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1.
Adv Exp Med Biol ; 1185: 181-186, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31884609

RESUMO

As the resident macrophages of central nervous system, microglia reside in the plexiform and nerve fiber layers of the retina. In degenerative diseases, monocyte-derived macrophages can be recruited to the retina, and histopathology shows abnormal accumulation of macrophages subretinally. However, due to lack of known markers, recruited cells and resident microglia are phenotypically indistinguishable, leaving a major knowledge gap about their potentially independent roles. Here, we used single cell RNA-seq and analyzed over 10,000 immune cells of mouse retinas from normal control and light damage-induced retinal degeneration. We observed ten major macrophage clusters. Moreover, combining trajectory analysis and in situ validation allowed us to pinpoint that subretinal phagocytes are microglia-derived and express high levels of Gal3, Cd68, and Lpl but not P2ry12. Hence, we have identified novel subretinal macrophage markers indicative of their origin and phenotype, which may be useful in other degeneration models and human specimens.


Assuntos
Microglia/classificação , Degeneração Retiniana/patologia , Animais , Modelos Animais de Doenças , Humanos , Macrófagos/classificação , Macrófagos/citologia , Camundongos , Microglia/citologia , RNA-Seq , Retina/citologia
2.
Nat Commun ; 10(1): 4353, 2019 09 25.
Artigo em Inglês | MEDLINE | ID: mdl-31554795

RESUMO

Stat6 is known to drive macrophage M2 polarization. However, how macrophage polarization is fine-tuned by Stat6 is poorly understood. Here, we find that Lys383 of Stat6 is acetylated by the acetyltransferase CREB-binding protein (CBP) during macrophage activation to suppress macrophage M2 polarization. Mechanistically, Trim24, a CBP-associated E3 ligase, promotes Stat6 acetylation by catalyzing CBP ubiquitination at Lys119 to facilitate the recruitment of CBP to Stat6. Loss of Trim24 inhibits Stat6 acetylation and thus promotes M2 polarization in both mouse and human macrophages, potentially compromising antitumor immune responses. By contrast, Stat6 mediates the suppression of TRIM24 expression in M2 macrophages to contribute to the induction of an immunosuppressive tumor niche. Taken together, our findings establish Stat6 acetylation as an essential negative regulatory mechanism that curtails macrophage M2 polarization.


Assuntos
Ativação de Macrófagos , Macrófagos/metabolismo , Neoplasias Experimentais/metabolismo , Proteínas Nucleares/metabolismo , Fator de Transcrição STAT6/metabolismo , Fatores de Transcrição/metabolismo , Acetilação , Animais , Linhagem Celular Tumoral , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/genética , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Regulação da Expressão Gênica , Células HEK293 , Humanos , Lisina/genética , Lisina/metabolismo , Macrófagos/classificação , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Neoplasias Experimentais/genética , Neoplasias Experimentais/patologia , Proteínas Nucleares/genética , Fator de Transcrição STAT6/genética , Fatores de Transcrição/genética
3.
BMC Immunol ; 20(1): 34, 2019 09 18.
Artigo em Inglês | MEDLINE | ID: mdl-31533615

RESUMO

BACKGROUND: Resident macrophages (Mø) originating from yolk sac Mø and/or foetal monocytes colonise tissues/organs during embryonic development. They persist into adulthood by self-renewal at a steady state, independent of adult monocyte inputs, except for those in the intestines and dermis. Thus, many resident Mø can be propagated in vitro under optimal conditions; however, there are no specific in vitro culture methods available for the propagation of resident Mø from diverse tissues/organs. RESULTS: We provided a simple method for propagating resident Mø derived from the liver, spleen, lung, and brain of ICR male mice by co-culture and subculture along with the propagation of other stromal cells of the respective organs in standard culture media and successfully demonstrated the propagation of resident Mø colonising these organs. We also proposed a simple method for segregating Mø from stromal cells according to their adhesive property on bacteriological Petri dishes, which enabled the collection of more than 97.6% of the resident Mø from each organ. Expression analyses of conventional Mø markers by flow cytometry showed similar expression patterns among the Mø collected from the organs. CONCLUSION: This is the first study to clearly provide a practical Mø propagation method applicable to resident Mø of diverse tissues and organs. Thus, this novel practical Mø propagation method can offer broad applications for the use of resident Mø of diverse tissues and organs.


Assuntos
Técnicas de Cultura de Células , Macrófagos/citologia , Macrófagos/metabolismo , Animais , Biomarcadores , Adesão Celular , Técnicas de Cocultura , Citocinas/metabolismo , Citofagocitose/imunologia , Imunofenotipagem , Ativação de Macrófagos , Macrófagos/classificação , Camundongos , Especificidade de Órgãos
4.
Biochemistry (Mosc) ; 84(7): 729-745, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31509725

RESUMO

Despite the progress of modern medicine, oncological diseases are still among the most common causes of death of adult populations in developed countries. The current therapeutic approaches are imperfect, and the high mortality of oncological patients under treatment, the lack of personalized strategies, and severe side effects arising as a result of treatment force seeking new approaches to therapy of malignant tumors. During the last decade, cancer immunotherapy, an approach that relies on activation of the host antitumor immune response, has been actively developing. Cancer immunotherapy is the most promising trend in contemporary fundamental and practical oncology, and restoration of the pathologically altered tumor microenvironment is one of its key tasks, in particular, the reprogramming of tumor macrophages from the immunosuppressive M2-phenotype into the proinflammatory M1-phenotype is pivotal for eliciting antitumor response. This review describes the current knowledge about macrophage classification, mechanisms of their polarization, their role in formation of the tumor microenvironment, and strategies for changing the functional activity of M2-macrophages, as well as problems of targeted delivery of immunostimulatory signals to tumor macrophages using nanoparticles.


