Human Monocyte-Derived Macrophages (MDM): Model 1 (GM-CSF)

2022 ◽  
pp. 91-96
Author(s):  
Claudia Alteri ◽  
Lorenzo Piermatteo ◽  
Francesca Ceccherini Silberstein ◽  
Valentina Svicher ◽  
Carlo Federico Perno
Keyword(s):  
Human Monocyte ◽  
Gm Csf

scholarly journals Gamma-interferon modulates human monocyte/macrophage transferrin receptor expression

Blood ◽  
1988 ◽  
Vol 71 (6) ◽  
pp. 1590-1595 ◽  
Author(s):  
R Taetle ◽  
JM Honeysett
Keyword(s):  
Binding Sites ◽  
Blood Cells ◽  
Interleukin 1 ◽  
Human Monocyte ◽  
Gamma Interferon ◽  
Receptor Expression ◽  
Biologically Active ◽  
Gm Csf ◽  
Fe Content ◽  
Slot Blot Analysis

Although circulating human monocytes do not express transferrin (Tf) receptors, cultured adherent blood cells display high-affinity Tf binding sites. In the present studies, effects of various cytokines and biologically active proteins on human monocyte/macrophage Tf receptors were investigated. After culture, Tf receptor expression by adherent blood cells was time dependent and plateaued by 7 days. The addition of interleukin-1 (IL-1), alpha-interferon (alpha-IFN), granulocyte/macrophage-colony stimulating factor (GM-CSF), or human IgG to macrophages cultured for 4 days did not alter Tf receptor expression. Fe-saturated, human Tf caused a significant, dose-dependent decrease in receptor expression. At a dose of 100 U/mL, gamma- interferon (gamma-IFN) significantly increased Tf receptor expression by macrophages cultured for 4 (230% +/- 51% of control) or 7 days (150% +/- 22%). Scatchard analyses showed increased binding sites but no change in receptor affinity. Northern and slot blot analysis of cellular mRNA from macrophages cultured for 4 to 7 days and exposed to gamma-IFN showed a two- to fivefold increase in Tf receptor mRNA, but less than or equal to 30% increase in beta-actin mRNA. Ferritin content of gamma-IFN-treated macrophages was 47% to 63% of control cells. Net uptake of 59Fe from Tf by gamma-IFN-treated cells was 10% to 17% of control, but uptake of radiolabeled Tf was comparable. When macrophages were labeled with 59Fe and then exposed to gamma-IFN, cell-associated Fe was reduced by 43%, indicating that gamma-IFN caused macrophage Fe release. gamma-IFN specifically modulates Tf receptor display by inducing Fe release and reducing cellular Fe content. Regulation of Tf receptor expression in macrophages is controlled by cellular Fe content and is thus similar to regulatory mechanisms in dividing cells.

scholarly journals Recombinant human granulocyte/macrophage colony-stimulating factor activates intracellular killing of Leishmania donovani by human monocyte-derived macrophages.

1987 ◽  
Vol 166 (5) ◽  
pp. 1436-1446 ◽  
Author(s):  
W Y Weiser ◽  
A Van Niel ◽  
S C Clark ◽  
J R David ◽  
H G Remold
Keyword(s):  
Leishmania Donovani ◽  
Human Monocyte ◽  
Peripheral Blood Monocyte ◽  
Antileishmanial Activity ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Recombinant granulocyte/macrophage colony-stimulating factor (rGM-CSF) obtained from cloned complementary Mo cell DNA and expressed in COS-1 cells activates cultured peripheral blood monocyte-derived macrophages in vitro to become cytotoxic for intracellular L. donovani. The antileishmanial effect of rGM-CSF, which can be completely neutralized by anti-rGM-CSF antiserum, is maximal after 36 h preincubation with the cultured macrophages, compared with that of rIFN-gamma, which reaches its maximum at 72 h of preincubation. The antileishmanial effect of GM-CSF as well as IFN-gamma is independent of detectable amounts of LPS and is not augmented by the addition of 10 or 50 ng/ml of LPS. Simultaneous administration of suboptimal doses of rGM-CSF and rIFN-gamma to monocyte-derived macrophages results in greater antileishmanial activity by these cells than administration of either lymphokine alone, although no enhancement of antileishmanial activity is observed when optimal doses of these two lymphokines are applied together.

scholarly journals Gamma-interferon modulates human monocyte/macrophage transferrin receptor expression

Blood ◽  
1988 ◽  
Vol 71 (6) ◽  
pp. 1590-1595 ◽  
Author(s):  
R Taetle ◽  
JM Honeysett
Keyword(s):  
Binding Sites ◽  
Blood Cells ◽  
Interleukin 1 ◽  
Human Monocyte ◽  
Gamma Interferon ◽  
Receptor Expression ◽  
Biologically Active ◽  
Gm Csf ◽  
Fe Content ◽  
Slot Blot Analysis

