scholarly journals Continuous Exposure to Low Doses of Ultrafine Black Carbon Reduces the Vitality of Immortalized Lung-Derived Cells and Activates Senescence

2020 ◽  
Vol 2020 ◽  
pp. 1-13
Author(s):  
M. Esther Salinas ◽  
Denisse A. Gutiérrez ◽  
Armando Varela-Ramírez ◽  
Kristine M. Garza
Keyword(s):  
Black Carbon ◽  
Culture Media ◽  
Human Peripheral Blood ◽  
Cell Types ◽  
Defense Responses ◽  
Continuous Exposure ◽  
Dependent Manner ◽  
Blood Monocytes ◽  
Cellular Modifications ◽  
The Impact

Combustion-derived nanomaterials are noxious ultrafine (<100 nm) aerosol by-products of human activity. They pose threats to pulmonary health due to their small size, allowing them to penetrate alveoli causing detrimental responses downstream. Information regarding the cellular activity that connects nanocarbon particle exposure to poor pulmonary health remains lacking. We hypothesized that low-dose and long-term administrations of carbonaceous nanoparticles contribute to lung irritation by adversely affecting respiratory cells that function as the first line of defense. Responses to ultrafine black carbon (UBC), a key component of airborne pollutants, by human lung A549, murine lung LA4 epithelial cells, human peripheral-blood monocytes THP1, and murine macrophages RAW264.7 were investigated. The cells were first plated on day zero and were fed fresh UBC suspended in culture media on days one, four, and seven. The exposure regimen included three different concentrations of UBC. On day ten, all cells were harvested, washed, and assayed. The impact on cellular viability revealed that UBC was only moderately cytotoxic, while metabolic activity was significantly diminished in a dose-dependent manner. Additionally, beta-galactosidase proportionally increased with UBC concentration compared to untreated cells, indicating that cellular senescence was promoted across all cell types. The implemented regimen caused minimal toxicity yet demonstrated different cellular modifications across the cell lines of both species, inducing changes to enzyme vitality and cellular fitness. The data suggested that compounding nanosized black carbon exposure could negatively impair overall pulmonary health by distinctively modifying intracellular behavior.

Toward functional proteomics of alveolar macrophages

2005 ◽  
Vol 288 (4) ◽  
pp. L585-L595 ◽  
Author(s):  
Haifeng M. Wu ◽  
Ming Jin ◽  
Clay B. Marsh
Keyword(s):  
Alveolar Macrophages ◽  
Large Scale ◽  
Cell Types ◽  
Mononuclear Phagocyte ◽  
Detection Sensitivity ◽  
Current Status ◽  
Blood Monocytes ◽  
Protein Patterns ◽  
Additional Information ◽  
The Impact

Alveolar macrophages (AM) belong to a phenotype of macrophages with distinct biological functions and important pathophysiological roles in lung health and disease. The molecular details determining AM differentiation from blood monocytes and AM roles in lung homeostasis are largely unknown. With the use of different technological platforms, advances in the field of proteomics have made it possible to search for differences in protein expression between AM and their precursor monocytes. Proteome features of each cell type provide new clues into understanding mononuclear phagocyte biology. In-depth analyses using subproteomics and subcellular proteomics offer additional information by providing greater protein resolution and detection sensitivity. With the use of proteomic techniques, large-scale mapping of phosphorylation differences between the cell types have become possible. Furthermore, two-dimensional gel proteomics can detect germline protein variants and evaluate the impact of protein polymorphisms on an individual's susceptibility to disease. Finally, surface-enhanced laser desorption and ionization (SELDI) time-of-flight mass spectrometry offers an alternative method to recognizing differences in protein patterns between AM and monocytes or between AM under different pathological conditions. This review details the current status of this field and outlines future directions in functional proteomic analyses of AM and monocytes. Furthermore, this review presents viewpoints of integrating proteomics with translational topics in lung diseases to define the mechanisms of disease and to uncover new diagnostic and therapeutic targets.

scholarly journals Effect of Clinoptilolite and Sepiolite Nanoclays on Human and Parasitic Highly Phagocytic Cells

