Functional Association of FcɛRIγ With Arginine632 of Paired Immunoglobulin-Like Receptor (PIR)-A3 in Murine Macrophages

Blood ◽  
1999 ◽  
Vol 94 (5) ◽  
pp. 1790-1796 ◽  
Author(s):  
Lynn S. Taylor ◽  
Daniel W. McVicar
Keyword(s):  
Transmembrane Domain ◽  
Dependent Manner ◽  
Macrophage Cell Line ◽  
Inhibitory Pathway ◽  
Macrophage Cell ◽  
Complete Understanding ◽  
Chimeric Receptors ◽  
Extracellular Region ◽  
Activating Receptors ◽  
Ifn Γ

Abstract Paired immunoglobulin-like receptors (PIR) are expressed on B cells and macrophages and include inhibitory and putative activating receptors referred to as PIR-B and PIR-A, respectively. Although PIR-B’s inhibitory pathway has been described, it is unknown whether PIR-A receptors can deliver activation signals to macrophages, and if so, through what mechanism. Here we use chimeric receptors to address the mechanisms of PIR-A signaling. Cotransfection of chimeric receptors comprised of the extracellular region of human CD4 and the transmembrane and cytoplasmic domains of murine PIR-A3 showed the ability of PIR-A3 to physically interact with the FcɛRIγ chain in 293T cells. This interaction is dependent on Arg632 within the PIR-A3 transmembrane domain. We also demonstrate PIR-A3 interaction with the endogenous FcɛRIγ of the ANA-1 macrophage cell line, again in an Arg632-dependent manner. Furthermore, we show that crosslinking of these chimeric receptors synergizes with IFN-γ in the production of nitric oxide. Our data are the first to show the potential of PIR-A3 to deliver activation signals to macrophages and establish its dependence on Arg632. These findings suggest that further study of the PIR-A receptors should be aggressively pursued toward a complete understanding of the intricate regulation of macrophage biology.

Functional Association of FcɛRIγ With Arginine632 of Paired Immunoglobulin-Like Receptor (PIR)-A3 in Murine Macrophages

Blood ◽  
1999 ◽  
Vol 94 (5) ◽  
pp. 1790-1796
Author(s):  
Lynn S. Taylor ◽  
Daniel W. McVicar
Keyword(s):  
Transmembrane Domain ◽  
Dependent Manner ◽  
Macrophage Cell Line ◽  
Inhibitory Pathway ◽  
Macrophage Cell ◽  
Complete Understanding ◽  
Chimeric Receptors ◽  
Extracellular Region ◽  
Activating Receptors ◽  
Ifn Γ

Paired immunoglobulin-like receptors (PIR) are expressed on B cells and macrophages and include inhibitory and putative activating receptors referred to as PIR-B and PIR-A, respectively. Although PIR-B’s inhibitory pathway has been described, it is unknown whether PIR-A receptors can deliver activation signals to macrophages, and if so, through what mechanism. Here we use chimeric receptors to address the mechanisms of PIR-A signaling. Cotransfection of chimeric receptors comprised of the extracellular region of human CD4 and the transmembrane and cytoplasmic domains of murine PIR-A3 showed the ability of PIR-A3 to physically interact with the FcɛRIγ chain in 293T cells. This interaction is dependent on Arg632 within the PIR-A3 transmembrane domain. We also demonstrate PIR-A3 interaction with the endogenous FcɛRIγ of the ANA-1 macrophage cell line, again in an Arg632-dependent manner. Furthermore, we show that crosslinking of these chimeric receptors synergizes with IFN-γ in the production of nitric oxide. Our data are the first to show the potential of PIR-A3 to deliver activation signals to macrophages and establish its dependence on Arg632. These findings suggest that further study of the PIR-A receptors should be aggressively pursued toward a complete understanding of the intricate regulation of macrophage biology.

