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1.
Fish Shellfish Immunol ; 94: 730-738, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31580934

RESUMO

Atrazine (ATR) causes environmental problems and damages the health of fish and aquatic animals. MicroRNAs (miRNAs) play important roles in immune regulation. However, the immunotoxicity mechanism of ATR in fish lymphocytes and the role of miRNA in this process remain unclear. To further study these mechanisms, spleen lymphocytes were exposed to 20, 40 and 60 µg/ml ATR for 18 h. Fluorescence staining and flow cytometry showed that the number of necrotic lymphocytes increased after ATR exposure. Compared with the control group, the mRNA expression of miR-181-5p was inhibited and the mRNA levels of TNF-α and HK2 were increased after ATR exposure. Additionally, the NF-κB inflammatory pathway and the levels of glycometabolism-related genes were upregulated. These results suggest that ATR induces inflammation and elevates glycometabolism in lymphocytes. We further found that the mRNA levels of receptor-interacting serine-threonine kinase 1 (RIP1), receptor-interacting serine-threonine kinase 3 (RIP3), mixed lineage kinase domain-like pseudokinase (MLKL), cylindromatosis (CYLD) and Fas-Associated protein with Death Domain (FADD) and the protein levels of RIP3 and MLKL in the treatment groups were significantly increased compared to those in control group, suggesting that ATR causes lymphocyte necroptosis. We conclude that miR-181-5p plays a key role in necroptosis in carp lymphocytes exposed to ATR by downregulating the expression of HK and TNF-α, which increases the level of glycometabolism and induces the inflammatory response, respectively.


Assuntos
Atrazina/toxicidade , Carpas/imunologia , Doenças dos Peixes/imunologia , Glicólise/imunologia , Inflamação/veterinária , MicroRNAs/metabolismo , /imunologia , Animais , Relação Dose-Resposta a Droga , Doenças dos Peixes/induzido quimicamente , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Glicólise/efeitos dos fármacos , Herbicidas/toxicidade , Inflamação/induzido quimicamente , Inflamação/imunologia , Linfócitos/efeitos dos fármacos , Linfócitos/imunologia , Baço/efeitos dos fármacos , Baço/imunologia
2.
Immunology ; 158(2): 104-120, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31318442

RESUMO

Activation of B and T lymphocytes leads to major remodelling of the metabolic landscape of the cells enabling their post-activation functions. However, naive B and T lymphocytes also show metabolic differences, and the genesis, nature and functional significance of these differences are not yet well understood. Here we show that resting B-cells appeared to have lower energy demands than resting T-cells as they consumed lower levels of glucose and fatty acids and produced less ATP. Resting B-cells are more dependent on OXPHOS, while T-cells show more dependence on aerobic glycolysis. However, despite an apparently higher energy demand, T lineage cells showed lower rates of protein synthesis than equivalent B lineage stages. These metabolic differences between the two lineages were established early during lineage differentiation, and were functionally significant. Higher levels of protein synthesis in B-cells were associated with increased synthesis of MHC class II molecules and other proteins associated with antigen internalization, transport and presentation. The combination of higher energy demand and lower protein synthesis in T-cells was consistent with their higher ATP-dependent motility. Our data provide an integrated perspective of the metabolic differences and their functional implications between the B and T lymphocyte lineages.


Assuntos
Linfócitos B/metabolismo , Glicólise/imunologia , Fosforilação Oxidativa , Linfócitos T/metabolismo , Trifosfato de Adenosina/biossíntese , Animais , Linfócitos B/citologia , Linfócitos B/imunologia , Diferenciação Celular/imunologia , Linhagem da Célula/genética , Linhagem da Célula/imunologia , Ácidos Graxos/metabolismo , Expressão Gênica , Glucose/metabolismo , Glicólise/genética , Antígenos de Histocompatibilidade Classe II/genética , Antígenos de Histocompatibilidade Classe II/imunologia , Imunofenotipagem , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Especificidade de Órgãos , Cultura Primária de Células , Biossíntese de Proteínas/imunologia , Linfócitos T/citologia , Linfócitos T/imunologia
3.
Med Oncol ; 36(9): 76, 2019 Jul 24.
Artigo em Inglês | MEDLINE | ID: mdl-31342270

