Measurement of Macrophage Growth and Differentiation

Chiţu, Violeta; Yeung, Yee Guide; Yu, Wenfeng; Nandi, Sayan; Stanley, E. Richard

doi:10.1002/0471142735.im1420s92

Cited by 26 publications

(33 citation statements)

References 77 publications

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“…After Fc receptor blocking, cells were labeled with anti-CD11b-APC-Cy7, anti-CD115-PE, anti-Ly6C Pacific Blue, anti-Ly6G-PerCP, anti-F4/80-Pacific Blue, anti-B220-PE, anti-CD8a-PE-Cy5.5, anti-CD19-APC, anti-MHC II-fluorescein isothiocyanate, anti-CD206-APC, anti-IL-4Ra-PE, and anti-CD11c-fluorescein isothiocyanate (eBioscience). Macrophages 43,44 and peritoneal cells 45 were analyzed as described and sorted using a BD FACSAria II cell sorter (Becton Dickinson). Spectral overlaps between fluorophores were corrected by electronic compensation.…”

Section: -29mentioning

confidence: 99%

Colony stimulating factor-1 receptor signaling networks inhibit mouse macrophage inflammatory responses by induction of microRNA-21

Caescu

Guo

Tesfa³

et al. 2015

Blood

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134

118

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Key Points• Analysis of CSF-1R pTyrregulated messenger RNAs identifies novel signaling nodes and networks that can be targeted to modulate macrophage functions.• miR-21 is a novel CSF-1R pTyr-721-induced molecule that suppresses the macrophage M1 phenotype and enhances the M2 phenotype.Macrophage polarization between the M2 (repair, protumorigenic) and M1 (inflammatory) phenotypes is seen as a continuum of states. The detailed transcriptional events and signals downstream of colony-stimulating factor 1 receptor (CSF-1R) that contributes to amplification of the M2 phenotype and suppression of the M1 phenotype are largely unknown. Macrophage CSF-1R pTyr-721 signaling promotes cell motility and enhancement of tumor cell invasion in vitro. Combining analysis of cellular systems for CSF-1R gain of function and loss of function with bioinformatic analysis of the macrophage CSF-1R pTyr-721-regulated transcriptome, we uncovered microRNA-21 (miR-21) as a downstream molecular switch controlling macrophage activation and identified extracellular signal-regulated kinase 1/2 and nuclear factor-kB as CSF-1R pTyr-721-regulated signaling nodes. We show that CSF-1R pTyr-721 signaling suppresses the inflammatory phenotype, predominantly by induction of miR-21. Profiling of the miR-21-regulated messenger RNAs revealed that 80% of the CSF-1-regulated canonical miR-21 targets are proinflammatory molecules. Additionally, miR-21 positively regulates M2 marker expression. Moreover, miR-21 feeds back to positively regulate its own expression and to limit CSF-1R-mediated activation of extracellular signal-regulated kinase 1/2 and nuclear factor-kB. Consistent with an anti-inflammatory role of miRNA-21, intraperitoneal injection of mice with a miRNA-21 inhibitor increases the recruitment of inflammatory monocytes and enhances the peritoneal monocyte/macrophage response to lipopolysaccharide. These results identify the CSF-1R-regulated miR-21 network that modulates macrophage polarization. (Blood. 2015;125(8):e1-e13) IntroductionMacrophages protect the host against infection and injury and facilitate tissue remodeling.1 However, they frequently accumulate in pathological settings, including cancers, 2 atherosclerosis, 3 metabolic disease, 4 and sepsis, 5 where they respond to microenvironmental cues that can be detrimental to the host. Two distinct extreme states of polarized activation have been described in macrophages:6,7 the classically activated (M1) and the alternatively activated (M2) macrophage phenotypes, each characterized by well-described markers. 5,6,[8][9][10][11] M1 macrophages produce proinflammatory cytokines, elevate the expression of inducible nitric oxide synthase 2 (iNOS) and major histocompatibility complex class II (MHC II), 12 and can play antitumorigenic roles. 5,9 In contrast, the M2 macrophages have increased expression of scavenger receptors, increased activation of the arginase pathway, low expression of interleukin-12 (IL-12), high expression of IL-10 and IL-1RA, and increased anti-inflammatory responses a...