IFN-γ up-regulates kappa opioid receptors (KOR) on murine macrophage cell line J774

2012 ◽  
Vol 245 (1-2) ◽  
pp. 56-65 ◽  
Author(s):  
Jelka Gabrilovac ◽  
Barbara Čupić ◽  
Emilija Zapletal ◽  
Anamaria Brozovic
Keyword(s):  
Cell Line ◽  
Opioid Receptors ◽  
Murine Macrophage ◽  
Kappa Opioid Receptors ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Kappa Opioid ◽  
Murine Macrophage Cell Line ◽  
Ifn Γ

scholarly journals Interaction of Bartonella henselae with the Murine Macrophage Cell Line J774: Infection and Proinflammatory Response

2001 ◽  
Vol 69 (10) ◽  
pp. 5974-5980 ◽  
Author(s):  
Tiziana Musso ◽  
Raffaele Badolato ◽  
Daniela Ravarino ◽  
Sarah Stornello ◽  
Patrizia Panzanelli ◽  
...  
Keyword(s):  
Cell Line ◽  
Bartonella Henselae ◽  
Competitive Inhibitor ◽  
No Production ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Proinflammatory Response ◽  
Necrosis Factor Alpha ◽  
Ifn Γ ◽  
Limiting Condition

ABSTRACT Bartonella henselae is the causative agent of cat scratch disease (CSD), a self-limiting condition characterized by a subacute regional lymphadenopathy that may develop into disseminated bartonellosis in immunocompromised subjects. Mice experimentally infected with B. henselaedisplay typical liver and spleen granulomas rich in T cells and macrophages. So far there are no data on the interaction between bartonellae and macrophages. In order to clarify this topic, we investigated the interaction of B. henselae with J774, a mouse macrophage cell line. Analysis of bacterial uptake by functional assays and transmission electron microscopy indicates that bartonellae can enter and survive inside J774. Entry occurred within 30 min postinfection and reached a plateau at 160 min. Infection of J774 was followed by a dose-dependent release of the proinflammatory cytokines tumor necrosis factor alpha, interleukin 1β (IL-1β), and IL-6. Bartonellae persisted intracellularly without loss of viability for at least 8 h, and their number slightly decreased 24 h postinfection. Gamma interferon (IFN-γ) treatment of J774 significantly decreased the number of recoverable bacteria at 8 and 24 h. This enhancement of macrophage bactericidal activity was associated with nitric oxide (NO) release and was prevented by the addition of the competitive inhibitor of NO synthesis NG -monomethyll-arginine. These findings suggest that IFN-γ-mediated activation of macrophages may be important for the clearing ofB. henselae infection and that anti-B. henselae microbicidal activity of IFN-γ-activated macrophages is mediated to a large extent by NO production.

Zinc modulates mRNA levels of cytokines

2003 ◽  
Vol 285 (5) ◽  
pp. E1095-E1102 ◽  
Author(s):  
Bin Bao ◽  
Ananda S. Prasad ◽  
Frances W. J. Beck ◽  
Michele Godmere
Keyword(s):  
Gene Expression ◽  
Cell Line ◽  
Zinc Deficiency ◽  
Mrna Levels ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Th1 Cell ◽  
Killer Activity ◽  
Tnf Α ◽  
Ifn Γ

