THP-1 and human peripheral blood mononuclear cell-derived macrophages differ in their capacity to polarize in vitro

Yao

Int Forum Allergy Rhinol

et al. 2018

Background: M2 macrophages are characterized by high interleukin-10 (IL-10) expression and are critical for resolving inflammation. Although increased accumulation of M2 macrophages has been demonstrated in chronic rhinosinusitis with nasal polyps (CRSwNP), particularly the eosinophilic type, their functional relevance in CRSwNP remains poorly understood.Methods: M1 and M2 macrophages and IL-10 expression in sinonasal tissues were detected by doubleimmunofluorescence staining. THP-1 cells, a human monocytic leukemia cell line, were stimulated with various cytokines to study macrophage polarization and IL-10 expression. Polyp size, computed tomography (CT) scans, and symptom severity were scored.Results: Compared with numbers in control tissues, the numbers of total CD68 + macrophages, interferon regulatory factor 5-positive and CD68 + M1 macrophages, and CD163 + CD68 + and CD206 + CD68 + M2 macrophages were increased in both eosinophilic and non-eosinophilic polyps. However, compared with non-eosinophilic polyps, eosinophilic polyps contained fewer M1 macrophages and more M2 macrophages. Consistent with this, the M1/M2 macrophage ratio was increased in non-eosinophilic polyps, whereas it decreased in eosinophilic polyps. Strikingly, the numbers of IL-10 + CD68 + macrophages and the percentage of IL-10 + CD68 + macrophages relative to the total number of macrophages were decreased in eosinophilic polyps, despite the upregulation of M2 macrophages in this type of polyp. The number of IL-10 + CD68 + M2 macrophages correlated negatively with total symptoms scores, polyp sizes, total CT scores, and the total number of inflammatory cells in patients with eosinophilic CRSwNP. Poly I:C downregulated IL-10 expression in M2 macrophages differentiated from THP-1 cells in vitro.Conclusion: Impaired IL-10 production by M2 macrophages may contribute to sustained inflammation in eosinophilic CRSwNP.vv C 2018 ARS-AAOA, LLC.

Section: Methodsmentioning

confidence: 99%

Deficiency in interleukin‐10 production by M2 macrophages in eosinophilic chronic rhinosinusitis with nasal polyps

Yao

Int Forum Allergy Rhinol

et al. 2018

“…The human leukemia monocytic THP‐1 cell line was obtained from the American Type Culture Collection (Rockville, MD) and cultured in RPMI 1640 with 10% fetal bovine serum and 1% penicillin‐streptomycin (Invitrogen) in a humidified incubator containing 5% CO 2 at 37°C . THP‐1 cells were differentiated into macrophages by treatment with 10 ng/mL phorbol‐12‐myristate‐13‐acetate (Sigma‐Aldrich) for 48 hours.…”

Section: Methodsmentioning

confidence: 99%

Hemokinin‐1 stimulates C‐C motif chemokine ligand 24 production in macrophages to enhance eosinophilic inflammation in nasal polyps

Deng

Int Forum Allergy Rhinol

et al. 2019

Background:The mechanisms underlying mucosal eosinophilia in chronic rhinosinusitis with nasal polyps (CRSwNP) remain poorly clarified. The nervous system and neuropeptides play an important role in the regulation of immune response. Herein we explore the expression and function of hemokinin-1 (HK-1), a newly identified tachykinin, along with its receptor neurokinin 1 receptor (NK1R) in CRSwNP. Methods:HK-1, NK1R, and C-C motif chemokine ligand 24 (CCL24) expression in nasal tissues (53 eosinophilic CRSwNP, 32 non-eosinophilic CRSwNP, and 33 controls) was investigated by quantitative reverse transcript polymerase chain reaction (RT-PCR) and immunofluorescence staining. THP-1, a human monocytic leukemia cell line, and eosinophilic polyp tissues were stimulated with HK-1. Cells, tissues, and culture supernatants were subsequently collected for detection of the production of various inflammatory cytokines and chemokines by quantitative RT-PCR and enzyme-linked immunoassay.Results: HK-1 and NK1R mRNA and protein expression were upregulated in eosinophilic and non-eosinophilic nasal polyps compared with control tissues, with eosinophilic polyps demonstrating a higher upregulation compared with that of non-eosinophilic polyps. Eosinophils constituted the major source of HK-1, whereas macrophages were the predominant cell type exhibiting NK1R in eosinophilic polyps. HK-1 induced CCL24 production from macrophages differentiated from THP-1 cells; this was abolished by an NK1R antagonist. HK-1 also induced CCL24 production from ex vivo-cultured eosinophilic nasal polyps. CCL24 was expressed by macrophages in eosinophilic but not noneosinophilic polyps. The expression level of HK-1 correlated with CCL24 expression and tissue eosinophilia in eosinophilic nasal polyps. Conclusion:Eosinophil-derived HK-1 induces CCL24 production from macrophages and therefore exaggerates eosinophilic inflammation in CRSwNP. C 2019 ARS-AAOA, LLC.

J Cellular Molecular Medi

“…Subsequently, PBMCs suspended in serum‐free RPMI 1640 medium (Gibco) supplemented with 1% penicillin/streptomycin were seeded in a six‐well plate for 1 hour in a humidified incubator containing 5% CO 2 at 37°C to allow monocyte adhesion. Nonadherent cells were removed and the adherent monocytes were further incubated in RPMI 1640 medium supplemented with 10% (v/v) heat‐inactivated human serum and 1% penicillin/streptomycin for 7 days and media replacement every 3 days to obtain matured macrophages …”

Section: Methodsmentioning

confidence: 99%

“…Nonadherent cells were removed and the adherent monocytes were further incubated in RPMI 1640 medium supplemented with 10% (v/v) heat-inactivated human serum and 1% penicillin/streptomycin for 7 days and media replacement every 3 days to obtain matured macrophages. 17…”

Section: Peripheral Blood Mononuclear Cells (Pbmcs) Isolation and Mmentioning

confidence: 99%

Helicobacter pylori‐induced exosomal MET educates tumour‐associated macrophages to promote gastric cancer progression

Che

Geng

et al. 2018

Helicobacter pylori (H. pylori) infection triggers chronic inflammation that has been associated with gastric cancer (GC). Exosomes are small extracellular vesicles that have become the key mediators of intercellular communication. In this study, we investigated exosome‐mediated communication between H. pylori‐infected GC cells and macrophages, focusing on the transfer of activated mesenchymal‐epithelial transition factor (MET). We observed a significant decrease in MET protein expression in GC cells after infection with H. pylori, whereas MET mRNA levels remained unchanged. Intriguingly, MET expression, specifically the phosphorylated active form, was increased in exosomes released from H. pylori‐infected GC cells. Confocal microscopy and Western blotting analyses showed that these exosomes containing MET were delivered to and internalized by macrophages. Indeed, in human GC tissues positive for H. pylori, we also observed that activated MET was highly expressed in tumour‐infiltrating macrophages. After internalization, exosomal MET then appeared to educate the macrophages towards a pro‐tumorigenesis phenotype. This included exosomal MET‐mediated stimulation of proinflammatory cytokine secretion IL‐1β, which subsequently promoted tumour growth and progression in vitro and in vivo. Taken together, these data were the first to demonstrate H. pylori infection‐induced upregulation of activated MET in exosomes and the pro‐tumorigenic effect on tumour‐associated macrophages.