Abstract Although circulating human monocytes do not express transferrin (Tf) receptors, cultured adherent blood cells display high-affinity Tf binding sites. In the present studies, effects of various cytokines and biologically active proteins on human monocyte/macrophage Tf receptors were investigated. After culture, Tf receptor expression by adherent blood cells was time dependent and plateaued by 7 days. The addition of interleukin-1 (IL-1), alpha-interferon (alpha-IFN), granulocyte/macrophage-colony stimulating factor (GM-CSF), or human IgG to macrophages cultured for 4 days did not alter Tf receptor expression. Fe-saturated, human Tf caused a significant, dose-dependent decrease in receptor expression. At a dose of 100 U/mL, gamma- interferon (gamma-IFN) significantly increased Tf receptor expression by macrophages cultured for 4 (230% +/- 51% of control) or 7 days (150% +/- 22%). Scatchard analyses showed increased binding sites but no change in receptor affinity. Northern and slot blot analysis of cellular mRNA from macrophages cultured for 4 to 7 days and exposed to gamma-IFN showed a two- to fivefold increase in Tf receptor mRNA, but less than or equal to 30% increase in beta-actin mRNA. Ferritin content of gamma-IFN-treated macrophages was 47% to 63% of control cells. Net uptake of 59Fe from Tf by gamma-IFN-treated cells was 10% to 17% of control, but uptake of radiolabeled Tf was comparable. When macrophages were labeled with 59Fe and then exposed to gamma-IFN, cell-associated Fe was reduced by 43%, indicating that gamma-IFN caused macrophage Fe release. gamma-IFN specifically modulates Tf receptor display by inducing Fe release and reducing cellular Fe content. Regulation of Tf receptor expression in macrophages is controlled by cellular Fe content and is thus similar to regulatory mechanisms in dividing cells.

scholarly journals The Meningococcal Adhesin NhhA Provokes Proinflammatory Responses in Macrophages via Toll-Like Receptor 4-Dependent and -Independent Pathways

2012 ◽  
Vol 80 (11) ◽  
pp. 4027-4033 ◽  
Author(s):  
Mikael Sjölinder ◽  
Georg Altenbacher ◽  
Xiao Wang ◽  
Yumin Gao ◽  
Charlotta Hansson ◽  
...  
Keyword(s):  
Gene Regulation ◽  
Gene Activation ◽  
Adaptor Protein ◽  
Human Monocyte ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Proinflammatory Responses ◽  
Macrophage Colony ◽  
Related Proteins ◽  
Stimulating Factor

ABSTRACTActivation of macrophages by Toll-like receptors (TLRs) and functionally related proteins is essential for host defense and innate immunity. TLRs recognize a wide variety of pathogen-associated molecules. Here, we demonstrate that the meningococcal outer membrane protein NhhA has immunostimulatory functions and triggers release of proinflammatory cytokines from macrophages. NhhA-induced cytokine release was found to proceed via two distinct pathways in RAW 264.7 macrophages. Interleukin-6 (IL-6) secretion was dependent on activation of TLR4 and required the TLR signaling adaptor protein MyD88. In contrast, release of tumor necrosis factor (TNF) was TLR4 and MyD88 independent. Both pathways involved NF-κB-dependent gene regulation. Using a PCR-based screen, we could identify additional targets of NhhA-dependent gene activation such as the cytokines and growth factors IL-1α, IL-1β, granulocyte colony-stimulating factor (G-CSF), and granulocyte-macrophage colony-stimulating factor (GM-CSF). In human monocyte-derived macrophages, G-CSF, GM-CSF, and IL-6 were found to be major targets of NhhA-dependent gene regulation. NhhA induced transcription of IL-6 and G-CSF mRNA via TLR4-dependent pathways, whereas GM-CSF transcription was induced via TLR4-independent pathways. These data provide new insights into the role of NhhA in host-pathogen interaction.

scholarly journals Macrophage colony-stimulating factor and granulocyte-macrophage colony- stimulating factor stimulate the synthesis of plasminogen-activator inhibitors by human monocytes

Blood ◽  
1993 ◽  
Vol 82 (12) ◽  
pp. 3616-3621 ◽  
Author(s):  
JA Hamilton ◽  
GA Whitty ◽  
H Stanton ◽  
J Wojta ◽  
M Gallichio ◽  
...  
Keyword(s):  
Plasminogen Activator ◽  
Human Monocyte ◽  
Mrna Levels ◽  
Human Monocytes ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor ◽  
Pai 1