RESUMO

The microenvironment of a tumor may regulate the anti-tumor immune response. Intratumoral acidosis and hypoxia may suppress lymphocyte proliferation and migration, and this may have important implications in modern immunotherapy. The expression of PD-L1 by cancer cells and of PD-1 by tumor infiltrating lymphocytes (TILs) was assessed in tissue specimens from 98 operable NSCLC patients. Their prognostic role and their association with makers of glycolysis and anaerobic metabolism were assessed. Strong cytoplasmic/membrane PD-L1 expression was noted in 45/98 cases. Intense presence of TILs was noted in 42/98 cases (high TIL-score), and intense presence of PD-1 expressing TILs (high PIL-score) in 17/98 cases. PD-L1 expression was directly correlated with high PIL-score (p = 0.005). A significant inverse relationship was found between lactate dehydrogenase LDH5 expression and PIL-score (p = 0.008). Similarly, low PIL-score was significantly linked with high-hexokinase HXKII and monocarboxylate transporter MCT2 expression (p < 0.04). Cases with both intense TIL-score and PIL-score had significantly better survival (p < 0.05). For patients with high TIL-score or high PIL-score, PD-L1 overexpression defined significantly poorer survival (p = 0.01 and p = 0.03, respectively). In multivariate analysis, stage (p = 0.002, HR 3.33, 95%CI 1.4-4.5) and TIL-score (p = 0.02, HR 2.12, 95%CI 1.1-4.0) were independent predictive variables of death events. Given the low specificity of PD-L1 as a biomarker for anti-PD-1/PD-L1 immunotherapy, a combined assessment of TIL, PD-L1, PD-1, and LDH5 provides a tool for an immunological/metabolic classification of NSCLC tumors, with a different prognosis and different expected response to anti-PD-1/PD-L1 immunotherapy, which should be considered in relevant clinical trials.


Assuntos
Antígeno B7-H1/genética , Carcinoma Pulmonar de Células não Pequenas/imunologia , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Glicólise/imunologia , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/metabolismo , Receptor de Morte Celular Programada 1/genética , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígeno B7-H1/metabolismo , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Carcinoma Pulmonar de Células não Pequenas/patologia , Feminino , Regulação Neoplásica da Expressão Gênica , Glicólise/genética , Hexoquinase/metabolismo , Humanos , Neoplasias Pulmonares/patologia , Linfócitos do Interstício Tumoral/imunologia , Linfócitos do Interstício Tumoral/patologia , Masculino , Pessoa de Meia-Idade , Transportadores de Ácidos Monocarboxílicos/metabolismo , Receptor de Morte Celular Programada 1/metabolismo , Análise de Sobrevida , Microambiente Tumoral
4.
Redox Biol ; 26: 101255, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31247505

RESUMO

Nearly 130 years after the first insights into the existence of mitochondria, new rolesassociated with these organelles continue to emerge. As essential hubs that dictate cell fate, mitochondria integrate cell physiology, signaling pathways and metabolism. Thus, recent research has focused on understanding how these multifaceted functions can be used to improve inflammatory responses and prevent cellular dysfunction. Here, we describe the role of mitochondria on the development and function of immune cells, highlighting metabolic aspects and pointing out some metabolic- independent features of mitochondria that sustain cell function.


Assuntos
Imunidade Adaptativa , Sistema Imunitário/fisiologia , Imunidade Inata , Mitocôndrias/imunologia , Dinâmica Mitocondrial/imunologia , /imunologia , Animais , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Glicólise/imunologia , Humanos , Inflamassomos/imunologia , Inflamassomos/metabolismo , Linfócitos/imunologia , Linfócitos/metabolismo , Macrófagos/imunologia , Macrófagos/metabolismo , Mitocôndrias/metabolismo , Neutrófilos/imunologia , Neutrófilos/metabolismo , Oxirredução , Fosforilação Oxidativa
5.
Nat Commun ; 10(1): 2042, 2019 05 03.
Artigo em Inglês | MEDLINE | ID: mdl-31053703

RESUMO

Metabolic pathways that regulate T-cell function show promise as therapeutic targets in diverse diseases. Here, we show that at rest cultured human effector memory and central memory CD4+ T-cells have elevated levels of glycolysis and oxidative phosphorylation (OXPHOS), in comparison to naïve T-cells. Despite having low resting metabolic rates, naive T-cells respond to TCR stimulation with robust and rapid increases in glycolysis and OXPHOS. This early metabolic switch requires Akt activity to support increased rates of glycolysis and STAT5 activity for amino acid biosynthesis and TCA cycle anaplerosis. Importantly, both STAT5 inhibition and disruption of TCA cycle anaplerosis are associated with reduced IL-2 production, demonstrating the functional importance of this early metabolic program. Our results define STAT5 as a key node in modulating the early metabolic program following activation in naive CD4+ T-cells and in turn provide greater understanding of how cellular metabolism shapes T-cell responses.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptores de Antígenos de Linfócitos T/metabolismo , Fator de Transcrição STAT5/metabolismo , Linfócitos T CD4-Positivos/metabolismo , Células Cultivadas , Ciclo do Ácido Cítrico/imunologia , Glicólise/imunologia , Voluntários Saudáveis , Humanos , Memória Imunológica , Ativação Linfocitária , Fosforilação Oxidativa , Cultura Primária de Células , Proteínas Proto-Oncogênicas c-akt/imunologia , Receptores de Antígenos de Linfócitos T/imunologia , Fator de Transcrição STAT5/imunologia
6.
Nat Commun ; 10(1): 1685, 2019 04 11.
Artigo em Inglês | MEDLINE | ID: mdl-30976008