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Section: -29mentioning

confidence: 99%

Colony stimulating factor-1 receptor signaling networks inhibit mouse macrophage inflammatory responses by induction of microRNA-21

Caescu

Guo

Tesfa³

et al. 2015

Blood

Self Cite

134

118

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“…After red blood cell lysis and Fc receptor blocking, cells were labeled with anti-CD117-FITC, anti-CD11b-APC, biotinylated Rat IgG 2a anti-CSF-1R AFS98, and streptavidin-PE. Bone marrow populations containing myeloid and osteoclast precursors were identified as described, 8,20 and analyzed or sorted using a BD-LSR II, or a BDFACS Aria I cell sorter (Becton Dickinson), respectively. Spectral overlaps between fluorophores corrected by electronic compensation.…”

mentioning

confidence: 99%

PSTPIP2 deficiency in mice causes osteopenia and increased differentiation of multipotent myeloid precursors into osteoclasts

Chiţu

Nacu

Charles

et al. 2012

Blood

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Missense mutations that reduce or abrogate myeloid cell expression of the F-BAR domain protein, proline serine threonine phosphatase-interacting protein 2 (PST-PIP2), lead to autoinflammatory disease involving extramedullary hematopoiesis, skin and bone lesions. However, little is known about how PSTPIP2 regulates osteoclast development. Here we examined how PSTPIP2 deficiency causes osteopenia and bone lesions, using the mouse PSTPIP2 mutations, cmo, which fails to express PSTPIP2 and Lupo, in which PSTPIP2 is dysfunctional. In both models, serum levels of the pro-osteoclastogenic factor, MIP-1␣, were elevated and CSF-1 receptor (CSF-1R)-dependent production of MIP-1␣ by macrophages was increased. Treatment of cmo mice with a dual specificity CSF-1R and c-Kit inhibitor, PLX3397, decreased circulating MIP-1␣ and ameliorated the extramedullary hematopoiesis, inflammation, and osteopenia, demonstrating that aberrant myelopoiesis drives disease. Purified osteoclast precursors from PSTPIP2-deficient mice exhibit increased osteoclastogenesis in vitro and were used to probe the structural requirements for PST-PIP2 suppression of osteoclast development. PSTPIP2 tyrosine phosphorylation and a functional F-BAR domain were essential for PSTPIP2 inhibition of TRAP expression and osteoclast precursor fusion, whereas interaction with PEST-type phosphatases was only required for suppression of TRAP expression. Thus, PST-PIP2 acts as a negative feedback regulator of CSF-1R signaling to suppress inflammation and osteoclastogenesis. IntroductionThe combination of chronic immune activation and musculoskeletal tissue damage is the hallmark of rheumatic diseases. 1 Osteolytic lesions coupled with skin and/or joint inflammation occur in several rheumatic conditions, such as rheumatoid arthritis, psoriatic arthritis, and chronic recurrent multifocal osteomyelitis (CRMO). 1,2 Thus, an understanding of the pathophysiologic mechanisms underlying rheumatic disease requires the identification of the molecular pathways that simultaneously regulate inflammation and bone homeostasis.Osteoclasts are bone-resorbing multinucleated giant cells of myeloid origin. Receptor activator of nuclear factor B ligand (RANKL) and colony stimulating factor-1 (CSF-1) are necessary and sufficient for osteoclast differentiation from monocytic precursor cells in vivo and in vitro. 3-5 CSF-1 modulates multiple steps of osteoclastogenesis, including proliferation of mononuclear OC precursors (OCP), their differentiation and their fusion. In synergy with RANKL, CSF-1 also stimulates the expression of several osteoclast-specific genes including RANK, components of RANK signaling pathways and tartrate-resistant acid phosphatase (TRAP). [6][7][8][9] Proline serine threonine phosphatase-interacting protein 2 (PST-PIP2), also known as macrophage F-actin-associated and tyrosine phosphorylated protein (MAYP), is a Fes CIP4 homology domain (FCH) and Bin/Amphiphysin/Rvs (BAR; F-BAR) protein, predominantly expressed in the myeloid lineage. 10 It is rapidly tyrosine phosph...