Zinc plays an important role in cell-mediated immune function. Altered cellular immune response resulting from zinc deficiency leads to frequent microbial infections, thymic atrophy, decreased natural killer activity, decreased thymic hormone activity, and altered cytokine production. In this study, we examined the effect of zinc deficiency on IL-2 and IFN-γ in HUT-78 (Th0) and D1.1 (Th1) cell lines and TNF-α, IL-1β, and IL-8 in the HL-60 (monocyte-macrophage) cell line. The results demonstrate that zinc deficiency decreased the levels of IL-2 and IFN-γ cytokines and mRNAs in HUT-78 after 6 h of PMA/ p-phytohemagglutinin (PHA) stimulation and in D1.1 cells after 6 h of PHA/ionomycin stimulation compared with the zinc-sufficient cells. However, zinc deficiency increased the levels of TNF-α, IL-1β, and IL-8 cytokines and mRNAs in HL-60 cells after 6 h of PMA stimulation compared with zinc-sufficient cells. Actinomycin D study suggests that the changes in the levels of these cytokine mRNAs were not the result of the stability affected by zinc but might be the result of altered expression of these cytokine genes. These data demonstrate that zinc mediates positively the gene expression of IL-2 and IFN-γ in the Th1 cell line and negatively TNF-α, IL-1β, and IL-8 in the monocyte-macrophage cell line. Our study shows that the effect of zinc on gene expression and production of cytokines is cell lineage specific.

scholarly journals A Tumor Necrosis Factor Mimetic Peptide Activates a Murine Macrophage Cell Line To Inhibit Mycobacterial Growth in a Nitric Oxide-Dependent Fashion

1998 ◽  
Vol 66 (5) ◽  
pp. 2122-2127 ◽  
Author(s):  
W. J. Britton ◽  
N. Meadows ◽  
D. A. Rathjen ◽  
D. R. Roach ◽  
H. Briscoe
Keyword(s):  
Nitric Oxide ◽  
Tumor Necrosis ◽  
Macrophage Activation ◽  
Tnf Receptor ◽  
Murine Macrophage ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Mycobacterial Infections ◽  
Ifn Γ ◽  
Necrosis Factor

ABSTRACT The control of mycobacterial infections depends on the cytokine-mediated activation of mononuclear phagocytes to inhibit the growth of intracellular mycobacteria. Optimal activation requires the presence of T-cell-derived gamma interferon (IFN-γ) and other signals, including tumor necrosis factor (TNF). Recently, an 11-mer peptide based on amino acids 70 to 80 of the human TNF sequence, TNF(70-80), was found to have TNF mimetic properties, which include the activation of human and mouse neutrophils to killPlasmodia spp. Therefore, we investigated the capacity of TNF(70-80) to activate the murine macrophage cell line RAW264.7 infected with the vaccine strain Mycobacterium bovis bacillus Calmette-Guérin (BCG). When RAW264.7 cells were pretreated with human TNF or TNF(70-80) in the presence of IFN-γ, there was a dose-dependent reduction in the replication of BCG as measured by the uptake of 3H-labeled uracil and a concomitant release of nitric oxide as measured by the nitrite in the culture supernatants. TNF- or TNF(70-80)-induced macrophage activation was dependent on IFN-γ and was inhibited by neutralizing monoclonal antibody to human TNF and by anti-IFN-γ antisera. Both nitrite release and BCG growth inhibition were abrogated by competitive inhibitors ofl-arginine, which blocked the activation of inducible nitric oxide synthase. A soluble form of the Type 1 TNF receptor blocked the activation of BCG-infected macrophages by human TNF and TNF(70-80), demonstrating that the effect of TNF(70-80) is dependent on signaling through TNF receptor I. The mimetic effects of TNF(70-80) on macrophage activation in vitro suggest that treatment with TNF(70-80) may modulate mycobacterial infections in vivo.

scholarly journals Exosomal arrow (Arr)/lipoprotein receptor protein 6 (LRP6) in Drosophila melanogaster increases the extracellular level of Sol narae (Sona) in a Wnt-independent manner

2020 ◽  
Vol 11 (11) ◽  
Author(s):  
Jeong-Hoon Han ◽  
Yeon Kim ◽  
Kyung-Ok Cho
Keyword(s):  
Cell Death ◽  
Transmembrane Domain ◽  
Premature Stop Codon ◽  
Receptor Protein ◽  
Dependent Manner ◽  
S2 Cells ◽  
Independent Manner ◽  
Extracellular Region ◽  
Extracellular Level ◽  
Intracellular Region