RESUMO

Neonatal sepsis is characterized by hyperinflammation causing enhanced morbidity and mortality compared to adults. This suggests differences in the response towards invading threats. Here we investigate activated cord blood macrophages (CBMΦ) in comparison to adult macrophages (PBMΦ), indicating incomplete interferon gamma (IFN-γ) and interleukin 10 (IL-10)-induced activation of CBMΦ. CBMΦ show reduced expression of phagocytosis receptors and cytokine expression in addition to altered energy metabolism. In particular, IFN-γ as well as IL-10-activated CBMΦ completely fail to increase glycolysis and furthermore show reduced activation of the mTOR pathway, which is important for survival in sepsis. MTOR inhibition by rapamycin equalizes cytokine production in CBMΦ and PBMΦ. Finally, incubation of PBMΦ with cord blood serum or S100A8/A9, which is highly expressed in neonates, suppresses mTOR activation, prevents glycolysis and the expression of an PBMΦ phenotype. Thus, a metabolic alteration is apparent in CBMΦ, which might be dependent on S100A8/A9 expression.


Assuntos
Citotoxicidade Imunológica , Metabolismo Energético/imunologia , Macrófagos/metabolismo , Adulto , Fatores Etários , Calgranulina A/imunologia , Calgranulina A/metabolismo , Calgranulina B/imunologia , Calgranulina B/metabolismo , Diferenciação Celular/imunologia , Células Cultivadas , Sangue Fetal/citologia , Glicólise/imunologia , Voluntários Saudáveis , Humanos , Recém-Nascido , Interferon gama/imunologia , Interferon gama/metabolismo , Interleucina-10/imunologia , Interleucina-10/metabolismo , Macrófagos/imunologia , Cultura Primária de Células , Sepse/imunologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologia , Sirolimo/farmacologia , Serina-Treonina Quinases TOR/imunologia , Serina-Treonina Quinases TOR/metabolismo
7.
Monoclon Antib Immunodiagn Immunother ; 38(2): 60-69, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31009338

RESUMO

CD28 superagonist (CD28SA), a therapeutic immunomodulatory monoclonal antibody triggered rapid and exaggerated activation of CD4+ effector memory T cells (TEMs) in humans with unwanted serious adverse effects. It is well known that distinct metabolic programs determine the fate and responses of immune cells. In this study, we show that human CD4+ TEMs stimulated with CD28SA adopt a metabolic program similar to those of tumor cells with enhanced glucose utilization, lipid biosynthesis, and proliferation in hypoxic conditions. Identification of metabolic profiles underlying hyperactive T cell activation would provide a platform to test safety of immunostimulatory antibodies.


Assuntos
Antígenos CD28/imunologia , Linfócitos T CD4-Positivos/imunologia , Glicólise/imunologia , Lipogênese/imunologia , Ativação Linfocitária/imunologia , Neoplasias/metabolismo , Acetilcoenzima A/metabolismo , Anticorpos Monoclonais/imunologia , Antígenos CD28/metabolismo , Proliferação de Células , Glucose/metabolismo , Humanos , Memória Imunológica , Neoplasias/imunologia , Neoplasias/patologia , Proteínas Quinases/metabolismo , Linfócitos T Reguladores/imunologia , Células Tumorais Cultivadas
8.
Oncol Rep ; 41(5): 2945-2956, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30864716