Assuntos
Imunoterapia , Macrófagos/metabolismo , Nanopartículas/metabolismo , Neoplasias/terapia , Animais , Polaridade Celular/fisiologia , Humanos , Fatores Imunológicos/farmacologia , Fatores Imunológicos/uso terapêutico , Microscopia Intravital , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos/química , Macrófagos/classificação , Camundongos , Nanopartículas/química , Fenótipo , Coroa de Proteína/imunologia , Microambiente Tumoral/imunologia
5.
Nature ; 572(7771): 670-675, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31391580

RESUMO

Macrophages are considered to contribute to chronic inflammatory diseases such as rheumatoid arthritis1. However, both the exact origin and the role of macrophages in inflammatory joint disease remain unclear. Here we use fate-mapping approaches in conjunction with three-dimensional light-sheet fluorescence microscopy and single-cell RNA sequencing to perform a comprehensive spatiotemporal analysis of the composition, origin and differentiation of subsets of macrophages within healthy and inflamed joints, and study the roles of these macrophages during arthritis. We find that dynamic membrane-like structures, consisting of a distinct population of CX3CR1+ tissue-resident macrophages, form an internal immunological barrier at the synovial lining and physically seclude the joint. These barrier-forming macrophages display features that are otherwise typical of epithelial cells, and maintain their numbers through a pool of locally proliferating CX3CR1- mononuclear cells that are embedded into the synovial tissue. Unlike recruited monocyte-derived macrophages, which actively contribute to joint inflammation, these epithelial-like CX3CR1+ lining macrophages restrict the inflammatory reaction by providing a tight-junction-mediated shield for intra-articular structures. Our data reveal an unexpected functional diversification among synovial macrophages and have important implications for the general role of macrophages in health and disease.


Assuntos
Articulações/citologia , Macrófagos/citologia , Macrófagos/fisiologia , Membrana Sinovial/citologia , Sinoviócitos/citologia , Sinoviócitos/fisiologia , Junções Íntimas/fisiologia , Animais , Artrite/imunologia , Artrite/patologia , Receptor 1 de Quimiocina CX3C/análise , Receptor 1 de Quimiocina CX3C/metabolismo , Rastreamento de Células , Feminino , Perfilação da Expressão Gênica , Humanos , Inflamação/imunologia , Inflamação/patologia , Articulações/patologia , Macrófagos/classificação , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Análise de Componente Principal , Análise de Célula Única , Sinoviócitos/classificação , Sinoviócitos/metabolismo , Transcriptoma/genética
6.
Res Vet Sci ; 125: 382-389, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31404885

RESUMO

Toxoplasma gondii is an intracellular opportunistic, parasitic protozoan. Microglia have been classified into two main types: M1 (classically activated macrophages) and M2 (alternatively activated macrophages). BV2 cells were used in this study, together with lipopolysaccharide (LPS) and interferon (IFN)-γ or interleukin (IL)-4, which were used to induce resting microglia. Expression levels of M1/M2 markers were determined at both mRNA and protein levels, using PCR, western blot, and flow cytometry. Furthermore, cells were infected with T. gondii PLK strain, and the dynamic changes in M1/M2 marker expression levels were determined. An in vitro polarization model was successfully established. Expression of Nos2 and M1-associated markers was significantly upregulated at 12 h post-infection in BV2 cells. Further, the JAK/STAT1 and NF-κB signaling pathways were also activated following T. gondii infection. This demonstrated that T. gondii infection induces M1-type microglial polarization in vitro. The present study demonstrated that T. gondii infection affects microglial activation in vitro and elucidated the effects of activated microglia on T. gondii proliferation. This data may serve as a useful reference for more detailed elucidation of interactions between T. gondii and the innate immune system.


Assuntos
Macrófagos/parasitologia , Microglia/parasitologia , Toxoplasma/fisiologia , Animais , Linhagem Celular , Proliferação de Células , Regulação da Expressão Gênica , Interferon gama/farmacologia , Interleucina-4/farmacologia , Lipopolissacarídeos/farmacologia , Macrófagos/classificação , NF-kappa B/metabolismo , RNA Mensageiro/metabolismo , Transdução de Sinais
7.
Hum Immunol ; 80(10): 890-896, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31447056