Abstract Wg/Wnt as a signaling protein binds to Frizzled (Fz) and Arrow (Arr), two Wg co-receptors essential for Wg signaling for cell proliferation, differentiation, and cell survival. Arr has a long extracellular region, a single transmembrane domain and an intracellular region. Here, we report that a new arrm7 mutant is identified in a genetic screen as a suppressor of lethality induced by overexpression of Sol narae (Sona), a secreted metalloprotease in ADAMTS family involved in Wg signaling. arrm7 allele has a premature stop codon, which encodes Arrm7 protein missing the intracellular region. arrm7 clones show cell death phenotype and overexpression of Arrm7 protein also induces cell death. Levels of extracellular Sona were decreased in both arrm7 and arr2 null clones, demonstrating that Arr increases the level of extracellular Sona. Indeed, Arr but not Arrm7, increased levels of Sona in cytoplasm and exosome fraction by inhibiting the lysosomal degradation pathway. Interestingly, Arr itself was identified in the exosome fraction, demonstrating that Arr is secreted to extracellular space. When Sona-expressing S2 cells were treated with exosomal Arr, the extracellular level of active Sona was increased. These results show that exosomal Arr dictates Sona-expressing cells to increase the level of extracellular Sona. This new function of Arr occurred in the absence of Wg because S2 cells do not express Wg. We propose that Arr plays two distinct roles, one as an exosomal protein to increase the level of extracellular Sona in a Wnt-independent manner and the other as a Wg co-receptor in a Wnt-dependent manner.

scholarly journals The Absence of PspA or Presence of Antibody to PspA Facilitates the Complement-Dependent Phagocytosis of PneumococciIn Vitro

2012 ◽  
Vol 19 (10) ◽  
pp. 1574-1582 ◽  
Author(s):  
Bing Ren ◽  
Jie Li ◽  
Kristopher Genschmer ◽  
Susan K. Hollingshead ◽  
David E. Briles
Keyword(s):  
Protein A ◽  
Surface Protein ◽  
Dependent Manner ◽  
Mouse Macrophage ◽  
Wild Type ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Type 3 ◽  
Complement Deposition ◽  
Mouse Macrophage Cell Line

ABSTRACTPneumococcal surface protein A (PspA) is a surface molecule on pneumococci that is required for full virulence in mouse models of infection. PspA has been reported to inhibit complement deposition on the pneumococcal surface. It has been assumed that this decreased complement deposition results in the inefficient phagocytosis of wild-type pneumococci. However, an effect of PspA on phagocytosis had not been shown. Our present studies demonstrated that a loss of PspA by capsular type 3 strains WU2 and A66.1 led to enhanced complement-dependent phagocytosis of the pneumococci by the mouse macrophage cell line J774A.1. This observation was made using human complement as well as mouse complement. Since this enhanced phagocytosis could be blocked by antibody to complement receptor CR3 on J774A.1, it was concluded that PspA's effect on phagocytosis was due to its effect on the amount of deposited complement, which in turn helped opsonize the pneumococci for phagocytosis. Since these studies included new independent mutants lacking PspA, the results provide solid confirmation of the previously reported effects of PspA on pneumococcal virulence and complement deposition. Finally, we showed that antibody to PspA, which is also known to enhance complement deposition, also enhances the phagocytosis of pneumococci in a largely complement-dependent manner.

scholarly journals In vitro Anti-inflammatory Effects of Satureja Kkhuzestanica Essential Oil Compared to Carvacrol

2020 ◽  
Vol 5 (2) ◽  
pp. 61-65
Author(s):  
Masumeh Jalalvand ◽  
Gholamreza Shahsavari ◽  
Ali Sheikhian ◽  
Ali Ganji ◽  
Ghasem Mosayebi
Keyword(s):  
Gene Expression ◽  
Essential Oil ◽  
Cell Line ◽  
Rna Extraction ◽  
Dependent Manner ◽  
Native Plant ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Mediators Of Inflammation ◽  
Anti Inflammatory