RESUMO

Impaired antitumor immunity or induced immunosuppression in the tumor microenvironment contributes significantly to tumor progression and resistance to immunotherapy. It is becoming increasingly recognized that dynamic metabolic programming orchestrates appropriate immune responses, whereas incorrect metabolic reprogramming may underlie aberrant immune remodeling. Furthermore, pathways that control cellular metabolism and immune cell function by transcriptional and post­transcriptional mechanisms are intimately interlinked, including hypo-xia­inducible factor 1α, c­Myc and phosphatidylinositol 3­kinase/protein kinase B/mammalian target of rapamycin signaling. Immunometabolism is an emerging research field involving investigation of the interaction between immunological and metabolic processes. It is likely that high levels of nutrient competition and metabolic interplay exist between tumor cells and infiltrating immune cells in the local tumor milieu, which consequently leads to a reduction in antitumor immunity or immune cell dysfunction. Recently, a metabolic molecular mechanism responsible for the tumorigenic capacity of cluster of differentiation (CD)147, which exhibits high expression on the surface of various malignant tumor cells and is associated with tumor progression via multiple non­metabolic molecular mechanisms, was identified. The aim of the present review was to focus on the glycolytic mechanism mediated by the upregulation of CD147 in tumors and tumor­imposed metabolic restrictions on tumor­infiltrating immune cells, and the consequent immunological hyporesponsiveness. Cellular metabolism is becoming increasingly acknowledged as a key regulator of T­cell function, specification and fate, and the manipulation of metabolic programming may elucidate therapeutic options for immunological disorders and tumor immunotherapy.


Assuntos
Basigina/metabolismo , Glicólise/imunologia , Neoplasias/imunologia , Evasão Tumoral , Microambiente Tumoral/imunologia , Animais , Basigina/imunologia , Carcinogênese/imunologia , Modelos Animais de Doenças , Metabolismo Energético/imunologia , Humanos , Imunoterapia/métodos , Neoplasias/patologia , Neoplasias/terapia , Linfócitos T/imunologia , Linfócitos T/metabolismo
9.
Cell ; 177(2): 384-398.e11, 2019 04 04.
Artigo em Inglês | MEDLINE | ID: mdl-30853218

RESUMO

The signaling organelles of the innate immune system consist of oligomeric protein complexes known as supramolecular organizing centers (SMOCs). Examples of SMOCs include myddosomes and inflammasomes, which respectively induce transcription-dependent and -independent inflammatory responses. The common use of oligomeric structures as signaling platforms suggests multifunctionality, but each SMOC has a singular biochemically defined function. Here, we report that the myddosome is a multifunctional organizing center. In addition to promoting inflammatory transcription factor activation, the myddosome drives the rapid induction of glycolysis. We identify the kinase TBK1 as a myddosome component that promotes glycolysis, but not nuclear factor κB (NF-κB) activation. Synthetic immunology approaches further diversified SMOC activities, as we created interferon- or necroptosis-inducing myddosomes, inflammasomes that induce interferon responses instead of pyroptosis, and a SMOC-like nanomachine that induces interferon expression in response to a chemical ligand. These discoveries demonstrate the flexibility of immune signaling organelles, which permits the design of user-defined innate immune responses.


Assuntos
Imunidade Inata/imunologia , Imunidade Inata/fisiologia , Transdução de Sinais/imunologia , Animais , Glicólise/imunologia , Inflamassomos , Camundongos , Camundongos Endogâmicos C57BL , Enzimas Multifuncionais/imunologia , Fator 88 de Diferenciação Mieloide/metabolismo , NF-kappa B/metabolismo , Organelas/fisiologia , Proteínas Serina-Treonina Quinases/metabolismo , Receptores Toll-Like
10.
Mol Immunol ; 106: 159-169, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30623816

RESUMO

INTRODUCTION: The detoxified TLR4-ligand MPLA is a successfully used adjuvant in clinically approved vaccines. However, its capacity to activate glycolytic metabolism in mDC and the influence of MPLA-induced metabolic changes on cytokine secretion are unknown. AIM: To analyze the capacity of MPLA to activate mDC metabolism and the mechanisms contributing to MPLA-induced metabolism activation and cytokine secretion. METHODS: C57BL/6 bone-marrow-derived myeloid dendritic cells (mDCs) were stimulated with LPS or MPLA and analyzed for intracellular signaling, cytokine secretion, and metabolic state. mDC were pre-treated with rapamycin (mTOR-inhibitor), U0126, SP600125, SB202190 (MAPK kinase inhibitors), as well as dexamethasone (MAPK- and NFκB-inhibitor) and analyzed for MPLA-induced cytokine secretion and cell metabolic state. RESULTS: Stimulation of mDCs with either LPS or MPLA resulted in a pronounced, mTOR-dependent activation of glucose metabolism characterized by induction of the Warburg Effect, increased glucose consumption from the culture medium, as well as release of LDH. Compared to LPS, MPLA induced significantly lower cytokine secretion. The activation of mDC metabolism was comparable between LPS- and MPLA-stimulated mDCs. The MPLA-induced cytokine secretion could be partially inhibited using mTOR-, MAP kinase-, and NFκB-inhibitors, whereas the activation of glucose metabolism was shown to depend on both mTOR- and JNK-signaling. SUMMARY: The MPLA-induced activation of glycolytic metabolism in mouse mDC was shown to depend on a JNK-mediated activation of mTOR-signaling, while both MAPK- and NFB-signaling contributed to pro-inflammatory cytokine secretion. Understanding the mechanisms by which MPLA activates dendritic cells will both improve our understanding of its adjuvant properties and contribute to the future development and safe application of this promising adjuvant.