RESUMO

The functional phenotype of macrophages (Mφ) is determined by both differentiation factors and polarization stimuli. In mouse Mφ could be easily divided into the distinct Mφ subtypes. However, the identification of human M1 and M2 cells is much more difficult due to the lack of M1- or M2-specific markers. We assumed that the Mφ capacity to induce T cell proliferation in mixed leukocyte culture, or allostimulatory activity, may be a marker of Mφ functional phenotype. We compared the allostimulatory activity of Mφ differentiated with GM-CSF or M-CSF and polarized into M1, M2a, M2c subtypes using appropriate stimuli. GM-CSF-differentiated M1 Mφ showed pronounced allostimulatory activity whereas the polarization into M2a and M2c of GM-CSF-differentiated Mφ was associated with decreased allostimulatory activity. M-CSF-differentiated M1 Mφ demonstrated the moderate increasing of allostimulatory activity but its level has never reached that of GM-CSF-activated M1. The level of allostimulatory activity of M2a and M2c M-CSF-induced Mφ was comparable to that of GM-CSF-induced M2a and M2c Mφ. Thus, low allostimulatory activity is a common property of human M2a and M2c macrophages regardless of the differentiating factor and a polarizing stimulus and can be used to distinguish between M1 and M2 phenotypes.


Assuntos
Polaridade Celular/efeitos dos fármacos , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Ativação de Macrófagos/efeitos dos fármacos , Fator Estimulador de Colônias de Macrófagos/farmacologia , Macrófagos/imunologia , Fenótipo , Adulto , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/imunologia , Proliferação de Células , Células Cultivadas , Dexametasona/farmacologia , Feminino , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Voluntários Saudáveis , Humanos , Interleucina-4/farmacologia , Lipopolissacarídeos/farmacologia , Fator Estimulador de Colônias de Macrófagos/metabolismo , Macrófagos/classificação , Masculino , Pessoa de Meia-Idade , Curva ROC , Proteínas Recombinantes , Adulto Jovem
8.
PLoS Comput Biol ; 15(7): e1007172, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31365522

RESUMO

In an inflammatory setting, macrophages can be polarized to an inflammatory M1 phenotype or to an anti-inflammatory M2 phenotype, as well as existing on a spectrum between these two extremes. Dysfunction of this phenotypic switch can result in a population imbalance that leads to chronic wounds or disease due to unresolved inflammation. Therapeutic interventions that target macrophages have therefore been proposed and implemented in diseases that feature chronic inflammation such as diabetes mellitus and atherosclerosis. We have developed a model for the sequential influx of immune cells in the peritoneal cavity in response to a bacterial stimulus that includes macrophage polarization, with the simplifying assumption that macrophages can be classified as M1 or M2. With this model, we were able to reproduce the expected timing of sequential influx of immune cells and mediators in a general inflammatory setting. We then fit this model to in vivo experimental data obtained from a mouse peritonitis model of inflammation, which is widely used to evaluate endogenous processes in response to an inflammatory stimulus. Model robustness is explored with local structural and practical identifiability of the proposed model a posteriori. Additionally, we perform sensitivity analysis that identifies the population of apoptotic neutrophils as a key driver of the inflammatory process. Finally, we simulate a selection of proposed therapies including points of intervention in the case of delayed neutrophil apoptosis, which our model predicts will result in a sustained inflammatory response. Our model can therefore provide hypothesis testing for therapeutic interventions that target macrophage phenotype and predict outcomes to be validated by subsequent experimentation.


Assuntos
Inflamação/imunologia , Macrófagos/imunologia , Modelos Imunológicos , Animais , Apoptose/imunologia , Biologia Computacional , Simulação por Computador , Modelos Animais de Doenças , Humanos , Mediadores da Inflamação/imunologia , Ativação de Macrófagos , Macrófagos/classificação , Macrófagos Peritoneais/classificação , Macrófagos Peritoneais/imunologia , Camundongos , Neutrófilos/citologia , Neutrófilos/imunologia , Fenótipo
9.
Immunohorizons ; 3(8): 412-421, 2019 08 27.
Artigo em Inglês | MEDLINE | ID: mdl-31455692

RESUMO

Infection with the intestinal parasite Giardia duodenalis is one of the most common causes of diarrheal disease in the world. Previous work has demonstrated that the cells and mechanisms of the adaptive immune system are critical for clearance of this parasite. However, the innate system has not been as well studied in the context of Giardia infection. We have previously demonstrated that Giardia infection leads to the accumulation of a population of CD11b+, F4/80+, ARG1+, and NOS2+ macrophages in the small intestinal lamina propria. In this report, we sought to identify the accumulation mechanism of duodenal macrophages during Giardia infection and to determine if these cells were essential to the induction of protective Giardia immunity. We show that F4/80+, CD11b+, CD11cint, CX3CR1+, MHC class II+, Ly6C-, ARG1+, and NOS2+ macrophages accumulate in the small intestine during infections in mice. Consistent with this resident macrophage phenotype, macrophage accumulation does not require CCR2, and the macrophages incorporate EdU, indicating in situ proliferation rather than the recruitment of monocytes. Depletion of macrophages using anti-CSF1R did not impact parasite clearance nor development of regulatory T cell or Th17 cellular responses, suggesting that these macrophages are dispensable for protective Giardia immunity.


