scholarly journals In Vitro Effects of the Reduced Form of Coenzyme Q10 on Secretion Levels of TNF-α and Chemokines in Response to LPS in the Human Monocytic Cell Line THP-1

2009 ◽  
Vol 44 (1) ◽  
pp. 62-66 ◽  
Author(s):  
Constance Schmelzer ◽  
Gerti Lorenz ◽  
Gerald Rimbach ◽  
Frank Döring
Keyword(s):  
Cell Line ◽  
Coenzyme Q10 ◽  
Reduced Form ◽  
Monocytic Cell ◽  
Monocytic Cell Line ◽  
Human Monocytic Cell Line ◽  
Tnf Α ◽  
Human Monocytic Cell ◽  
In Vitro Effects

scholarly journals Alkaline Comet Assay using the monocytic cell line THP-1

2019 ◽  
Author(s):  
Ana Neves-Costa ◽  
Dora Pedroso ◽  
Luis F Moita
Keyword(s):  
Comet Assay ◽  
Cell Line ◽  
Adherent Cell ◽  
Monocytic Cell ◽  
Monocytic Cell Line ◽  
Strand Breaks ◽  
Human Monocytic Cell Line ◽  
Human Monocytic Cell ◽  
Dna Strand

Abstract This protocol details the experimental procedure for performing the comet assay, a very sensitive DNA break assay based on single cell gel electrophoresis.The analysis of DNA strand breaks, both single- and double-strand breaks (SSBs and DSBs, respectively), was performed in immune responsive cells. The cell line used was the human monocytic cell line THP-1, an adherent cell type with many known applications in in vitro studies of innate immunity. The comet assay is a robust procedure that allows the accurate and reproducible quantification of DNA damage. Here we describe not only the comet assay step-by-step protocol, but also some important aspects related to troubleshooting.

scholarly journals Alkaline Comet Assay using the monocytic cell line THP-1

2019 ◽  
Author(s):  
Ana Neves-Costa ◽  
Dora Pedroso ◽  
Luis F Moita
Keyword(s):  
Comet Assay ◽  
Cell Line ◽  
Adherent Cell ◽  
Monocytic Cell ◽  
Monocytic Cell Line ◽  
Strand Breaks ◽  
Human Monocytic Cell Line ◽  
Human Monocytic Cell ◽  
Dna Strand

Abstract This protocol details the experimental procedure for performing the comet assay, a very sensitive DNA break assay based on single cell gel electrophoresis.The analysis of DNA strand breaks, both single- and double-strand breaks (SSBs and DSBs, respectively), was performed in immune responsive cells. The cell line used was the human monocytic cell line THP-1, an adherent cell type with many known applications in in vitro studies of innate immunity. The comet assay is a robust procedure that allows the accurate and reproducible quantification of DNA damage. Here we describe not only the comet assay step-by-step protocol, but also some important aspects related to troubleshooting.

scholarly journals Regulation of Proinflammatory Cytokine Expression by Shiga Toxin 1 and/or Lipopolysaccharides in the Human Monocytic Cell Line THP-1

2004 ◽  
Vol 72 (5) ◽  
pp. 2618-2627 ◽  
Author(s):  
Lisa M. Harrison ◽  
Wilhelmina C. E. van Haaften ◽  
Vernon L. Tesh
Keyword(s):  
Cell Line ◽  
Shiga Toxin ◽  
Cytokine Expression ◽  
Receptor Expression ◽  
Monocytic Cell ◽  
Monocytic Cell Line ◽  
Human Monocytic Cell Line ◽  
Mrna Induction ◽  
Tnf Α ◽  
Human Monocytic Cell