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“…The ability of rocaglates to synergize with low concentrations of IFNγ, inhibit the prepared in our laboratory is critically dependent on CSF1 for survival (45). Nevertheless, increasing the CSF1-containing conditioned media 3-fold did not improve cell survival in the presence of rocaglates.…”

Section: Macrophage Activation By Rocaglates Improves Bacterial Controlmentioning

confidence: 72%

Channeling macrophage polarization via selective translation inhibition by rocaglates increases macrophage resistance to Mycobacterium tuberculosis

Chatterjee

Yabaji

Bhattacharya

et al. 2019

Preprint

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24The increasing problem of infectious diseases caused by antibiotic-resistant bacteria has 25 intensified a search for novel host-directed therapies aimed at either boosting immune-mediated 26 bacterial control or reducing immunopathology. Macrophages play central roles in eliminating 27 pathogens as well as in tissue homeostasis and repair via alternative activation programs, M1-28 like and M2-like, respectively. Macrophages of both phenotypes are often present at the sites of 29 chronic unresolved inflammation, such infectious granulomas and solid tumors, where the M2-30 like macrophages generate microenvironment conducive for pathogen survival or tumor 31 progression. 32Interferon-gamma is a major cytokine that drives M1-like macrophage activation, which 33 is essential for control of intracellular bacterial pathogens. To evaluate M1-like macrophage 34 polarization as a broad therapeutic strategy, we searched for small molecules capable of 35 enhancing effects of low concentrations of IFNγ on M1 macrophage polarization. We 36 identified synthetic rocaglates that potentiated macrophages responses to low concentrations 37 of IFNγ. Although known as potent translation inhibitors in tumor cell lines, in primary 38 macrophages select rocaglates induced the upregulation of IRF1, a master regulator of IFNγ 39-activated pathways, stimulated autophagy and increased macrophage resilience to oxidative 40 stress. In contrast, rocaglates inhibited macrophage responsiveness to IL-4, a prototypical 41 activator of the M2-like phenotype. The M1-like macrophage programing by rocaglates was 42 mechanistically linked to selective translation inhibition and modulation of MAP kinase 43 network. 44Thus, rocaglates represent a novel class of immunomodulators that can enforce 45 macrophage polarization towards the M1-like phenotype in complex microenvironments 46 3 associated with hypofunction of type 1 and/or hyperactivation of type 2 immunity, e.g., 47chronic bacterial infections, allergies and, possibly, certain tumors. 48 49 50 51The increasing problem of infectious diseases caused by antibiotic-resistant bacteria has 52 intensified a search for novel therapeutic approaches. Among them are host-directed therapies 53 aimed at either boosting immune-mediated bacterial control or reducing immunopathology, 54 broadly referred to as mechanisms of host resistance or disease tolerance, respectively (1). 55Many intracellular bacterial pathogens reside in macrophages, the very cells of innate immune 56 system whose major function is to eliminate invading pathogens. This paradox is enabled by 57 macrophages' plasticity, which successful pathogens exploit to create cellular niches for 58 persistence and replication within inflamed tissue of susceptible hosts. 59To combat intracellular bacteria, macrophages activate cell autonomous defense 60 mechanisms, such as phagocytosis, production of highly toxic reactive oxygen and nitrogen 61 species, bactericidal peptides, as well as phagosome maturation and autophagy to deliver the 62 ingested pat...

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Measurement of Macrophage Growth and Differentiation

Cited by 26 publications

References 77 publications

Colony stimulating factor-1 receptor signaling networks inhibit mouse macrophage inflammatory responses by induction of microRNA-21

Colony stimulating factor-1 receptor signaling networks inhibit mouse macrophage inflammatory responses by induction of microRNA-21

PSTPIP2 deficiency in mice causes osteopenia and increased differentiation of multipotent myeloid precursors into osteoclasts

Channeling macrophage polarization via selective translation inhibition by rocaglates increases macrophage resistance to Mycobacterium tuberculosis

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