Assuntos
Giardia lamblia/imunologia , Giardíase/imunologia , Macrófagos/imunologia , Animais , Proliferação de Células/efeitos dos fármacos , Citocinas/genética , Nucleotídeos de Desoxiuracil/administração & dosagem , Nucleotídeos de Desoxiuracil/farmacologia , Duodeno/imunologia , Duodeno/parasitologia , Técnicas de Inativação de Genes , Giardíase/parasitologia , Intestino Delgado/imunologia , Macrófagos/classificação , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Membrana Mucosa/imunologia , Fenótipo , Receptores CCR2/genética , Receptores CCR2/metabolismo , Células Th17/imunologia
10.
Immunohorizons ; 3(7): 262-273, 2019 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-31356156

RESUMO

Bone marrow (BM)-derived classical monocytes are critical to wound repair, where they differentiate into macrophages and purge foreign materials and dead cells while also laying the framework for tissue repair and regeneration. A subset of this recruited population persists in the wound and acquires alternative activation states to promote cell proliferation and matrix remodeling. In diabetes, this phenotypic switch is impaired and inflammation persists in an elevated state, contributing to delayed wound healing. Long-term tissue-resident macrophages can also play a key role in the resolution of inflammation to varying degrees across different organs. In this study, we investigated different macrophage subpopulations in nondiabetic and diabetic wounds over time using Cx3CR1eGFP transgenic mice and BM transplants. We show Cx3CR1eGFP-hi macrophages in skin wounds are derived from long-term tissue-resident macrophages and predominantly exhibit an alternative activation state, whereas cells expressing low-intermediate Cx3CR1eGFP are derived from the BM, contribute to both early and later stages of wound healing, and show both classical and alternative activation states. Diabetic mice showed significant differences in the dynamics of these subpopulations, which likely contribute to elevated and persisting inflammatory states over time. In particular, failure of Cx3CR1int macrophages to mature into Cx3CR1hi links maturation to resolution of inflammation. Thus strategies to promote macrophage maturation may be effective therapeutic tools in chronic inflammatory environments.


Assuntos
Receptor 1 de Quimiocina CX3C/metabolismo , Diabetes Mellitus Experimental/metabolismo , Macrófagos/metabolismo , Cicatrização/fisiologia , Animais , Células da Medula Óssea , Transplante de Medula Óssea , Receptor 1 de Quimiocina CX3C/genética , Diferenciação Celular , Proliferação de Células , Modelos Animais de Doenças , Proteínas de Fluorescência Verde/metabolismo , Inflamação/metabolismo , Macrófagos/classificação , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos NOD , Camundongos Transgênicos , Monócitos/metabolismo , Receptores CCR2/metabolismo , Receptores para Leptina/deficiência
11.
Front Biosci (Landmark Ed) ; 24: 1271-1283, 2019 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-31136979

RESUMO

Macrophages are essential elements in the tumor microenvironment, where they can promote tumor growth but also influence the efficacy of anticancer strategies. In conventional therapies, chemotherapy and radiotherapy, TAMs play a dichotomous role, contributing to antitumor activity or hindering the efficacy of cytoreductive therapies. Macrophages express checkpoint ligands and are therefore targets of immunotherapy approaches based on checkpoint inhibitors. Targeted therapies with monoclonal antibodies elicit TAMs to engage in antitumor functions such as antibody-dependent phagocytosis through the activation of Fc receptors. New approaches to exploit macrophage effector functions induced by therapeutic antibodies are under investigation. Finally, strategies aimed at targeting TAM recruitment, survival and functional polarization are advancing towards the clinic. Collectively, TAM-centered strategies will hopefully complement conventional and unconventional anticancer therapies to achieve improved therapeutic benefit.


Assuntos
Anticorpos Monoclonais Humanizados/uso terapêutico , Imunoterapia/métodos , Macrófagos/imunologia , Neoplasias/terapia , Microambiente Tumoral/efeitos dos fármacos , Animais , Anticorpos Monoclonais Humanizados/imunologia , Humanos , Macrófagos/classificação , Macrófagos/metabolismo , Modelos Imunológicos , Neoplasias/imunologia , Neoplasias/metabolismo , Microambiente Tumoral/imunologia
12.
Dev Comp Immunol ; 98: 20-33, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-30974109

RESUMO

We previously demonstrated that the most bioactive vitamin A metabolite, all-trans retinoic acid (ATRA), increased T helper 2-associated responses induced in pigs by infection with the parasitic nematode Ascaris suum We also showed that ATRA potentiated the mRNA expression of several IL-4 induced chemokines (chemokine (CC motif) ligand 11 [(CCL11), CCL17, CCL22 and CCL26] associated with alternative activation (M2a) in porcine macrophages in vitro. Herein, several mechanisms whereby ATRA affects IL-4 signaling are profiled using large-scale real time PCR and RNA-Seq analysis. Twenty-three genes associated with M2a markers in other species were independently upregulated by both IL-4 and ATRA, including the adenosine receptor A2B (ADORA2B), cysteinyl leukotriene receptor 2 (CYSLTR2) and the vitamin D receptor (VDR). ATRA synergistically enhanced IL-4 up-regulation of Hepatitis A virus cellular receptor 2 (HAVCR2) and transglutaminase 2 (TGM2) and further repressed IL-4 down-regulated CD163 and Cytochrome b-245, beta polypeptide (CYBB) mRNA. Macrophages treated with ATRA exhibited a dose-dependent reduction in phagocytosis of opsonized Staphylococcus aureus. In addition, the combination of IL-4 and ATRA up-regulated the anti-inflammatory protein, IL-1R antagonist (IL1RN) and TGM2. These data indicate that ATRA induces a state of partial alternative activation in porcine macrophages, and amplifies certain aspects of M2a activation induced by IL-4. Given the prevalence of allergic and parasitic diseases worldwide and the close similarities in the porcine and human immune responses, these findings have important implications for the nutritional regulation of allergic inflammation at mucosal surfaces.