ABSTRACT Infection with Shiga toxin (Stx)-producing bacteria and the subsequent release of Stxs and endotoxins into the bloodstream may damage blood vessels in the colon, kidneys, and central nervous system, leading to bloody diarrhea, acute renal failure, and neurological complications. The proinflammatory cytokines tumor necrosis factor alpha (TNF-α) and interleukin-1β (IL-1β) may contribute to the pathogenesis of Stx-induced vascular lesions by up-regulating toxin receptor expression on endothelial cells. We previously showed that macrophages treated with purified Shiga toxin 1 (Stx1) or lipopolysaccharides (LPS) secrete TNF-α and IL-1β. Northern blot analysis revealed that treatment of the human monocytic cell line THP-1 with LPS induced a rapid and transient increase in steady-state TNF-α and IL-1β transcripts. In contrast, Stx1 induced slower but prolonged elevations in cytokine transcripts. The presence of both stimulants resulted in optimal cytokine mRNA induction in terms of kinetics and prolonged expression. Compared to LPS, Stx1 was a poor inducer of IL-1β protein expression, although levels of soluble IL-1β induced by all treatments continually increased over 72 h. IL-1β transcripts were not induced by Stx1 B-subunits. Using the transcriptional inhibitor actinomycin D, we determined that treatment with Stx1 or Stx1 plus LPS induced cytokine transcripts with increased stability compared to transcripts induced by LPS alone. For all treatments, IL-1β mRNA decay was slower than TNF-α. Collectively, our data suggest that Stxs affect cytokine expression, in part, at the posttranscriptional level by stabilizing mRNAs. Optimal TNF-α expression occurs when both Stxs and LPS are present.

scholarly journals Bioengineered Nanoparticles Loaded-Hydrogels to Target TNF Alpha in Inflammatory Diseases

Pharmaceutics ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1111
Author(s):  
Isabel Matos Oliveira ◽  
Diogo Castro Fernandes ◽  
Fátima Raquel Maia ◽  
Raphael Faustino Canadas ◽  
Rui Luís Reis ◽  
...  
Keyword(s):  
Inflammatory Diseases ◽  
Gellan Gum ◽  
Monocytic Cell ◽  
Monocytic Cell Line ◽  
Human Monocytic Cell Line ◽  
Tnf Α ◽  
Human Monocytic Cell ◽  
Vitro Model ◽  
Static Conditions

Rheumatoid Arthritis (RA) is an incurable autoimmune disease that promotes the chronic impairment of patients’ mobility. For this reason, it is vital to develop therapies that target early inflammatory symptoms and act before permanent articular damage. The present study offers two novel therapies based in advanced drug delivery systems for RA treatment: encapsulated chondroitin sulfate modified poly(amidoamine) dendrimer nanoparticles (NPs) covalently bonded to monoclonal anti-TNF α antibody in both Tyramine-Gellan Gum and Tyramine-Gellan Gum/Silk Fibroin hydrogels. Using pro-inflammatory THP-1 (i.e., human monocytic cell line), the therapy was tested in an inflammation in vitro model under both static and dynamic conditions. Firstly, we demonstrated effective NP-antibody functionalization and TNF-α capture. Upon encapsulation, the NPs were released steadily over 21 days. Moreover, in static conditions, the approaches presented good anti-inflammatory activity over time, enabling the retainment of a high percentage of TNF α. To mimic the physiological conditions of the human body, the hydrogels were evaluated in a dual-chamber bioreactor. Dynamic in vitro studies showed absent cytotoxicity in THP-1 cells and a significant reduction of TNF-α in suspension over 14 days for both hydrogels. Thus, the developed approach showed potential for use as personalized medicine to obtain better therapeutic outcomes and decreased adverse effects.

scholarly journals Production of erythroid potentiating factor(s) by a human monocytic cell line

Blood ◽  
1981 ◽  
Vol 57 (1) ◽  
pp. 170-173 ◽  
Author(s):  
JL Ascensao ◽  
NE Kay ◽  
T Earenfight-Engler ◽  
HS Koren ◽  
ED Zanjani
Keyword(s):  
Cell Line ◽  
Conditioned Medium ◽  
Human Cell Line ◽  
Monocytic Cell ◽  
Monocytic Cell Line ◽  
Serum Free ◽  
Human Monocytic Cell Line ◽  
Human Monocytic Cell ◽  
Dose Dependent

Abstract U-937 is a human monocytic cell line that has been to elaborate factors that affect normal human hematopoiesis in vitro. Studies on the effects of these factors demonstrated an erythroid potentiating factor (EPF) and a potent inhibitor of granulocyte-macrophage (CFU-GM) colony growth. The EPF was present in both serum-containing and serum-free U- 937 conditioned media, had a dose-dependent effect on erythroid colony formation and was remarkably heart stable. The CFU-GM inhibitory activity was also detected in serum-free conditioned medium, was dose- dependent, heart labile and its effect was reversed by Indomethacin. Indomethacin (Sigma, St. Louis, Mo.) did not alter the erythroid effects of the U-937 conditioned medium. No colony stimulating factor (CSF) or erythropoietin (Ep) could be detected in this medium. The existence of a human cell line capable of production EPF without simultaneous CSF production will permit further studies on the biochemical and biologic nature of these factors.

