Transcriptomic Profiling of the Development of the Inflammatory Response in Human Monocytes In Vitro

SummaryHuman monocytes in vitro respond to various agents (immune complexes, lectins, endotoxin, the divalent ionophore A 23187, 12-0-tetradecanoyl-phorbol 13-acetate [TPA], purified protein derivative [PPD] of Bacille Calmette-Guerin) with an increased synthesis of the protein component of thromboplastin. The effect of cyclic AMP and cyclic GMP on this response has been studied. Dibutyryl-cyclic AMP, prostaglandin E1 and the phosphodiesterase inhibitors 3-butyl-1-methyl-xanthine (MIX) and rac -4-(3-butoxy-4-methoxybenzyl)-2-imidazolidinone (Ro 201724), separately and in combination have a pronounced inhibitory effect on the response to immune complexes and PPD, and a moderate effect on the response to endotoxin and lectins. The effect on TPA response and on the response to A 23187 was slight. Dibutyryl-cyclic GMP (1 mM) gave a slight inhibition of the TPA arid IC response, but had essentially no effect on the response to other inducers. The intracellular cAMP level increased when monocytes were incubated with IC, TPA or A 23187 followed by a decrease to basal levels within 1-2 hr, whereas lectin (PHA) and PPD did not induce such changes. The cAMP response to endotoxin varied. Stimulation with IC induced an increase in monocyte cGMP levels, whereas the other stimulants did not cause such changes.

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Twin studies demonstrate a host cell genetic effect on productive human immunodeficiency virus infection of human monocytes and macrophages in vitro.

Journal of Virology ◽

10.1128/jvi.70.11.7792-7803.1996 ◽

1996 ◽

Vol 70 (11) ◽

pp. 7792-7803 ◽

Cited By ~ 36

Author(s):

J Chang ◽

H M Naif ◽

S Li ◽

J S Sullivan ◽

C M Randle ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Virus Infection ◽

Human Immunodeficiency Virus Infection ◽

Twin Studies ◽

Genetic Effect ◽

Human Monocytes ◽

Immunodeficiency Virus ◽

Monocytes And Macrophages ◽

Human Monocytes And Macrophages

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Macrophagic Inflammatory Response Next to Dental Implants with Different Macro- and Micro-Structure: An In Vitro Study

Applied Sciences ◽

10.3390/app11125324 ◽

2021 ◽

Vol 11 (12) ◽

pp. 5324

Author(s):

Maria Menini ◽

Francesca Delucchi ◽

Domenico Baldi ◽

Francesco Pera ◽

Francesco Bagnasco ◽

...

Keyword(s):

Inflammatory Response ◽

Dental Implants ◽

In Vitro Study ◽

Titanium Implants ◽

Implant Surface ◽

Bone Implant ◽

Optimal Outcomes ◽

Negative Controls ◽

Inflammatory Effect

(1) Background: Intrinsic characteristics of the implant surface and the possible presence of endotoxins may affect the bone–implant interface and cause an inflammatory response. This study aims to evaluate the possible inflammatory response induced in vitro in macrophages in contact with five different commercially available dental implants. (2) Methods: one zirconia implant NobelPearl® (Nobel Biocare) and four titanium implants, Syra® (Sweden & Martina), Prama® (Sweden & Martina), 3iT3® (Biomet 3i) and Shard® (Mech & Human), were evaluated. After 4 h of contact of murine macrophage cells J774a.1 with the implants, the total RNA was extracted, transcribed to cDNA and the gene expression of the macrophages was evaluated by quantitative PCR (qPCR) in relation to the following genes: GAPDH, YWHAZ, IL1β, IL6, TNFα, NOS2, MMP-9, MMP-8 and TIMP3. The results were statistically analyzed and compared with negative controls. (3) Results: No implant triggered a significant inflammatory response in macrophages, although 3iT3 exhibited a slight pro-inflammatory effect compared to other samples. (4) Conclusions: All the samples showed optimal outcomes without any inflammatory stimulus on the examined macrophagic cells.