Assuntos
Ativação de Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Fagocitose/efeitos dos fármacos , Tretinoína/farmacologia , Animais , Células Cultivadas , Quimiocinas/genética , Quimiocinas/imunologia , Humanos , Interleucina-4/farmacologia , Ativação de Macrófagos/genética , Ativação de Macrófagos/imunologia , Macrófagos/classificação , Macrófagos/metabolismo , Fagocitose/imunologia , Staphylococcus aureus/imunologia , Suínos , Transcriptoma/efeitos dos fármacos , Transcriptoma/imunologia
13.
J Immunol Res ; 2019: 2368249, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30931335

RESUMO

Tumour-associated macrophage (TAM) serves as the site in which most inflammatory cells coreside. It plays an important role in determining the progression and metastasis of a tumour. The characteristic of TAM is largely dependent on the stimuli present in its tumour microenvironment (TME). Under this environment, however, M2 macrophages are found to be in abundance compared to M1 macrophages which later promote tumour progression. Numerous studies have elucidated the relationship between TAM and the progression of tumour; hence, TAM has now been the subject of interest among researchers for anticancer therapy. This review discusses the role of TAM in colorectal cancer (CRC) and some of the potential candidates that could reeducate TAM to fight against CRC. It is with hope that this review will serve as the foundation in understanding TAM in CRC and helping other researchers to select the most suitable candidate to reeducate TAM that could assist in enhancing the tumouricidal activity of M1 macrophage and eventually repress the development of CRC.


Assuntos
Neoplasias do Colo/imunologia , Neoplasias do Colo/terapia , Macrófagos/imunologia , Microambiente Tumoral/imunologia , Progressão da Doença , Humanos , Macrófagos/classificação , Fenótipo
14.
Cytokine Growth Factor Rev ; 46: 36-44, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30910350

RESUMO

Osteoarthritis (OA) is the most common form of arthritic disease, leading to disability and impaired quality of life and no curative treatments exist. Increasing evidence indicates that low-grade inflammation plays a pivotal role in the onset and progression of OA. In this review, we summarize emerging findings on the pathological roles of synovial macrophages, adipose tissue macrophages, and osteoclasts in OA and their potential clinical implications from cell biology to preclinical and preliminary clinical trials. The failure of synovial macrophages to transition from pro-inflammatory M1 to anti-inflammatory M2 subtypes may contribute to the initiation and maintenance of synovitis in OA. M1 macrophages promote the inflammatory microenvironment and progression of OA through interactions with synovial fibroblasts and chondrocytes, thus increasing the secretion of matrix metalloproteinases. Direct inhibition of M1 or promotion of M2 polarization may be useful therapeutic interventions. Adipose tissue macrophages present in the infrapatella fat pad (IPFP) were involved in the progression of obesity-induced OA, which contributed to changes in the integrity of the IPFP. Furthermore, macrophages and osteoclasts in the subchondral bone were involved in bone remodeling and pain through uncoupled osteoclast/osteoblast activity and increased nociceptive signaling. Growing evidence has indicated an important role for macrophage-mediated low-grade inflammation in OA. Fully understanding the link between macrophages and other cells in joints will provide new insights into OA disease modification.


Assuntos
Inflamação/fisiopatologia , Macrófagos/patologia , Osteoartrite do Joelho/imunologia , Osteoartrite do Joelho/fisiopatologia , Animais , Ensaios Clínicos como Assunto , Citocinas/imunologia , Humanos , Macrófagos/classificação , Camundongos , Osteoclastos/fisiologia , Qualidade de Vida
15.
Mar Drugs ; 17(2)2019 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-30717366

RESUMO

Macrophages are central mediators of inflammation, orchestrating the inflammatory response through the production of cytokines and nitric oxide. Macrophages obtain pro-inflammatory (M1) and anti-inflammatory (M2) phenotypes, which can be modulated by soluble factors, including natural products. Despite the crucial protective role of inflammation, chronic or deregulated inflammation can lead to pathological states, such as autoimmune diseases, metabolic disorders, cardiovascular diseases, and cancer. In this case, we studied the anti-inflammatory activity of neorogioltriol (1) in depth and identified two structurally related diterpenes, neorogioldiol (2), and O11,15-cyclo-14-bromo-14,15-dihydrorogiol-3,11-diol (3), with equally potent activity. We investigated the mechanism of action of metabolites 1⁻3 and found that all three suppressed macrophage activation and promoted an M2-like anti-inflammatory phenotype by inducing expression of Arginase1, MRC1, IRAK-M, the transcription factor C/EBPß, and the miRNA miR-146a. In addition, they suppressed iNOS induction and nitric oxide production. Importantly, treatment of mice with 2 or 3 suppressed DSS-induced colitis by reducing tissue damage and pro-inflammatory cytokine production. Thus, all these three diterpenes are promising lead molecules for the development of anti-inflammatory agents targeting macrophage polarization mechanisms.