Macrophage colony-stimulating factor (M-CSF or CSF-1) and granulocyte- macrophage CSF (GM-CSF) have been shown to increase human monocyte urokinase-type plasminogen-activator (u-PA) activity with possible consequences for cell migration and tissue remodeling; because monocyte u-PA activity is likely to be controlled in part also by the PA inhibitors (PAIs) made by the cell, the effect of M-CSF and GM-CSF on human monocyte PAI-2 and PAI-1 synthesis was investigated. To this end, elutriation-purified human monocytes were treated in vitro with purified recombinant human M-CSF and GM-CSF, and PAI-2 and PAI-1 antigen and mRNA levels measured by specific enzyme-linked immunosorbent assays and Northern blot, respectively. Each CSF could enhance the protein and mRNA levels of PAI-2 and PAI-1 at similar concentrations for each product. This similar regulation of monocyte PAI expression in response to the CSFs contrasted with that found for the effects of lipopolysaccharide, transforming growth factor-beta and a glucocorticoid. Therefore, PAIs may be modulating the effects of the CSFs on monocyte u-PA activity at sites of inflammation and tissue remodeling.

Immunosuppressive Effect of Cordyceps CS-4 on Human Monocyte-Derived Dendritic Cellsin Vitro

2010 ◽  
Vol 38 (05) ◽  
pp. 961-972 ◽  
Author(s):  
Jing Tang ◽  
Dan Tian ◽  
Gaolin Liu
Keyword(s):  
T Cells ◽  
B Lymphocytes ◽  
Cd4 T Cells ◽  
Fruit Body ◽  
Human Monocyte ◽  
Immunosuppressive Effect ◽  
Th2 Response ◽  
Gm Csf ◽  
Antigen Presenting ◽  
Th1 And Th2

Cordyceps CS-4 (C.CS-4), a vegetative form of Cordyceps that contains the same active compounds as the fruit body, is widely used as a substitute of Cordyceps in China. A number of studies have shown that Cordyceps can positively stimulate the activation of T lymphocytes, B lymphocytes, natural killer cells, and macrophages. In our previous study, we found that C.CS-4 could inhibit the proliferation of CD4+ T cells in autoimmune diseases and prevent the lymphocyte infiltration in tissues. However, it is still unclear how the lymphocytes are regulated by C.CS-4. In this study, we investigate the effect of C.CS-4 on human monocyte-derived dendritic cells ( Mo -DCs), which are generated from PBMCs by the treatment with GM-CSF and IL-4. It is observed that Mo -DCs pretreated with C.CS-4 show an immature phenotype. Moreover, C.CS-4 significantly inhibits proliferation of CD4+ T cells, attenuates the production of cytokines in Mo -DCs and balances the Th1 and Th2 response in immune system. Our findings indicate that C.CS-4 exerts the immunosuppressive effect through inhibiting the CD4+ T cells proliferation, regulating cytokine secretions of Th1 and Th2 response ( Mo -DCs) and inducing phenotypic immature of Mo -DCs which may be related to the antigen presenting dysfunction.

scholarly journals Monoclonal Antibodies Capable of Binding SARS-CoV-2 Spike Protein Receptor Binding Motif Specifically Prevent GM-CSF Induction

2020 ◽  
Author(s):  
Xiaoling Qiang ◽  
Shu Zhu ◽  
Jianhua Li ◽  
Ping Wang ◽  
Kevin J. Tracey ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Receptor Binding ◽  
Viral Entry ◽  
Protein Targeting ◽  
Inflammatory Responses ◽  
Human Monocyte ◽  
Spike Protein ◽  
Binding Motif ◽  
Gm Csf ◽  
Future Assessment

AbstractA severe acute respiratory syndrome (SARS)-like coronavirus (SARS-CoV-2) has recently caused a pandemic COVID-19 disease that infected more than 25.6 million and killed 852,000 people worldwide. Like the SARS-CoV, SARS-CoV-2 also employs a receptor-binding motif (RBM) of its envelope spike protein for binding the host angiotensin-converting enzyme 2 (ACE2) to gain viral entry. Currently, extensive efforts are being made to produce vaccines against a surface fragment of a SARS-CoV-2, such as the spike protein, in order to boost protective antibody responses. It was previously unknown how spike protein-targeting antibodies would affect innate inflammatory responses to SARS-CoV-2 infections. Here we generated a highly purified recombinant protein corresponding to the RBM of SARS-CoV-2, and used it to screen for cross-reactive monoclonal antibodies (mAbs). We found two RBM-binding mAbs that competitively inhibited its interaction with human ACE2, and specifically blocked the RBM-induced GM-CSF secretion in both human monocyte and murine macrophage cultures. Our findings have suggested a possible strategy to prevent SARS-CoV-2-elicited “cytokine storm”, and provided a potentially useful criteria for future assessment of innate immune-modulating properties of various SARS-CoV-2 vaccines.One Sentence SummaryRBM-binding Antibodies Inhibit GM-CSF Induction.