Introduction: Satureja khozestanica grows mainly in the southwest part of Iran as a native plant. This edible herb contains various compounds including the S. Khuzestanica essential oil (SKEO) and monoterpene known as Carvacrol. Previous studies have shown the anti-inflammatory effects of S. Khuzestanica without mentioning the exact mechanism of its function. Given that prostaglandin synthesis, which is one of the main mediators of inflammation, is regulated by the cyclooxygenase-2 (COX2) gene, the present study investigated the effects of SKEO and Carvacrol on the expression of the COX2 gene in the stimulated-J774A.1 macrophage cell line. Methods: To this end, fresh aerial parts of the plant were processed to prepare SKEO. Then, different doses of SKEO and Carvacrol (i.e., 0.004%, 0.008%, and 0.016% v/v) were used to treat with the lipopolysaccharides (LPS)-stimulated cell line for eight hours. After RNA extraction, the real-time polymerase chain reaction technique was applied for gene expression analysis. Results: In the LPS-stimulated J774A.1 macrophage cell line, COX2 gene expression reduced significantly in a dose-dependent manner (0.004%, 0.008%, and 0.016%, P = 0.024, P = 0.021, and P = 0.013 v/v of SKEO, respectively) by SKEO, and the effect of Carvacrol was less powerful (0.008% and 0.016%, P = 0.027 and P = 0.038 v/v, respectively) compared to SKEO. Finally, the comparison between SKEO and Carvacrol showed higher significant inhibitory effects of SKEO on COX2 gene expression in comparison with Carvacrol in 0.004% v/v concentration (P = 0.037). Conclusion: In general, SKEO significantly reduced COX2 gene expression, thus it can be suggested that its anti-inflammatory effect may result from the inhibition of the synthesis of this pro-inflammatory gene.

scholarly journals Carvacrol inhibits osteoclast differentiation induced by RANKL in RAW264.7 cells

2020 ◽  
Vol 11 (SPL4) ◽  
pp. 1779-1783
Author(s):  
Fathima Bushra Sheriff M ◽  
Amuthavalli Kottaiswamy ◽  
Deepa Suruli ◽  
Shila Samuel ◽  
Vijayaraghavan Radhakrishnan
Keyword(s):  
Osteoclast Differentiation ◽  
Cathepsin K ◽  
Raw264.7 Cells ◽  
Marker Genes ◽  
Dependent Manner ◽  
Macrophage Cell Line ◽  
Macrophage Cell ◽  
Trap Staining ◽  
Significant Change ◽  
Dose Dependent

RAW264.7, a murine macrophage cell line, an osteoclast model used for the differentiation of Osteoclast by inducing RANKL. Carvacrol, a monoterpenoid phenol that possess various medicinal property, was used for the treatment of osteoclast cells in this study. We investigate the effect of carvacrol in osteoclast cells induced by RANKL in RAW264.7 cells. RAW264.7 cells, cultured along with 40ng/ml of RANKL, on day 5, the cells were treated with TRAP staining to assess the formation of osteoclast cells, the presence of multinucleated TRAP positive cells were visualised using an inverted light microscope. The osteoclast cells were treated with varying concentrations of carvacrol (0-200µm) respectively for 48 hours. By MTT assay, it was found that there was no cytotoxic effect induced by carvacrol in RAW264.7 cells, whereas in RANKL induced osteoclast cells, there was a significant change in a dose dependent manner for 48 hours. By western blot and agarose gel electrophoresis, the levels of osteoclastogenic marker genes such as TRAP and cathepsin k were assessed and the developed osteoclast cells were treated with 150µm of carvacrol and found a significant change on treatment with carvacrol when compared with control. The present study reveals the anti-osteoclastogenic effect of carvacrol on osteoclast cells induced by RANKL in RAW264.7 cells.

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