2015 ◽  
Vol 2015 ◽  
pp. 1-12 ◽  
Author(s):  
Yanis Toledano-Magaña ◽  
Leticia Flores-Santos ◽  
Georgina Montes de Oca ◽  
Alfonso González-Montiel ◽  
Juan-Pedro Laclette ◽  
...  
Keyword(s):  
Cell Line ◽  
Human Peripheral Blood ◽  
Dependent Manner ◽  
Blood Monocytes ◽  
Phagocytic Cells ◽  
Need To Evaluate ◽  
Potential Applications ◽  
Low Cytotoxicity ◽  
Dose Dependent

Nanoclays have potential applications in biomedicine raising the need to evaluate their toxicity inin vitromodels as a first approach to its biocompatibility. In this study,in vitrotoxicity of clinoptilolite and sepiolite nanoclays (NC) was analyzed in highly phagocytic cultures of amoebas and human and mice macrophages. While amebic viability was significantly affected only by sepiolite NC at concentrations higher than 0.1 mg/mL, the effect on macrophage cultures was dependent on the origin of the cells. Macrophages derived from human peripheral blood monocytes were less affected in viability (25% decrease at 48 h), followed by the RAW 264.7 cell line (40%), and finally, macrophages derived from mice bone marrow monocytes (98%). Moreover, the cell line and mice macrophages die mainly by necrosis, whereas human macrophages exhibit increased apoptosis. Cytokine expression analysis in media of sepiolite NC treated cultures showed a proinflammatory profile (INFγ, IL-1α, IL-8, and IL-6), in contrast with clinoptilolite NC that induced lees cytokines with concomitant production of IL-10. The results show that sepiolite NC is more toxic to amoebas and macrophages than clinoptilolite NC, mostly in a time and dose-dependent manner. However, the effect of sepiolite NC was comparable with talc powder suggesting that both NC have low cytotoxicityin vitro.

scholarly journals Leukotriene B4 stimulates c-fos and c-jun gene transcription and AP-1 binding activity in human monocytes

1992 ◽  
Vol 282 (3) ◽  
pp. 625-629 ◽  
Author(s):  
J Staňková ◽  
M Rola-Pleszczynski
Keyword(s):  
Human Peripheral Blood ◽  
Leukotriene B4 ◽  
Binding Activity ◽  
Dependent Manner ◽  
Blood Monocytes ◽  
Concentration Dependent Manner ◽  
Nuclear Factors ◽  
Oncogene Products ◽  
Kinetics Of

We have examined the effect of leukotriene B4 (LTB4), a potent lipid proinflammatory mediator, on the expression of the proto-oncogenes c-jun and c-fos. In addition, we looked at the modulation of nuclear factors binding specifically to the AP-1 element after LTB4 stimulation. LTB4 increased the expression of the c-fos gene in a time- and concentration-dependent manner. The c-jun mRNA, which is constitutively expressed in human peripheral-blood monocytes at relatively high levels, was also slightly augmented by LTB4, although to a much lower extent than c-fos. The kinetics of expression of the two genes were also slightly different, with c-fos mRNA reaching a peak at 15 min after stimulation and c-jun at 30 min. Both messages rapidly declined thereafter. Stability of the c-fos and c-jun mRNA was not affected by LTB4, as assessed after actinomycin D treatment. Nuclear transcription studies in vitro showed that LTB4 increased the transcription of the c-fos gene 7-fold and the c-jun gene 1.4-fold. Resting monocytes contained nuclear factors binding to the AP-1 element, but stimulation of monocytes with LTB4 induced greater AP-1-binding activity of nuclear proteins. These results indicate that LTB4 may regulate the production of different cytokines by modulating the yield and/or the function of transcription factors such as AP-1-binding proto-oncogene products.

scholarly journals Interaction of Graphene Oxide Modified with Linear and Branched PEG with Monocytes Isolated from Human Blood

Nanomaterials ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 126
Author(s):  
Pavel Khramtsov ◽  
Maria Bochkova ◽  
Valeria Timganova ◽  
Anton Nechaev ◽  
Sofya Uzhviyuk ◽  
...  
Keyword(s):  
Graphene Oxide ◽  
Immune Cells ◽  
Human Peripheral Blood ◽  
Oxide Surface ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Composition And Structure ◽  
Luminescent Signal ◽  
Ros Formation ◽  
The Impact

