Assessment of Phagocytic Activity of Cultured Macrophages Using Fluorescence Microscopy and Flow Cytometry

Sharma, Lokesh; Wu, Wenjun; Dholakiya, Sanjay L.; Gorasiya, Samir; Wu, Jiao; Sitapara, Ravikumar; Patel, Vivek; Wang, Mao; Zur, Michelle; Reddy, Shloka; Siegelaub, Nathan; Bamba, Katrina; Barile, Frank A.; Mantell, Lin L.

doi:10.1007/978-1-4939-0928-5_12

Cited by 28 publications

(18 citation statements)

References 34 publications

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“…TGF-β is a potent suppressor of Th1 and Th2 effector cell differentiation, a regulator of Foxp3 + regulatory T cells, and is a critical cytokine for the induction of Th9 and Th17 cells [ 66 – 68 ]. TGF-β also has regulatory effects on CD8 + cells [ 69 ] and promotes phagocytosis [ 70 ]. Thus, we propose that increased activation of the TGF-β pathway in elderly septic patients may induce a prolonged downregulation of adaptive immunity and an increased Th17 response.…”

Section: Discussionmentioning

confidence: 99%

Septic Shock in Advanced Age: Transcriptome Analysis Reveals Altered Molecular Signatures in Neutrophil Granulocytes

et al. 2015

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Sepsis is one of the highest causes of mortality in hospitalized people and a common complication in both surgical and clinical patients admitted to hospital for non-infectious reasons. Sepsis is especially common in older people and its incidence is likely to increase substantially as a population ages. Despite its increased prevalence and mortality in older people, immune responses in the elderly during septic shock appear similar to that in younger patients. The purpose of this study was to conduct a genome-wide gene expression analysis of circulating neutrophils from old and young septic patients to better understand how aged individuals respond to severe infectious insult. We detected several genes whose expression could be used to differentiate immune responses of the elderly from those of young people, including genes related to oxidative phosphorylation, mitochondrial dysfunction and TGF-β signaling, among others. Our results identify major molecular pathways that are particularly affected in the elderly during sepsis, which might have a pivotal role in worsening clinical outcomes compared with young people with sepsis.

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Section: Discussionmentioning

confidence: 99%

Septic Shock in Advanced Age: Transcriptome Analysis Reveals Altered Molecular Signatures in Neutrophil Granulocytes

et al. 2015

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“…2A-C, Supplementary Fig. S2A-C), possibly because of their phagocytic activity 36 in comparison to the other cell types investigated. As shown from the 3D-rendered images, the cellular uptake of single short tubes or small agglomerates occurs in all three cell types, while large agglomerates are mainly attached on the cells surface ( Fig.…”

Section: Mwcnt-lung Cell Interactionmentioning

confidence: 95%

Profibrotic Activity of Multiwalled Carbon Nanotubes Upon Prolonged Exposures in Different Human Lung Cell Types

Chortarea

Zerimariam

Barošová

et al. 2019

Applied In Vitro Toxicology

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Introduction: The expanding use of multiwalled carbon nanotubes (MWCNTs) in various consumer and industrial products has raised concerns regarding the potential health risks following the inhalation of this nanomaterial. Recent studies indicate that MWCNT exposure can cause deleterious lung effects. Pulmonary fibrosis is one of the diseases associated with the inhalation of classical fiber-shaped materials (e.g., asbestos), and recently MWCNTs. Materials and Methods: Herein, the presented work investigated early signs of fibrosis in key human lung cell types implicated directly (fibroblasts) or indirectly (alveolar epithelial cells and macrophages) in the pathogenesis of lung fibrosis, upon acute (24 hours) and prolonged (96 hours) exposure to different concentrations (5-20 lg/mL) of long, rigid Mitsui-7 MWCNTs. These results were compared to those inflicted upon the same cell types following exposure to tangled nanofibers, that is, Nanocyl-7000, and a positive particle control, crystalline quartz (i.e., Min-U-Sil). Results: Mitsui-7 interacted with all cell types tested, in particular, single nanotubes were readily internalized by each cell type. Exposure to Mitsui-7 resulted in a proinflammatory response in macrophages at 24 hours (interleukin-1b release). However, only prolonged Mitsui-7 exposure induced a significant release of profibrotic mediators (transforming growth factor-b, platelet derived-growth factor, and osteopontin) in all tested cell cultures and concentrations, highlighting the importance of prolonged exposure studies in nanosafety assessment. This elevated release of profibrotic markers was not accompanied by an increased collagen production or cell proliferation in fibroblasts in the tested time frame. Conclusion: Overall, prolonged exposure to human-relevant CNT concentrations appears to be a highly suitable approach to predict profibrotic response in vitro.

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“…The phagocytosis assay was determined as previously described (Sharma, Wu, & Dholakiya, ). Murine macrophages were incubated with Saccharomyces cerevisiae yeasts (5 × 10 6 yeast/well) and 100 μg/mL propidium iodide for 24 h. Subsequently, the phagocytosed yeast was estimated by fluorescence in a flow cytometer (Cell Lab Quanta SC, Beckman Coulter, Fullteron, CA).…”

Section: Methodsmentioning

confidence: 99%

Anti‐inflammatory and antinociceptive effects of tilifodiolide, isolated from Salvia tiliifolia Vahl (Lamiaceae)

González‐Chávez

Alonso-Castro

Zapata-Morales

et al. 2018

Drug Development Research

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Salvia tiliifolia Vahl (Lamiaceae) is used for the empirical treatment of pain and inflammation. The diterpenoid tilifodiolide (TFD) was isolated from Salvia tiliifolia. The in vitro anti-inflammatory effects of TFD (0.1-200 µM) were assessed using murine macrophages stimulated with LPS and estimating the levels of pro-inflammatory mediators for 48 h. The in vivo anti-inflammatory activity of TFD was assessed using the carrageenan-induced paw edema test for 6 h. The antinociceptive effects of TFD were evaluated using the formalin test and the acetic acid induced-writhing test. The effects of TFD on locomotor activity were assessed using the open field test and the rotarod test. TFD inhibited the production of TNF-α (IC = 5.66 µM) and IL-6 (IC = 1.21 µM) in macrophages. TFD (200 mg/kg) showed anti-inflammatory effects with similar activity compared to 10 mg/kg indomethacin. The administration of TFD induced antinociception in the phase 1 (ED = 48.2 mg/kg) and the phase 2 (ED = 28.9 mg/kg) of the formalin test. In the acetic acid assay, TFD showed antinociceptive effects (ED = 32.3 mg/kg) with similar potency compared to naproxen (ED = 36.2 mg/kg). In the presence of different inhibitors in the acetic acid assay, only the co-administration of TFD and naloxone reverted the antinociceptive activity shown by TFD alone. TFD did not affect locomotor activity in mice. TFD exerts in vitro and in vivo anti-inflammatory activity and in vivo antinociceptive effects.

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Assessment of Phagocytic Activity of Cultured Macrophages Using Fluorescence Microscopy and Flow Cytometry

Cited by 28 publications

References 34 publications

Septic Shock in Advanced Age: Transcriptome Analysis Reveals Altered Molecular Signatures in Neutrophil Granulocytes

Septic Shock in Advanced Age: Transcriptome Analysis Reveals Altered Molecular Signatures in Neutrophil Granulocytes

Profibrotic Activity of Multiwalled Carbon Nanotubes Upon Prolonged Exposures in Different Human Lung Cell Types

Anti‐inflammatory and antinociceptive effects of tilifodiolide, isolated from Salvia tiliifolia Vahl (Lamiaceae)

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