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Lipopolysaccharide induces neuroinflammation in microglia by activating the MTOR pathway and downregulating Vps34 to inhibit autophagosome formation

Journal of Neuroinflammation ◽

10.1186/s12974-019-1644-8 ◽

2020 ◽

Vol 17 (1) ◽

Cited By ~ 8

Author(s):

Xiaoxia Ye ◽

Mingming Zhu ◽

Xiaohang Che ◽

Huiyang Wang ◽

Xing-Jie Liang ◽

...

Keyword(s):

Inflammatory Response ◽

Microglial Activation ◽

Adaptor Protein ◽

Mtor Pathway ◽

Microglial Cells ◽

Inflammatory Factors ◽

Intraventricular Injection ◽

Enzyme Linked Immunosorbent Assay ◽

Autophagic Flux

Abstract Background Microglial activation is a prominent feature of neuroinflammation, which is present in almost all neurodegenerative diseases. While an initial inflammatory response mediated by microglia is considered to be protective, excessive pro-inflammatory response of microglia contributes to the pathogenesis of neurodegeneration. Although autophagy is involved in the suppression of inflammation, its role and mechanism in microglia are unclear. Methods In the present study, we studied the mechanism by which lipopolysaccharide (LPS) affects microglial autophagy and the effects of autophagy on the production of pro-inflammatory factors in microglial cells by western blotting, immunocytochemistry, transfection, transmission electron microscopy (TEM), and real-time PCR. In a mouse model of neuroinflammation, generated by intraventricular injection of LPS (5 μg/animal), we induced autophagy by rapamycin injection and investigated the effects of enhanced autophagy on microglial activation by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry. Results We found that autophagic flux was suppressed in LPS-stimulated N9 microglial cells, as evidenced by decreased expression of the autophagy marker LC3-II (lipidated form of MAP1LC3), as well as increased levels of the autophagy adaptor protein SQSTM1. LPS significantly decreased Vps34 expression in N9 microglial cells by activating the PI3KI/AKT/MTOR pathway without affecting the levels of lysosome-associated proteins and enzymes. More importantly, overexpression of Vps34 significantly enhanced the autophagic flux and decreased the accumulation of SQSTM1 in LPS-stimulated N9 microglial cells. Moreover, our results revealed that an LPS-induced reduction in the level of Vps34 prevented the maturation of omegasomes to phagophores. Furthermore, LPS-induced neuroinflammation was significantly ameliorated by treatment with the autophagy inducer rapamycin both in vitro and in vivo. Conclusions These data reveal that LPS-induced neuroinflammation in N9 microglial cells is associated with the inhibition of autophagic flux through the activation of the PI3KI/AKT/MTOR pathway, while enhanced microglial autophagy downregulates LPS-induced neuroinflammation. Thus, this study suggests that promoting the early stages of autophagy might be a potential therapeutic approach for neuroinflammation-associated diseases.

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In vitro knockout of human p47phox blocks superoxide anion production and LDL oxidation by activated human monocytes

The Journal of Lipid Research ◽

10.1016/s0022-2275(20)34488-6 ◽

2000 ◽

Vol 41 (3) ◽

pp. 489-495

Author(s):

Erik A. Bey ◽

Martha K. Cathcart

Keyword(s):

Superoxide Anion ◽

Ldl Oxidation ◽

Human Monocytes ◽

Superoxide Anion Production ◽

Anion Production

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Long non-coding RNA OIP5-AS1 aggravates acute lung injury by promoting inflammation and cell apoptosis via regulating the miR-26a-5p/TLR4 axis

BMC Pulmonary Medicine ◽

10.1186/s12890-021-01589-1 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Qingsong Sun ◽

Man Luo ◽

Zhiwei Gao ◽

Xiang Han ◽

Weiqin Wu ◽

...