Multiple graphene-based therapeutics have recently been developed, however potential risks related to the interaction between nanomaterials and immune cells are still poorly understood. Therefore, studying the impact of graphene oxide on various populations of immune cells is of importance. In this work, we aimed to investigate the effects of PEGylated graphene oxide on monocytes isolated from human peripheral blood. Graphene oxide nanoparticles with lateral sizes of 100–200 nm and 1–5 μm were modified with linear and branched PEG (GO-PEG). Size, elemental composition, and structure of the resulting nanoparticles were characterized. We confirmed that PEG was successfully attached to the graphene oxide surface. The influence of GO-PEG on the production of reactive oxygen species (ROS), cytokines, phagocytosis, and viability of monocytes was studied. Uptake of GO-PEG by monocytes depends on PEG structure (linear or branched). Branched PEG decreased the number of GO-PEG nanoparticles per monocyte. The viability of monocytes was not altered by co-cultivation with GO-PEG. GO-PEG decreased the phagocytosis of Escherichia coli in a concentration-dependent manner. ROS formation by monocytes was determined by measuring luminol-, lucigenin-, and dichlorodihydrofluorescein-dependent luminescence. GO-PEG decreased luminescent signal probably due to inactivation of ROS, such as hydroxyl and superoxide radicals. Some types of GO-PEG stimulated secretion of IL-10 by monocytes, but this effect did not correlate with their size or PEG structure.

scholarly journals Differentiation stimuli induce receptors for plasma fibronectin on the human myelomonocytic cell line HL-60

Blood ◽  
1984 ◽  
Vol 64 (4) ◽  
pp. 858-866
Author(s):  
CG Pommier ◽  
J O'Shea ◽  
T Chused ◽  
T Takahashi ◽  
M Ochoa ◽  
...  
Keyword(s):  
Cell Line ◽  
Human Peripheral Blood ◽  
Cell Types ◽  
Plasma Fibronectin ◽  
Blood Monocytes ◽  
Before And After ◽  
Differentiating Agents ◽  
Membrane Antigens ◽  
Human Polymorphonuclear Leukocytes

Plasma fibronectin (Fn) induces phagocytosis of C3b-opsonized sheep erythrocytes (EC3b) by human peripheral blood monocytes. However, Fn does not induce erythrophagocytosis of EC3b by human polymorphonuclear leukocytes (PMN), unless the PMN have been exposed to C5a or N-formyl- methionyl-leucyl-phenylalanine. Because of this difference, it is of great interest to examine Fn binding to cells that possess the capacity to differentiate into either granulocytes or monocytes. Hence, we have examined the consequences of Fn binding to the human myelomonocytic cell line, HL-60, both before and after in vitro differentiation of the HL-60, along a monocytoid or a granulocytoid pathway. Fn receptors were not found on undifferentiated HL-60, but several differentiating agents promoted the HL-60 binding of Fn-coated microspheres (Fn-ms). The peak of Fn-ms binding occurred four to five days after the induction of differentiation with dimethylsulfoxide (DMSO), and two days after induction by PMA. In addition, cells that differentiated along either the monocytoid or the granulocytoid pathway showed a marked increase in the phagocytosis of both IgG-coated erythrocytes (EA) and EC3b when they were exposed to Fn. Comparison of the effects of anti-Fn monoclonals on the binding of Fn-ms to the monocytes, PMN, and HL-60 showed that the same monoclonals block Fn-ms-binding and Fn-induced EC3b phagocytosis by all three cell types. Two monoclonal antibodies, M1/70 and A6F10, directed against membrane antigens on PMN and monocytes, inhibited Fn-ms binding. Both also blocked Fn-induced EC3b ingestion by these cells. However, neither antibody blocked Fn-ms binding or EC3b ingestion by differentiated HL-60. We conclude that differentiated HL-60 cells express functionally active Fn receptors, similar to monocytes and activated PMN, which, nonetheless, differ from normal cells in their association with the antigens recognized by M1/70 and A6F10.