scholarly journals Regulation of human monocyte DNA synthesis by colony-stimulating factors, cytokines, and cyclic adenosine monophosphate

Blood ◽  
1992 ◽  
Vol 79 (8) ◽  
pp. 1972-1981 ◽  
Author(s):  
DL Cheung ◽  
JA Hamilton
Keyword(s):  
Dna Synthesis ◽  
Human Peripheral Blood ◽  
Cyclic Adenosine Monophosphate ◽  
Human Monocyte ◽  
Adenosine Monophosphate ◽  
Interleukin 4 ◽  
Murine Macrophage ◽  
Colony Stimulating Factor ◽  
Gm Csf ◽  
Stimulating Factor

Abstract It is reported in this study that a subpopulation of highly purified human peripheral blood human monocytes can proliferate in response to colony-stimulating factor-1 (CSF-1), granulocyte-macrophage colony- stimulating factor (GM-CSF), and interleukin-3 (IL-3). Both GM-CSF and IL-3 synergized with CSF-1 for the induction of DNA synthesis. Given the DNA synthesis levels attained, we were able to test the effects of certain cytokines and cyclic adenosine monophosphate (cAMP)-elevating agents, which have been shown to modulate in vitro human myelopoiesis and murine macrophage proliferation. The cytokines, interferon-gamma (IFN-gamma), interleukin-4 (IL-4), and tumor necrosis factor-alpha (TNF- alpha), as well as cAMP-elevating agents, 8-bromoadenosine 3′:5′-cyclic monophosphate (8BrcAMP), cholera toxin (CT), and prostaglandin E2 (PGE2), suppressed the monocyte DNA synthesis due to CSF-1. These results parallel those reported with human bone marrow progenitors, as well as murine macrophage populations. The cycling human monocyte population could provide a model cell type to understand the molecular events controlling human myelopoiesis.

Primary Acute Myeloid Leukemic Blasts Display Constitutive Serine 585 Phosphorylation within the 14-3-3 Binding Motif of the GM-CSF/IL-3 Receptor Required for Survival Signalling.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 70-70 ◽  
Author(s):  
Daniel Thomas ◽  
Jason Powell ◽  
Szu-Hee Lee ◽  
Angel F. Lopez ◽  
Guthridge Mark
Keyword(s):  
Cell Survival ◽  
Adaptor Protein ◽  
Human Monocyte ◽  
Buffy Coat ◽  
Binding Motif ◽  
Beta Chain ◽  
Blood Monocytes ◽  
Gm Csf ◽  
Threonine Kinases ◽  
Acute Myeloid

Abstract Acute myeloid leukemia (AML) is characterised by defects in the regulation of hematopoietic cell survival, differentiation and proliferation. IL-3 and GM-CSF exert potent survival and proliferative effects on a range of myeloid progenitor cells. Previously we have identified a novel cytoplasmic motif of the human GM-CSF/IL-3 receptor beta chain responsible for coordinating survival and proliferative responses in primary hematopoietic cells. The motif contains a serine residue (585) that is phosphorylated in response to cytokine and couples to the PI3K/AKT pathway via the 14-3-3 adaptor protein and is required for cell survival (Guthridge et al, Molecular Cell6:99, 2000;Blood103:820, 2004). An adjacent tyrosine residue (577) within the motif is phosphorylated via Jak2 and associates with proliferative function. We investigated the phosphorylation status of the beta chain motif in response to GM-CSF stimulation in blasts cells from 12 patients with AML, and in blood monocytes from 6 healthy volunteers. Leukemic samples consisted of >99% blasts after buffy coat separation according to cytospin morphology. All leukemic blasts and blood monocyte samples expressed alpha and beta chain by flow cytometry. The phosphorylation status of residues 577 and 585 were examined utilising beta chain phospho-specific antibodies, after stimulation of AML blasts or blood monocytes with GM-CSF and beta chain immunoprecipitation. Human monocyte preparations all showed inducible serine 585 phosphorylation and tyrosine 577 phosphorylation. In contrast AML blasts displayed strong constitutive serine 585 phosphorylation in 10 out of 12 patients. No discernible tyrosine 577 phosphorylation in the absence of cytokine was observed for the AML samples. Beta chain phosphorylation pattern was independent of FAB classification. Inhibition of Jak2 did not prevent serine 585 phosphorylation of the receptor. Constitutive Akt activation was observed in AML blasts. The data provides evidence for dysregulation of a novel GM-CSF/IL-3 receptor mediated survival pathway involving serine/threonine kinases, 14-3-3 and Akt that is active in primary AML blasts in the majority of patients. This data would suggest that dysregulated activation of serine/threonine kinases and/or phosphatases at the receptor level may contribute to leukemogenesis.

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