scholarly journals Neisseria gonorrhoeaeInduces a Tolerogenic Phenotype in Macrophages to Modulate Host Immunity

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Alejandro Escobar ◽  
Enzo Candia ◽  
Sebastian Reyes-Cerpa ◽  
Bélgica Villegas-Valdes ◽  
Tanya Neira ◽  
...  
Keyword(s):  
Immune Response ◽  
Cell Line ◽  
Mhc Class Ii ◽  
Transmitted Disease ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Sexually Transmitted ◽  
Antigen Presenting

Neisseria gonorrhoeaeis the etiological agent of gonorrhoea, which is a sexually transmitted disease widespread throughout the world.N. gonorrhoeaedoes not improve immune response in patients with reinfection, suggesting that gonococcus displays several mechanisms to evade immune response and survive in the host.N. gonorrhoeaeis able to suppress the protective immune response at different levels, such as B and T lymphocytes and dendritic cells. In this study, we determined whetherN. gonorrhoeaedirectly conditions the phenotype of RAW 264.7 murine macrophage cell line and its response. We established that gonococcus was effectively phagocytosed by the RAW 264.7 cells and upregulates production of immunoregulatory cytokines (IL-10 and TGF-β1) but not the production of proinflammatory cytokine TNF-α, indicating that gonococcus induces a shift towards anti-inflammatory cytokine production. Moreover,N. gonorrhoeaedid not induce significant upregulation of costimulatory CD86 and MHC class II molecules. We also showed thatN. gonorrhoeaeinfected macrophage cell line fails to elicit proliferative CD4+ response. This implies that macrophage that can phagocytose gonococcus do not display proper antigen-presenting functions. These results indicate thatN. gonorrhoeaeinduces a tolerogenic phenotype in antigen-presenting cells, which seems to be one of the mechanisms to induce evasion of immune response.

scholarly journals Down-Regulation of Glycosylphosphatidylinositol-Specific Phospholipase D Induced by Lipopolysaccharide and Oxidative Stress in the Murine Monocyte- Macrophage Cell Line RAW 264.7

2001 ◽  
Vol 69 (5) ◽  
pp. 3214-3223 ◽  
Author(s):  
Xiaohan Du ◽  
Martin G. Low
Keyword(s):  
Oxidative Stress ◽  
Inflammatory Response ◽  
Cell Line ◽  
Phospholipase D ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Down Regulation ◽  
Tnf Α

ABSTRACT Serum glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD) activity is reduced over 75% in systemic inflammatory response syndrome. To investigate the mechanism of this response, expression of the GPI-PLD gene was studied in the mouse monocyte-macrophage cell line RAW 264.7 stimulated with lipopolysaccharide (LPS; 0.5 to 50 ng/ml). GPI-PLD mRNA was reduced approximately 60% in a time- and dose-dependent manner. Oxidative stress induced by 0.5 mM H2O2 or 50 μM menadione also caused a greater than 50% reduction in GPI-PLD mRNA. The antioxidant N-acetyl-l-cysteine attenuated the down-regulatory effect of H2O2but not of LPS. Cotreatment of the cells with actinomycin D inhibited down-regulation induced by either LPS or H2O2. The half-life of GPI-PLD mRNA was not affected by LPS, or decreased slightly with H2O2, indicating that the reduction in GPI-PLD mRNA is due primarily to transcriptional regulation. Stimulation with tumor necrosis factor alpha (TNF-α) resulted in ∼40% reduction in GPI-PLD mRNA in human A549 alveolar carcinoma cells but not RAW 264.7 cells, suggesting that alternative pathways could exist in different cell types for down-regulating GPI-PLD expression during an inflammatory response and the TNF-α autocrine signaling mechanism alone is not sufficient to recapitulate the LPS-induced reduction of GPI-PLD in macrophages. Sublines of RAW 264.7 cells with reduced GPI-PLD expression exhibited increased cell sensitivity to LPS stimulation and membrane-anchored CD14 expression on the cell surface. Our data suggest that down-regulation of GPI-PLD could play an important role in the control of proinflammatory responses.

Ferritin does not donate its iron for haem synthesis in macrophages

2010 ◽  
Vol 429 (3) ◽  
pp. 463-471 ◽  
Author(s):  
Marc Mikhael ◽  
Alex D. Sheftel ◽  
Prem Ponka
Keyword(s):  
Single Cell ◽  
Cell Line ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Murine Macrophage ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Iron Storage ◽  
Subunit Protein ◽  
Metabolic Functions

Iron is essential for all life, yet can be dangerous under certain conditions. Iron storage by the 24-subunit protein ferritin renders excess amounts of the metal non-reactive and, consequentially, ferritin is crucial for life. Although the mechanism detailing the storage of iron in ferritin has been well characterized, little is known about the fate of ferritin-stored iron and whether it can be released and reutilized for metabolic use within a single cell. Virtually nothing is known about the use of ferritin-derived iron in non-erythroid cells. We therefore attempted to answer the question of whether iron from ferritin can be used for haem synthesis in the murine macrophage cell line RAW 264.7 cells. Cells treated with ALA (5-aminolaevulinic acid; a precursor of haem synthesis) show increased haem production as determined by enhanced incorporation of transferrin-bound 59Fe into haem. However, the present study shows that, upon the addition of ALA, 59Fe from ferritin cannot be incorporated into haem. Additionally, little 59Fe is liberated from ferritin when haem synthesis is increased upon addition of ALA. In conclusion, ferritin in cultivated macrophages is not a significant source of iron for the cell's own metabolic functions.