Keyword(s):

Acute Lung Injury ◽

Lung Injury ◽

Inflammatory Response ◽

Cell Apoptosis ◽

Quantitative Polymerase Chain Reaction ◽

Luciferase Reporter ◽

Enzyme Linked Immunosorbent Assay ◽

Interacting Protein ◽

Wide Range

Abstract Background Acute lung injury (ALI) is a pulmonary disorder that leads to acute respiration failure and thereby results in a high mortality worldwide. Increasing studies have indicated that toll-like receptor 4 (TLR4) is a promoter in ALI, and we aimed to explore the underlying upstream mechanism of TLR4 in ALI. Methods We used lipopolysaccharide (LPS) to induce an acute inflammatory response in vitro model and a murine mouse model. A wide range of experiments including reverse transcription quantitative polymerase chain reaction, western blot, enzyme linked immunosorbent assay, flow cytometry, hematoxylin–eosin staining, RNA immunoprecipitation, luciferase activity and caspase-3 activity detection assays were conducted to figure out the expression status, specific role and potential upstream mechanism of TLR4 in ALI. Result TLR4 expression was upregulated in ALI mice and LPS-treated primary bronchial/tracheal epithelial cells. Moreover, miR-26a-5p was confirmed to target TLR4 according to results of luciferase reporter assay. In addition, miR-26a-5p overexpression decreased the contents of proinflammatory factors and inhibited cell apoptosis, while upregulation of TLR4 reversed these effects of miR-26a-5p mimics, implying that miR-26a-5p alleviated ALI by regulating TLR4. Afterwards, OPA interacting protein 5 antisense RNA 1 (OIP5-AS1) was identified to bind with miR-26a-5p. Functionally, OIP5-AS1 upregulation promoted the inflammation and miR-26a-5p overexpression counteracted the influence of OIP5-AS1 upregulation on cell inflammatory response and apoptosis. Conclusion OIP5-AS1 promotes ALI by regulating the miR-26a-5p/TLR4 axis in ALI mice and LPS-treated cells, which indicates a promising insight into diagnostics and therapeutics in ALI.

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Reduced Ex Vivo Interleukin-8 Production by Neutrophils in Septic and Nonseptic Systemic Inflammatory Response Syndrome

Blood ◽

10.1182/blood.v91.9.3439 ◽

1998 ◽

Vol 91 (9) ◽

pp. 3439-3446 ◽

Cited By ~ 79

Author(s):

Christelle Marie ◽

Jane Muret ◽

Catherine Fitting ◽

Marie-Reine Losser ◽

Didier Payen ◽

...

Keyword(s):

Systemic Inflammatory Response Syndrome ◽

Inflammatory Response ◽

Ex Vivo ◽

Endotoxin Tolerance ◽

Systemic Inflammatory Response ◽

Interleukin 8 ◽

Polymorphonuclear Cells ◽

Healthy Controls ◽

Relative Contribution

AbstractEx vivo cytokine production by circulating lymphocytes and monocytes is reduced in patients with infectious or noninfectious systemic inflammatory response syndrome. Very few studies have addressed the reactivity of polymorphonuclear cells (PMN). To analyze further the relative contribution of systemic inflammatory response syndrome alone or in combination with infection we studied the interleukin-8 (IL-8) production by PMN isolated from patients who had undergone cardiac surgery with cardiopulmonary bypass (CPB) and patients with sepsis. Cells were activated with either lipopolysaccharide (LPS) or heat-killed streptococci. Compared with healthy controls, the release of IL-8 by PMN in both groups of patients was significantly reduced whether activated by LPS, independently of its concentration and origin, or by heat-killed streptococci. These observations suggest that stressful conditions related to inflammation, independently of infection, rapidly dampened the reactivity of circulating PMN. We investigated whether the observed diminished reactivity of PMN might reflect an endotoxin tolerance phenomenon. Our in vitro experiments with PMN from healthy controls indicated that PMN could not be rendered tolerant stricto sensu. However, our data suggested that LPS-induced mediators such as IL-10 may be responsible for the observed anergy in patients.

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