scholarly journals Production of erythroid potentiating factor(s) by a human monocytic cell line

Blood ◽  
1981 ◽  
Vol 57 (1) ◽  
pp. 170-173
Author(s):  
JL Ascensao ◽  
NE Kay ◽  
T Earenfight-Engler ◽  
HS Koren ◽  
ED Zanjani
Keyword(s):  
Cell Line ◽  
Conditioned Medium ◽  
Human Cell Line ◽  
Monocytic Cell ◽  
Monocytic Cell Line ◽  
Serum Free ◽  
Human Monocytic Cell Line ◽  
Human Monocytic Cell ◽  
Dose Dependent

U-937 is a human monocytic cell line that has been to elaborate factors that affect normal human hematopoiesis in vitro. Studies on the effects of these factors demonstrated an erythroid potentiating factor (EPF) and a potent inhibitor of granulocyte-macrophage (CFU-GM) colony growth. The EPF was present in both serum-containing and serum-free U- 937 conditioned media, had a dose-dependent effect on erythroid colony formation and was remarkably heart stable. The CFU-GM inhibitory activity was also detected in serum-free conditioned medium, was dose- dependent, heart labile and its effect was reversed by Indomethacin. Indomethacin (Sigma, St. Louis, Mo.) did not alter the erythroid effects of the U-937 conditioned medium. No colony stimulating factor (CSF) or erythropoietin (Ep) could be detected in this medium. The existence of a human cell line capable of production EPF without simultaneous CSF production will permit further studies on the biochemical and biologic nature of these factors.

scholarly journals Interleukin-10 Modulates Proinflammatory Cytokines in the Human Monocytic Cell Line THP-1 Stimulated withBorrelia burgdorferi Lipoproteins

2000 ◽  
Vol 68 (12) ◽  
pp. 6663-6669 ◽  
Author(s):  
P. K. Murthy ◽  
Vida A. Dennis ◽  
Barbara L. Lasater ◽  
Mario T. Philipp
Keyword(s):  
Cell Line ◽  
Inflammatory Cytokines ◽  
Cytokine Production ◽  
Interleukin 10 ◽  
Monocytic Cell ◽  
Monocytic Cell Line ◽  
Human Monocytic Cell Line ◽  
Tnf Α ◽  
Human Monocytic Cell ◽  
Kinetics Of

ABSTRACT We determined previously that lipoproteins of Borrelia burgdorferi stimulate inflammatory and anti-inflammatory cytokines (interleukin-10 [IL-10]) in monocytes. IL-10 could have an effect on innate and acquired immune responses to B. burgdorferi and influence the magnitude of the infectious inoculum and disease outcome. To understand the mechanism(s) of IL-10 action during early infection, when innate immunity expressed chiefly by skin macrophages is key, we investigated the effect of exogenous and endogenous IL-10 on the production of the macrophage-derived cytokines IL-6, IL-1β, IL-12, and tumor necrosis factor alpha (TNF-α). We used the THP-1 human monocytic cell line and recombinant lipidated OspA (L-OspA) as the model target cell and stimulant, respectively. To determine the kinetics of cytokine production by THP-1 cells, we stimulated them with L-OspA and also with heat-killed B. burgdorferi cells (HBb) and lipopolysaccharide (LPS). Exogenous IL-10 dampened production of inflammatory cytokines, as elicited by lipoproteins. The inhibition of endogenous IL-10 function by anti-IL-10 antibody reduced the production of IL-12 and IL-6 but not that of IL-1β and TNF-α. An inspection of the kinetics of cytokine production clarified this finding. TNF-α was produced prior to, and IL-β was produced at the same time as, IL-10, whereas IL-6 and IL-12 were produced later. HBb, LPS, and L-OspA yielded similar kinetics of cytokine production. This result reinforces the notion that lipoproteins are the functional molecules in HBb and perhaps in vivo. It indicates also that signaling pathways utilized by LPS and lipoproteins may be extensively shared. The results are consistent with a major role played by IL-10 in controlling the initial phase of infection with this spirochete.

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