Assuntos
Adjuvantes Imunológicos/farmacologia , Células Dendríticas/imunologia , Glicólise/efeitos dos fármacos , Lipídeo A/análogos & derivados , MAP Quinase Quinase 4/imunologia , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Serina-Treonina Quinases TOR/imunologia , Animais , Citocinas/imunologia , Células Dendríticas/citologia , Glicólise/imunologia , Lipídeo A/farmacologia , Sistema de Sinalização das MAP Quinases/imunologia , Camundongos
11.
J Allergy Clin Immunol ; 143(1): 346-358.e6, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30096390

RESUMO

BACKGROUND: The mechanism by which natural killer (NK) cell education results in licensed NK cells with heightened effector function against missing self-targets is not known. OBJECTIVE: We sought to identify potential mechanisms of enhanced function in licensed human NK cells. METHODS: We used expanded human NK cells from killer immunoglobulin-like receptor (KIR)/HLA-genotyped donors sorted for single-KIR+ cells to generate pure populations of licensed and unlicensed NK cells. We performed proteomic and gene expression analysis of these cells before and after receptor cross-linking and performed functional and metabolic analysis before and after interference with selected metabolic pathways. We verified key findings using freshly isolated and sorted NK cells from peripheral blood. RESULTS: We confirmed that licensed human NK cells are greater in number in peripheral blood and proliferate more in vitro than unlicensed NK cells. Using high-throughput protein analysis, we found that unstimulated licensed NK cells have increased expression of the glycolytic enzyme pyruvate kinase muscle isozyme M2 and after KIR cross-linking have increased phosphorylation of the metabolic modulators p38-α and 5' adenosine monophosphate-activated protein kinase α. After cytokine expansion and activation, unlicensed NK cells depended solely on mitochondrial respiration for cytolytic function, whereas licensed NK cells demonstrated metabolic reprogramming toward glycolysis and mitochondrial-dependent glutaminolysis, leading to accumulation of glycolytic metabolites and depletion of glutamate. As such, blocking both glycolysis and mitochondrial-dependent respiration was required to suppress the cytotoxicity of licensed NK cells. CONCLUSIONS: Collectively, our data support an arming model of education in which enhanced glycolysis in licensed NK cells supports proliferative and cytotoxic capacity.


Assuntos
Glicólise/imunologia , Ativação Linfocitária/imunologia , Modelos Imunológicos , Regulação da Expressão Gênica/imunologia , Humanos , Proteômica , Receptores KIR/imunologia
12.
Methods Mol Biol ; 1862: 173-186, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30315468

RESUMO

Metabolic reprograming controlling macrophage activation and function is emerging as new regulatory circuit on shaping immune responses. Generally, lipopolysaccharides (LPS)-induced pro-inflammatory activated macrophages, known as M1 macrophages, display higher glycolysis. In contrast, interleukin-4 (IL-4)-skewed anti-inflammatory activated macrophages, known as M2 macrophages, mainly rely on oxidative phosphorylation for their bioenergetic demands. Emerging evidence reveals that these metabolic preferences further fine-tune macrophage polarization process, including signaling cascades and epigenetic reprogramming. Thus, specific nutrient microenvironments may affect inflammatory responses of macrophages by intervening these metabolic machineries. How to measure the metabolic switch of macrophages both in vitro and in vivo is an important issue for understanding immunometabolic regulations in macrophages. Here, we describe a basic protocol for examining how glutamine metabolism affects macrophage polarization by using the Extracellular Flux (XF(e)96) Analyzer (Seahorse Bioscience), which takes real-time measurements of oxidative phosphorylation and glycolysis. We also present a detailed procedure for detecting the expression of inflammatory genes in polarized macrophages under glutamine-replete or -deprived conditions.