Assuntos
Diterpenos/química , Diterpenos/farmacologia , Doenças Inflamatórias Intestinais/tratamento farmacológico , Laurencia/química , Macrófagos/efeitos dos fármacos , Animais , Proliferação de Células , Sulfato de Dextrana/toxicidade , Doenças Inflamatórias Intestinais/induzido quimicamente , Doenças Inflamatórias Intestinais/patologia , Macrófagos/classificação , Camundongos , Camundongos Endogâmicos C57BL , Estrutura Molecular , Células RAW 264.7
16.
Immunity ; 50(2): 418-431.e6, 2019 02 19.
Artigo em Inglês | MEDLINE | ID: mdl-30770245

RESUMO

Sepsis is a bi-phasic inflammatory disease that threatens approximately 30 million lives and claims over 14 million annually, yet little is known regarding the molecular switches and pathways that regulate this disease. Here, we have described ABCF1, an ATP-Binding Cassette (ABC) family member protein, which possesses an E2 ubiquitin enzyme activity, through which it controls the Lipopolysaccharide (LPS)- Toll-like Receptor-4 (TLR4) mediated gram-negative insult by targeting key proteins for K63-polyubiquitination. Ubiquitination by ABCF1 shifts the inflammatory profile from an early phase MyD88-dependent to a late phase TRIF-dependent signaling pathway, thereby regulating TLR4 endocytosis and modulating macrophage polarization from M1 to M2 phase. Physiologically, ABCF1 regulates the shift from the inflammatory phase of sepsis to the endotoxin tolerance phase, and modulates cytokine storm and interferon-ß (IFN-ß)-dependent production by the immunotherapeutic mediator, SIRT1. Consequently, ABCF1 controls sepsis induced mortality by repressing hypotension-induced renal circulatory dysfunction.


Assuntos
Transportadores de Cassetes de Ligação de ATP/imunologia , Macrófagos/imunologia , Sepse/imunologia , Choque Séptico/imunologia , Enzimas de Conjugação de Ubiquitina/imunologia , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/metabolismo , Trifosfato de Adenosina/imunologia , Trifosfato de Adenosina/metabolismo , Animais , Citocinas/imunologia , Citocinas/metabolismo , Feminino , Interferon beta/imunologia , Interferon beta/metabolismo , Lipopolissacarídeos/administração & dosagem , Lipopolissacarídeos/imunologia , Ativação de Macrófagos/efeitos dos fármacos , Ativação de Macrófagos/genética , Ativação de Macrófagos/imunologia , Macrófagos/classificação , Macrófagos/metabolismo , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Interferência de RNA , Sepse/genética , Sepse/metabolismo , Choque Séptico/genética , Choque Séptico/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Receptor 4 Toll-Like/imunologia , Receptor 4 Toll-Like/metabolismo , Enzimas de Conjugação de Ubiquitina/genética , Enzimas de Conjugação de Ubiquitina/metabolismo , Ubiquitinação/imunologia
17.
Cell Immunol ; 336: 75-82, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30665661

RESUMO

Obesity is seen as a low grade inflammatory state, and is associated with adverse pregnancy outcomes. Disturbed macrophage characteristics might be essential in obesity associated pregnancy pathology via effects on the regulation of angiogenesis and placental development. This study aims to address the effects of maternal obesity on macrophage subsets in the decidua of women with term uncomplicated pregnancies. Macrophages were isolated from the decidua basalis and decidua parietalis of women with pre-gravid BMI < 25 (control) and BMI > 30 (obese). Macrophages were characterized and quantified using multi-color flow cytometry. Placentas of 10 obese and 10 control women after an uncomplicated term pregnancy were included. The decidua parietalis, but not decidua basalis, showed significantly lower levels of M1-type (HLA-DR+, CD163-) macrophages (p < 0.05) in obese women (4,3% of total macrophages) compared to control women (5,3% of total macrophages). The lower levels of M1 macrophages, considered to be pro-inflammatory, might indicate a mechanism to compensate for the pro-inflammatory environment in obese women to ensure healthy pregnancy outcomes.