Glutathione peroxidase-1 but not -4 is involved in the regulation of cellular 5-lipoxygenase activity in monocytic cells

2000 ◽  
Vol 349 (2) ◽  
pp. 455-461 ◽  
Author(s):  
Dietlind STRAIF ◽  
Oliver WERZ ◽  
Roland KELLNER ◽  
Ute BAHR ◽  
Dieter STEINHILBER
Keyword(s):  
Glutathione Peroxidase ◽  
Protein Interactions ◽  
Human Peripheral Blood ◽  
Cell Types ◽  
Blood Monocytes ◽  
Phospholipid Hydroperoxide Glutathione Peroxidase ◽  
Strong Inhibitor ◽  
Monocytic Cell ◽  
Monocytic Cell Line ◽  
Glutathione Peroxidase 1

In contrast to neutrophils or B-lymphocytes, cells of the monocytic lineage like rat macrophages, human peripheral blood monocytes and Mono Mac 6 cells contain a strong inhibitor of 5-lipoxygenase (5-LO) activity, which scavenges hydroperoxides and inhibits 5-LO activity in broken-cell preparations in the absence of exogenously added thiols. Chromatographic purification of the inhibitor from the human monocytic cell line Mono Mac 6 and amino acid sequence analysis revealed that the inhibitory factor is glutathione peroxidase-1 (GPx-1). In contrast to the peroxidase activity of GPx-1, 5-LO inhibition by GPx-1 was supported by β-mercaptoethanol and there was no absolute requirement for millimolar concentrations of glutathione or dithiothreitol. These cofactor characteristics suggest that both activities address distinct catalytic properties of GPx-1. 5-LO inhibition by GPx-1 was not due to direct GPx-5-LO protein-protein interactions, since GPx-1 did not bind to immobilized 5-LO. Interestingly, 5-LO derived from granulocytes was significantly more resistant against GPx-1 inhibition than B-lymphocytic 5-LO, which correlates with the respective cellular 5-LO activities. In summary, the data suggest that, in addition to previously reported phospholipid hydroperoxide glutathione peroxidase (GPx-4), GPx-1 is an efficient inhibitor of 5-LO even at low thiol concentrations, and is involved in the regulation of cellular 5-LO activity in various cell types.

scholarly journals Autoregulatory Effect of Interleukin-10 on Proinflammatory Cytokine Production by Porphyromonas gingivalis Lipopolysaccharide-Tolerant Human Monocytes

1999 ◽  
Vol 67 (5) ◽  
pp. 2153-2159 ◽  
Author(s):  
Hidetoshi Shimauchi ◽  
Tomohiko Ogawa ◽  
Kozo Okuda ◽  
Yutaka Kusumoto ◽  
Hirohi Okada
Keyword(s):  
Porphyromonas Gingivalis ◽  
Human Peripheral Blood ◽  
Interleukin 10 ◽  
High Dose ◽  
Dependent Manner ◽  
Blood Monocytes ◽  
High Concentration ◽  
Down Regulation ◽  
Enhanced Production ◽  
Porphyromonas Gingivalis Lipopolysaccharide

ABSTRACT Pretreatment of human peripheral blood monocytes with a very low concentration (0.1 ng/ml) of Porphyromonas gingivalislipopolysaccharides (LPS) resulted in a significant decrease of interleukin-6 (IL-6) production, but not IL-8 production, by restimulation of a high concentration (1 μg/ml) of the same LPS. In contrast, the same pretreatment with Escherichia coli LPS resulted in the enhanced production of both IL-6 and IL-8 after restimulation. The selective induction by P. gingivalis LPS tolerance of IL-6 production developed in a time-dependent manner during the primary culture. P. gingivalis LPS-pretreated cells were also refractory to a high-dose E. coli LPS restimulation in terms of IL-6 production. The expression of IL-6 mRNA decreased 10 h after restimulation of P. gingivalisLPS-pretreated monocytes. Furthermore, an up-regulation of anti-inflammatory cytokine IL-10 upon a second high-dose LPS rechallenge occurred at the same time point in the pretreated cells. We studied the role of IL-10 in the process of IL-6 down-regulation. Neutralization by an anti-IL-10 polyclonal antibody prevented IL-6 down-regulation in P. gingivalis LPS-pretreated monocytes, whereas IL-8 production was not affected. Addition of exogenous IL-10 during the high-dose LPS stimulation of untreated cells substituted for the LPS pretreatment and resulted in the inhibition of IL-6 production in a dose-dependent manner. A higher dose of IL-10 was required to suppress IL-8 synthesis from monocytes. Our data suggest that IL-10 mediates IL-6 down-regulation in P. gingivalis LPS-tolerant monocytes in an autocrine manner.

