastilbin on Monosodium Urate Stimulated THP-1-Derived Macrophages and Gouty Arthritis in Mice through NF-κB and NLRP3 Inflammasome Pathways

Protein phosphatase 2A regulates xanthine oxidase-derived ROS production in macrophages and influx of inflammatory monocytes in a murine gout model

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