Assuntos
Decídua/imunologia , Macrófagos/classificação , /imunologia , Adulto , Antígenos CD/análise , Antígenos de Diferenciação Mielomonocítica/análise , Feminino , Antígenos HLA-DR/análise , Humanos , Gravidez , Receptores de Superfície Celular/análise
18.
Cancer Immunol Immunother ; 68(2): 189-200, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30341559

RESUMO

As a major component of the microenvironment of solid tumors, tumor-associated macrophages (TAMs) facilitate tumor progression. Intermediate-sized hyaluronan (INT-HA) fragments have an immunological function in cell differentiation; however, their role in promoting the polarization of non-activated macrophages to an M2-like TAM phenotype has not been characterized, and the underlying mechanisms remain unclear. Here, we used a miRNA microarray to find that some miRNAs (especially miR-935) were differentially regulated in INT-HA-induced M2-like macrophages. According to RT-qPCR and Western blot, there was an association between miR-935 and C/EBPß, that control the polarization of macrophages. Moreover, we found that INT-HA induced an M2-like phenotype via the TLR4 receptor. In our study, there was a negative correlation between plasma HA and miR-935 in monocytes from the peripheral blood of patients with solid tumors. There was also a negative correlation between miR-935 and M2-like macrophage markers in monocytes. These findings suggest that HA fragments interact with TLR4 and educate macrophage polarization to an M2-like phenotype via miR-935. Therefore, this study provides new insight into the role of miR-935 in INT-HA-induced M2-like polarization, and suggests a potential therapeutic target for antitumor treatment.


Assuntos
Diferenciação Celular/imunologia , Ácido Hialurônico/metabolismo , Macrófagos/imunologia , MicroRNAs/imunologia , Monócitos/imunologia , Receptor 4 Toll-Like/imunologia , Adulto , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Células Cultivadas , Feminino , Perfilação da Expressão Gênica , Humanos , Ácido Hialurônico/química , Ácido Hialurônico/farmacologia , Ativação de Macrófagos/efeitos dos fármacos , Ativação de Macrófagos/imunologia , Macrófagos/classificação , Macrófagos/metabolismo , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Peso Molecular , Monócitos/metabolismo , Neoplasias/genética , Neoplasias/imunologia , Neoplasias/metabolismo , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Células THP-1 , Receptor 4 Toll-Like/metabolismo
19.
Cancer Immunol Immunother ; 68(2): 269-282, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30430204

RESUMO

Aging immune deterioration and Epstein-Barr (EBV) intrinsic mechanisms play an essential role in EBV-positive diffuse large B-cell lymphoma (DLBCL) of the elderly (EBV + DLBCLe) pathogenesis, through the expression of viral proteins, interaction with host molecules and epigenetic regulation, such as miR-155, required for induction of M1 phenotype of macrophages. This study aims to evaluate the relationship between macrophage polarization pattern in the tumor microenvironment and relative expression of miR-155 in EBV + DLBCLe and EBV-negative DLBCL patients. We studied 28 EBV + DLBCLe and 65 EBV-negative DLBCL patients. Tumor-associated macrophages (TAM) were evaluated by expression of CD68, CD163 and CD163/CD68 ratio (degree of M2 polarization), using tissue microarray. RNA was extracted from paraffin-embedded tumor samples for miR-155 relative expression study. We found a significantly higher CD163/CD68 ratio in EBV + DLBCLe compared to EBV-negative DLBCL. In EBV-negative DLBCL, CD163/CD68 ratio was higher among advanced-staged/high-tumor burden disease and overexpression of miR-155 was associated with decreased polarization to the M2 phenotype of macrophages. The opposite was observed in EBV + DLBCLe patients: we found a positive association between miR-155 relative expression and CD163/CD68 ratio, which was not significant after outlier exclusion. We believe that the higher CD163/CD68 ratio in this group is probably due to the presence of the EBV since it directly affects macrophage polarization towards M2 phenotype through cytokine secretion in the tumor microenvironment. Therapeutic strategies modulating miR-155 expression or preventing immuno-regulatory and pro-tumor macrophage polarization could be adjuvants in EBV + DLBCLe therapy since this entity has a rich infiltration of M2 macrophages in its tumor microenvironment.


Assuntos
Infecções por Vírus Epstein-Barr/imunologia , Linfoma Difuso de Grandes Células B/imunologia , Macrófagos/imunologia , MicroRNAs/imunologia , Idoso , Idoso de 80 Anos ou mais , Antígenos CD/imunologia , Antígenos CD/metabolismo , Antígenos de Diferenciação Mielomonocítica/imunologia , Antígenos de Diferenciação Mielomonocítica/metabolismo , Infecções por Vírus Epstein-Barr/complicações , Infecções por Vírus Epstein-Barr/virologia , Feminino , Regulação Neoplásica da Expressão Gênica/imunologia , Herpesvirus Humano 4/imunologia , Herpesvirus Humano 4/fisiologia , Humanos , Linfoma Difuso de Grandes Células B/complicações , Linfoma Difuso de Grandes Células B/genética , Ativação de Macrófagos/imunologia , Macrófagos/classificação , Macrófagos/metabolismo , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Receptores de Superfície Celular/imunologia , Receptores de Superfície Celular/metabolismo , Microambiente Tumoral/genética , Microambiente Tumoral/imunologia
20.
São Paulo; s.n; s.n; 2019. 87 p. graf, tab.
Tese em Português | LILACS | ID: biblio-1015337