scholarly journals Dicalcium Silicate Induced Proinflammatory Responses through TLR2-Mediated NF-κB and JNK Pathways in the Murine RAW 264.7 Macrophage Cell Line

2018 ◽  
Vol 2018 ◽  
pp. 1-16 ◽  
Author(s):  
Shixiang Lai ◽  
Liangjiao Chen ◽  
Wei Cao ◽  
Shiman Cui ◽  
Xingyang Li ◽  
...  
Keyword(s):  
Cell Line ◽  
Dicalcium Silicate ◽  
Cell Counting ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Proinflammatory Response ◽  
Raw 264.7 Macrophage ◽  
Proinflammatory Responses

Proinflammatory responses are important aspects of the immune response to biomaterials, which may cause peri-implantitis and implant shedding. The purpose of this study was to test the cytotoxicity and proinflammatory effects of dicalcium silicate particles on RAW 264.7 macrophages and to investigate the proinflammatory response mechanism induced by C2S and tricalcium phosphate (TCP). C2S and TCP particles were characterized using scanning electron microscopy (SEM), energy spectrum analysis (EDS) and X-ray diffraction (XRD). Cytotoxicity and apoptosis assays with C2S and TCP in the murine RAW 264.7 cell line were tested using the cell counting kit-8 (CCK-8) assay and flow cytometry (FCM). The detection results showed that C2S and TCP particles had no obvious toxicity in RAW 264.7 cells and did not cause obvious apoptosis, although they both caused an oxidative stress response by producing ROS when the concentrations were at 100 μg/mL. C2S particles are likely to induce a proinflammatory response by inducing high TLR2, TNF-α mRNA, TNF-α proinflammatory cytokine, p-IκB, and p-JNK1 + JNK2 + JNK3 expression levels. When we added siRNA-TLR2-1, a significant reduction was observed. These findings support the theory that C2S particles induce proinflammatory responses through the TLR2-mediated NF-κB and JNK pathways in the murine RAW 264.7 macrophage cell line.

scholarly journals Short Communication: Immunostimulatory effect of tempoyak (fermented durian) on inducing cytokine production (IL-6 and TNF-α) by RAW 264.7 cells

2018 ◽  
Vol 19 (1) ◽  
pp. 318-322
Author(s):  
SULVANIA SUSANTO ◽  
ANTON SUMARPO ◽  
ARLI ADITYA PARIKESIT ◽  
AGUS BUDIAWAN NARO PUTRA ◽  
ERI ISHIDA ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Cell Line ◽  
Short Communication ◽  
Cytokine Production ◽  
Water Extract ◽  
Raw 264.7 Cells ◽  
Fermented Foods ◽  
Raw 264.7 ◽  
Macrophage Cell Line ◽  
Macrophage Cell

Susanto S, Sumarpo A, Parikesit AA, Putra ABNN, Ishida E, Tabuchi K, Sugahara T. 2018. Short Communication: Immunostimulatory effect of tempoyak (fermented durian) on inducing cytokine production (IL-6 and TNF-α) by RAW 264.7 cells. Biodiversitas 19: 318-322. Indonesia is known to be home to various fermented foods with many-reported usage as potential sources of probiotics. Tempoyak (fermented durian) is among one of the Indonesian fermented foods that are rarely studied for its bioactivities. This study was conducted to evaluate the potential bioactivities of tempoyak, particularly the immunostimulatory aspects. Water extract of tempoyak was prepared by suspending the freeze-dried tempoyak sample in distilled water. Immunostimulatory activity of tempoyak water extract was evaluated using mouse macrophage cell line RAW 264.7. ELISA was used to screen cytokine productions (IL-6 and TNF-α) by RAW 264.7 cells following treatment with tempoyak water extract. In addition, real-time RT-PCR was also used to determine IL-6 and TNF-α mRNA expression. We showed that water extract of tempoyak exerts immunostimulatory effects towards RAW 264.7 cells. This was observed from the increased production of IL-6 and TNF-α in a dose-dependent manner. This was also supported by increased IL-6 and TNF-α mRNA expression. Our finding suggests that tempoyak has immunostimulatory effects towards murine macrophage cell line RAW 264.7. However, further studies are needed to identify the specific compounds responsible for inducing immunostimulatory effects.

Immune Enhancing Effect of Medicinal Herb Extracts on a RAW 264.7 Macrophage Cell Line

2012 ◽  
Vol 41 (11) ◽  
pp. 1521-1527 ◽  
Author(s):  
A-Reum Yu ◽  
Ho-Young Park ◽  
In-Wook Choi ◽  
Yong-Kon Park ◽  
Hee-Do Hong ◽  
...  
Keyword(s):  
Cell Line ◽  
Medicinal Herb ◽  
Raw 264.7 ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Enhancing Effect ◽  
Raw 264.7 Macrophage ◽  
Herb Extracts

scholarly journals Evaluation of Methods for Transient Transfection of a Murine Macrophage Cell Line, RAW 264.7

BioTechniques ◽  
1999 ◽  
Vol 27 (4) ◽  
pp. 824-832 ◽  
Author(s):  
C.D. Thompson ◽  
M.R. Frazier-Jessen ◽  
R. Rawat ◽  
R.P. Nordan ◽  
R.T. Brown
Keyword(s):  
Cell Line ◽  
Transient Transfection ◽  
Raw 264.7 ◽  
Murine Macrophage ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Murine Macrophage Cell Line ◽  
Evaluation Of Methods
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