Assuntos
Ativação de Macrófagos/imunologia , Espectrometria de Massas/métodos , Análise do Fluxo Metabólico/métodos , Metabolômica/métodos , Animais , Biomarcadores/análise , Biomarcadores/metabolismo , Células Cultivadas , Glutamina/metabolismo , Glicólise/imunologia , Lipopolissacarídeos/imunologia , Macrófagos/imunologia , Macrófagos/metabolismo , Espectrometria de Massas/instrumentação , Análise do Fluxo Metabólico/instrumentação , Redes e Vias Metabólicas/imunologia , Metabolômica/instrumentação , Camundongos , Camundongos Endogâmicos C57BL , Fosforilação Oxidativa , Consumo de Oxigênio/imunologia , Cultura Primária de Células/instrumentação , Cultura Primária de Células/métodos , Organismos Livres de Patógenos Específicos
13.
J Exp Med ; 215(12): 3180-3193, 2018 12 03.
Artigo em Inglês | MEDLINE | ID: mdl-30463876

RESUMO

Metabolic pathways such as glycolysis or oxidative phosphorylation play a key role in regulating macrophage function during inflammation and tissue repair. However, how exactly the VHL-HIF-glycolysis axis is involved in the function of tissue-resident macrophages remains unclear. Here we demonstrate that loss of VHL in myeloid cells resulted in attenuated pulmonary type 2 and fibrotic responses, accompanied by reduced eosinophil infiltration, decreased IL-5 and IL-13 concentrations, and ameliorated fiber deposition upon challenge. VHL deficiency uplifted glycolytic metabolism, decreased respiratory capacity, and reduced osteopontin expression in alveolar macrophages, which impaired the function of type 2 innate lymphoid cells but was significantly reversed by HIF1α inhibition or ablation. The up-regulated glycolysis altered the epigenetic modification of osteopontin gene, with the metabolic intermediate 3-phosphoglyceric acid as a key checkpoint controller. Thus, our results indicate that VHL acts as a crucial regulatory factor in lung inflammation and fibrosis by regulating alveolar macrophages.


Assuntos
Epigênese Genética/imunologia , Pulmão/imunologia , Macrófagos Alveolares/imunologia , Fibrose Pulmonar/imunologia , Proteína Supressora de Tumor Von Hippel-Lindau/imunologia , Animais , Glicólise/genética , Glicólise/imunologia , Inflamação/genética , Inflamação/imunologia , Inflamação/patologia , Interleucina-13/genética , Interleucina-13/imunologia , Interleucina-5/genética , Interleucina-5/imunologia , Pulmão/patologia , Macrófagos Alveolares/patologia , Camundongos , Camundongos Knockout , Fibrose Pulmonar/genética , Fibrose Pulmonar/patologia , Proteína Supressora de Tumor Von Hippel-Lindau/genética
14.
Nat Immunol ; 19(12): 1319-1329, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30397348

RESUMO

Many tumors evolve sophisticated strategies to evade the immune system, and these represent major obstacles for efficient antitumor immune responses. Here we explored a molecular mechanism of metabolic communication deployed by highly glycolytic tumors for immunoevasion. In contrast to colon adenocarcinomas, melanomas showed comparatively high glycolytic activity, which resulted in high acidification of the tumor microenvironment. This tumor acidosis induced Gprotein-coupled receptor-dependent expression of the transcriptional repressor ICER in tumor-associated macrophages that led to their functional polarization toward a non-inflammatory phenotype and promoted tumor growth. Collectively, our findings identify a molecular mechanism of metabolic communication between non-lymphoid tissue and the immune system that was exploited by high-glycolytic-rate tumors for evasion of the immune system.


Assuntos
Adenocarcinoma/imunologia , Macrófagos/imunologia , Melanoma/imunologia , Evasão Tumoral/imunologia , Microambiente Tumoral/imunologia , Acidose/imunologia , Adenocarcinoma/metabolismo , Animais , Neoplasias do Colo/imunologia , Neoplasias do Colo/metabolismo , Glicólise/imunologia , Humanos , Melanoma/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos
15.
JCI Insight ; 3(20)2018 10 18.
Artigo em Inglês | MEDLINE | ID: mdl-30333306