Abstract 235: Atorvastatin Has No Impact on the Effects of CETP Inhibitors on HDL Structure and Cholesterol Efflux Capacity in Chow-Fed Rabbits

2014 ◽  
Vol 34 (suppl_1) ◽  
Author(s):  
Mathieu R Brodeur ◽  
David Rhainds ◽  
Daniel Charpentier ◽  
Téodora Mihalache-Avram ◽  
Mélanie Mecteau ◽  
...  
Keyword(s):  
Cholesterol Efflux ◽  
Mononuclear Cells ◽  
Denaturing Gradient Gel Electrophoresis ◽  
Human Peripheral Blood ◽  
Cell Types ◽  
Peripheral Blood Mononuclear ◽  
Cholesterol Efflux Capacity ◽  
Cetp Inhibitors ◽  
Cetp Inhibition ◽  
The Impact

Introduction: In the dal-OUTCOMES trial, cholesteryl ester transfer protein (CETP) inhibition by dalcetrapib increased HDL-C but had a neutral effect on CV risk. However, these patients were also treated with standard-of-care therapies including statins, a drug class shown to reduce ABCA1 mRNA expression in human peripheral blood mononuclear cells (PBMC). Hypothesis: Given the role of ABCA1 in cholesterol efflux and HDL biogenesis, we determined the impact of atorvastatin on the potential beneficial effects provided by dalcetrapib and anacetrapib, two CETP inhibitors. Methods: Dalcetrapib (300 mg/kg) or anacetrapib (30 mg/kg) were administered to New Zealand White rabbits receiving or not atorvastatin (2.5 mg/kg) for 14 days. Lipid profiles were measured biochemically. ApoA-I distribution in HDL subclasses was evaluated by 1D-non-denaturing gradient gel electrophoresis (1D-NDGGE). BHK cells with inducible expression of human ABCA1 and HepG2 hepatocytes were used to measure ABCA1- and SR-BI-dépendent cholesterol efflux. Human THP-1 macrophages and PBMC isolated from rabbits were also used for efflux assays. Results: Dalcetrapib increased HDL-C by +81% (p<0.01) and total apoA-I (1D-NDGGE) by 1.5-fold (p<0.01). This was associated with an increase in both large and small α-migrating HDL (+52%, p<0.05; +74%, p<0.05). Cholesterol efflux showed that ABCA1- and SR-BI-dependent effluxes to apoB-depleted serum were increased in dalcetrapib-treated rabbits (+31%, p<0.01; +38%, p<0.001). Atorvastatin had no impact on all these parameters. Moreover, treatment of THP-1 with atorvastatin did not inhibit the increase of cholesterol efflux (+24%, p<0.01) induced by depleted serum from dalcetrapib-treated rabbits. Efflux capacity of PBMC isolated from rabbits treated with atorvastatin was also unchanged compared to the other groups. The absence of effect of atorvastatin on cholesterol efflux was also observed with anacetrapib. Conclusion: CETP inhibition increased HDL-C, apoA-I levels and cholesterol efflux from different cell types. Atorvastatin had no effect on these parameters, suggesting that the increased cholesterol efflux capacity induced by CETPi is not hampered by statins.

scholarly journals Moroccan Propolis: A Natural Antioxidant, Antibacterial, and Antibiofilm against Staphylococcus aureus with No Induction of Resistance after Continuous Exposure

2018 ◽  
Vol 2018 ◽  
pp. 1-19 ◽  
Author(s):  
Soukaïna El-Guendouz ◽  
Smail Aazza ◽  
Badiaa Lyoussi ◽  
Vassya Bankova ◽  
Milena Popova ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Quorum Sensing ◽  
Galleria Mellonella ◽  
Antioxidant Activities ◽  
Bacterial Biofilm ◽  
Continuous Exposure ◽  
Dependent Manner ◽  
Propolis Extract ◽  
Two Samples ◽  
The Impact