RESUMO

Os aminoácidos de cadeia ramificada (ACR) são considerados indispensáveis, pois não podem ser sintetizados endogenamente, sendo facilmente obtidos pela dieta. Entretanto, em determinadas condições clínicas, tanto a ingestão quando a absorção desses aminoácidos pode estar comprometida, levando ao estado hipercatabólico e prejudicando a função imune. O papel imunomodulador dos ACR tem sido relacionado com a melhora no balanço nitrogenado e o aumento da síntese e proliferação de células imunes, bem como, da síntese de mediadores inflamatórios. Entretanto, o mecanismo pelo qual os ACR exercem essas funções supracitadas ainda não é claro na literatura científica. Desta forma, esse trabalho teve como objetivo avaliar os efeitos da suplementação com ACR sobre os parâmetros inflamatórios e moleculares em macrófagos RAW 264.7 estimulados com lipopolissacarídeo (LPS). As culturas celulares foram distribuídas em cinco grupos: CTL - sem suplementação com ACR; LEU - suplementado com leucina (2 mmol/L); ISO - suplementado com isoleucina (2mmol/L); VAL - suplementado com valina (2 mmol/L) e LIV - suplementado com leucina (2 mmol/L), isoleucina (2 mmol/L) e valina (2 mmol/L). O estado inflamatório foi induzido pela adição de LPS (1 µg/mL) ao meio de cultura, seguindo quatro protocolos de tratamento: PT - pré-tratamento; TA - tratamento agudo; TC - tratamento crônico e TT - tratamento tardio. O ensaio de viabilidade celular foi realizado pelo teste MTT e a dosagem de óxido nítrico (NO) pela reação de Griess. As citocinas pró e anti-inflamatórias, e a prostaglandina E2 (PGE2) foram analisadas pelo método de ELISA. Para a avaliação dos parâmetros moleculares foi utilizado o método de western blotting. Houve aumento da viabilidade celular em todos os grupos suplementados em relação ao grupo controle no TA, no TC e no TT. Acerca da síntese de NO, a suplementação com ACR foi capaz de aumentar esse parâmetro em três dos quatro tratamentos propostos (PT, TA e TC). Em relação à síntese de citocinas pró e anti-inflamatórias, o PT e o TC foram mais eficazes em aumentar esse parâmetro em comparação aos outros tratamentos. Não houve diferença entre os grupos em relação à capacidade de síntese de PGE2 e à fosforilação de proteínas intracelulares. A partir dos resultados obtidos é possível concluir que os ACR contribuem significativamente para a viabilidade celular, bem como para a síntese de mediadores pró e anti-inflamatórios, sendo que o protocolo de suplementação se apresenta como fator determinante para obtenção desses resultados. Apesar da literatura científica atribuir grande parte dos efeitos imunomodulatórios à leucina, os resultados obtidos nesse estudo atribuem relevante potencial imunomodulador à isoleucina, abrindo espaço para um importante tema de estudo


Branched chain amino acids (BCAA) are considered indispensable, since they cannot be endogenously synthesized, being easily obtained by diet. However, in certain clinical conditions, both the intake and absorption of these amino acids may be compromised, leading to the hypercatabolic state and impairing the immune function. The immunomodulatory role of BCAA has been associated with the nitrogen balance improvement and the increase of production and proliferation of immune cells, as well as the synthesis of inflammatory mediators. However, the mechanisms by which BCAA modulate the immune system have not yet been completely elucidated. In this sense, this study aimed to evaluate the effects of BCAA supplementation on intracellular mechanisms and inflammatory parameters in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Cell cultures were distributed into five groups: CTL - without ACR supplementation; LEU - supplemented with leucine (2 mmol/L); ISO - supplemented with isoleucine (2mmol / L); VAL - supplemented with valine (2 mmol/L) and LIV - supplemented with leucine (2 mmol/L), isoleucine (2 mmol/L) and valine (2 mmol/L). The inflammatory state was induced by the addition of LPS (1 µg/ml) to the culture medium, following four treatment protocols: PT - pre-treatment; TA - acute treatment; TC - chronic treatment and TT - late treatment. The cell viability assay was performed by the MTT test and the nitric oxide (NO) dosage by the Griess reaction. Pro- and anti-inflammatory cytokines, and prostaglandin E2 (PGE2) were analyzed by ELISA. For the evaluation of the molecular parameters, the western blotting method was used. There was an increase in cell viability in all supplemented groups in relation to the control group in the TA, TC and TT treatments. Regarding NO synthesis, BCAA supplementation was able to increase NO production in three of the four proposed treatments (PT, TA and TC). In relation to the production of pro- and anti-inflammatory cytokines, PT and CT were more effective in increasing this parameter, compared to the other treatments. There was no difference between groups in relation to PGE2 production and intracellular protein phosphorylation. From the obtained results it is possible to conclude that the BCAA significantly contributed to the cell viability, as well as, for the production of pro and anti-inflammatory mediators, and the supplementation protocol presents as determinant factor to obtain these results. Although the scientific literature attributed a large part of the immunomodulatory effects to leucine, the results obtained in this study attribute relevant immunomodulatory potential to isoleucine, opening space for an important study topic


Assuntos
Animais , Masculino , Camundongos , Lipopolissacarídeos , Aminoácidos de Cadeia Ramificada/efeitos adversos , Inflamação/dietoterapia , Macrófagos/classificação
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