RESUMO

BACKGROUND: In inflammatory blood vessel diseases, macrophages represent a key component of the vascular infiltrates and are responsible for tissue injury and wall remodeling. METHODS: To examine whether inflammatory macrophages in the vessel wall display a single distinctive effector program, we compared functional profiles in patients with either coronary artery disease (CAD) or giant cell arteritis (GCA). RESULTS: Unexpectedly, monocyte-derived macrophages from the 2 patient cohorts displayed disease-specific signatures and differed fundamentally in metabolic fitness. Macrophages from CAD patients were high producers for T cell chemoattractants (CXCL9, CXCL10), the cytokines IL-1ß and IL-6, and the immunoinhibitory ligand PD-L1. In contrast, macrophages from GCA patients upregulated production of T cell chemoattractants (CXCL9, CXCL10) but not IL-1ß and IL-6, and were distinctly low for PD-L1 expression. Notably, disease-specific effector profiles were already identifiable in circulating monocytes. The chemokinehicytokinehiPD-L1hi signature in CAD macrophages was sustained by excess uptake and breakdown of glucose, placing metabolic control upstream of inflammatory function. CONCLUSIONS: We conclude that monocytes and macrophages contribute to vascular inflammation in a disease-specific and discernible pattern, have choices to commit to different functional trajectories, are dependent on glucose availability in their immediate microenvironment, and possess memory in their lineage commitment. FUNDING: Supported by the NIH (R01 AR042527, R01 HL117913, R01 AI108906, P01 HL129941, R01 AI108891, R01 AG045779 U19 AI057266, R01 AI129191), I01 BX001669, and the Cahill Discovery Fund.


Assuntos
Doença da Artéria Coronariana/imunologia , Arterite de Células Gigantes/imunologia , Glucose/metabolismo , Macrófagos/imunologia , Idoso , Idoso de 80 Anos ou mais , Artérias/imunologia , Artérias/patologia , Antígeno B7-H1/imunologia , Antígeno B7-H1/metabolismo , Células Cultivadas , Estudos de Coortes , Doença da Artéria Coronariana/sangue , Doença da Artéria Coronariana/patologia , Citocinas/imunologia , Citocinas/metabolismo , Feminino , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/imunologia , Arterite de Células Gigantes/sangue , Arterite de Células Gigantes/patologia , Glucose/imunologia , Glicólise/imunologia , Humanos , Memória Imunológica , Macrófagos/metabolismo , Masculino , Pessoa de Meia-Idade , Cultura Primária de Células
16.
J Proteome Res ; 17(11): 3780-3790, 2018 11 02.
Artigo em Inglês | MEDLINE | ID: mdl-30229649

RESUMO

Despite osteoarthritis (OA) and rheumatoid arthritis (RA) being typically age-related, their underlying etiologies are markedly different. We used 1H nuclear magnetic resonance (NMR) spectroscopy to identify differences in metabolite profiles in low volumes of OA and RA synovial fluid (SF). SF was aspirated from knee joints of 10 OA and 14 RA patients. 100 µL SF was analyzed using a 700 MHz Avance IIIHD Bruker NMR spectrometer with a TCI cryoprobe. Spectra were analyzed by Chenomx, Bruker TopSpin and AMIX software. Statistical analysis was undertaken using Metaboanalyst. 50 metabolites were annotated, including amino acids, saccharides, nucleotides and soluble lipids. Discriminant analysis identified group separation between OA and RA cohorts, with 32 metabolites significantly different between OA and RA SF (false discovery rate (FDR) < 0.05). Metabolites of glycolysis and the tricarboxylic acid cycle were lower in RA compared to OA; these results concur with higher levels of inflammation, synovial proliferation and hypoxia found in RA compared to OA. Elevated taurine in OA may indicate increased subchondral bone sclerosis. We demonstrate that quantifiable differences in metabolite abundance can be measured in low volumes of SF by 1H NMR spectroscopy, which may be clinically useful to aid diagnosis and improve understanding of disease pathogenesis.


Assuntos
Artrite Reumatoide/metabolismo , Espectroscopia de Ressonância Magnética/métodos , Metaboloma , Metabolômica/métodos , Osteoartrite/metabolismo , Líquido Sinovial/química , Idoso , Aminoácidos/química , Aminoácidos/classificação , Aminoácidos/isolamento & purificação , Artrite Reumatoide/imunologia , Artrite Reumatoide/patologia , Ciclo do Ácido Cítrico/imunologia , Estudos de Coortes , Feminino , Glicólise/imunologia , Humanos , Articulação do Joelho/imunologia , Articulação do Joelho/metabolismo , Articulação do Joelho/patologia , Lipídeos/química , Lipídeos/classificação , Lipídeos/isolamento & purificação , Masculino , Metabolômica/instrumentação , Pessoa de Meia-Idade , Nucleotídeos/química , Nucleotídeos/classificação , Nucleotídeos/isolamento & purificação , Oligossacarídeos/química , Oligossacarídeos/classificação , Oligossacarídeos/isolamento & purificação , Osteoartrite/imunologia , Osteoartrite/patologia , Líquido Sinovial/metabolismo
17.
Immunity ; 49(3): 545-559.e5, 2018 09 18.
Artigo em Inglês | MEDLINE | ID: mdl-30193848