This study was performed to evaluate the total phenols, flavonoids, and antioxidant activities of twenty-four propolis samples from different regions of Morocco. In addition, two samples were screened regarding the antibacterial effect against four Staphylococcus aureus strains. Gas chromatography coupled to mass spectra (GC-MS) analysis was done for propolis samples used in antibacterial tests. The minimum inhibitory and minimum bactericidal concentration (MIC, MBC) were determined. The potential to acquire the resistance after sequential exposure of bacterial strains and the impact of adaptation to propolis on virulence using the Galleria mellonella were evaluated. Additionally, the effects of propolis extract on the bacterial adherence ability and its ability to inhibit the quorum sensing activity were also examined. Among the twenty-four extracts studied, the samples from Sefrou, Outat el Haj, and the two samples marketed in Morocco were the best for scavenging DPPH, ABTS, NO, peroxyl, and superoxide radicals as well as in scavenging of hydrogen peroxide. A strong correlation was found between the amounts of phenols, flavonoids, and antioxidant activities. Propolis extract at the MIC value (0.36 mg/mL) significantly reduced (p < 0.001) the virulence potential of S. aureus ATCC 6538 and the MRSA strains without leading to the development of resistance in the sequence of continuous exposure. It was able to impair the bacterial biofilm formation. The results have revealed that sample 1 reduces violacein production in a concentration dependent manner, indicating inhibition of quorum sensing. This extract has as main group of secondary metabolites flavonoids (31.9%), diterpenes (21.5%), and phenolic acid esters (16.5%).

scholarly journals Differentiation stimuli induce receptors for plasma fibronectin on the human myelomonocytic cell line HL-60

Blood ◽  
1984 ◽  
Vol 64 (4) ◽  
pp. 858-866 ◽  
Author(s):  
CG Pommier ◽  
J O'Shea ◽  
T Chused ◽  
T Takahashi ◽  
M Ochoa ◽  
...  
Keyword(s):  
Cell Line ◽  
Human Peripheral Blood ◽  
Cell Types ◽  
Plasma Fibronectin ◽  
Blood Monocytes ◽  
Before And After ◽  
Differentiating Agents ◽  
Membrane Antigens ◽  
Human Polymorphonuclear Leukocytes

Abstract Plasma fibronectin (Fn) induces phagocytosis of C3b-opsonized sheep erythrocytes (EC3b) by human peripheral blood monocytes. However, Fn does not induce erythrophagocytosis of EC3b by human polymorphonuclear leukocytes (PMN), unless the PMN have been exposed to C5a or N-formyl- methionyl-leucyl-phenylalanine. Because of this difference, it is of great interest to examine Fn binding to cells that possess the capacity to differentiate into either granulocytes or monocytes. Hence, we have examined the consequences of Fn binding to the human myelomonocytic cell line, HL-60, both before and after in vitro differentiation of the HL-60, along a monocytoid or a granulocytoid pathway. Fn receptors were not found on undifferentiated HL-60, but several differentiating agents promoted the HL-60 binding of Fn-coated microspheres (Fn-ms). The peak of Fn-ms binding occurred four to five days after the induction of differentiation with dimethylsulfoxide (DMSO), and two days after induction by PMA. In addition, cells that differentiated along either the monocytoid or the granulocytoid pathway showed a marked increase in the phagocytosis of both IgG-coated erythrocytes (EA) and EC3b when they were exposed to Fn. Comparison of the effects of anti-Fn monoclonals on the binding of Fn-ms to the monocytes, PMN, and HL-60 showed that the same monoclonals block Fn-ms-binding and Fn-induced EC3b phagocytosis by all three cell types. Two monoclonal antibodies, M1/70 and A6F10, directed against membrane antigens on PMN and monocytes, inhibited Fn-ms binding. Both also blocked Fn-induced EC3b ingestion by these cells. However, neither antibody blocked Fn-ms binding or EC3b ingestion by differentiated HL-60. We conclude that differentiated HL-60 cells express functionally active Fn receptors, similar to monocytes and activated PMN, which, nonetheless, differ from normal cells in their association with the antigens recognized by M1/70 and A6F10.

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