RESUMO

Although the mammalian microbiota is well contained within the intestine, it profoundly shapes development and metabolism of almost every host organ. We questioned the range and depth of microbial metabolite penetration into the host, and how this is modulated by intestinal immunity. Chemically identical microbial and host metabolites were distinguished by stable isotope tracing from 13C-labeled live non-replicating Escherichia coli, differentiating 12C host isotopes with high-resolution mass spectrometry. Hundreds of endogenous microbial compounds penetrated 23 host tissues and fluids after intestinal exposure: subsequent 12C host metabolome signatures included lipidemia, reduced glycolysis, and inflammation. Penetrant bacterial metabolites from the small intestine were rapidly cleared into the urine, whereas induced antibodies curtailed microbial metabolite exposure by accelerating intestinal bacterial transit into the colon where metabolite transport mechanisms are limiting. Pervasive penetration of microbial molecules can cause extensive host tissue responses: these are limited by immune and non-immune intestinal mucosal adaptations to the microbiota.


Assuntos
Anticorpos/metabolismo , Microbioma Gastrointestinal/fisiologia , Glicólise/imunologia , Hiperlipidemias/imunologia , Inflamação/imunologia , Mamíferos/imunologia , Animais , Anticorpos/imunologia , Radioisótopos de Carbono/análise , Interações Hospedeiro-Patógeno , Imunidade , Cadeias Pesadas de Imunoglobulinas/genética , Espectrometria de Massas , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout
18.
Cell Metab ; 28(6): 922-934.e4, 2018 12 04.
Artigo em Inglês | MEDLINE | ID: mdl-30174303

RESUMO

T cells represent a critical effector of cell-mediated immunity. Activated T cells engage in metabolic reprogramming during effector differentiation to accommodate dynamic changes in energy demands. Here, we show that the hormone, insulin, and downstream signaling through its insulin receptor shape adaptive immune function through modulating T cell metabolism. T cells lacking insulin receptor expression (LckCre+ Insrfl/fl) show reduced antigen-specific proliferation and compromised production of pro-inflammatory cytokines. In vivo, T cell-specific insulin receptor deficiency reduces T cell-driven colonic inflammation. In a model of severe influenza infection with A/PR8 (H1N1), lack of insulin receptor on T cells curtails antigen-specific immunity to influenza viral antigens. Mechanistically, insulin receptor signaling reinforces a metabolic program that supports T cell nutrient uptake and associated glycolytic and respiratory capacities. These data highlight insulin receptor signaling as an important node integrating immunometabolic pathways to drive optimal T cell effector function in health and disease.


Assuntos
Antígenos CD/imunologia , Imunidade Celular/imunologia , Vírus da Influenza A Subtipo H1N1/imunologia , Influenza Humana/imunologia , Ativação Linfocitária/imunologia , Receptor de Insulina/imunologia , Subpopulações de Linfócitos T/imunologia , Linfócitos T/imunologia , Animais , Antígenos CD/genética , Citocinas/imunologia , Citocinas/metabolismo , Glicólise/imunologia , Humanos , Inflamação/imunologia , Inflamação/virologia , Insulina/metabolismo , Linfonodos , Camundongos , Camundongos Endogâmicos C57BL , Infecções por Orthomyxoviridae , Receptor de Insulina/genética , Transdução de Sinais , Baço , Subpopulações de Linfócitos T/citologia , Subpopulações de Linfócitos T/metabolismo , Linfócitos T/citologia , Linfócitos T/metabolismo
19.
J Exp Med ; 215(9): 2463-2476, 2018 09 03.
Artigo em Inglês | MEDLINE | ID: mdl-30115741

RESUMO

Metabolic programs are crucial for regulatory T (T reg) cell stability and function, but the underlying mechanisms that regulate T reg cell metabolism are elusive. Here, we report that lysosomal TRAF3IP3 acts as a pivotal regulator in the maintenance of T reg cell metabolic fitness. T reg-specific deletion of Traf3ip3 impairs T reg cell function, causing the development of inflammatory disorders and stronger antitumor T cell responses in mice. Excessive mechanistic target of rapamycin complex 1 (mTORC1)-mediated hyper-glycolytic metabolism is responsible for the instability of TRAF3IP3-deficient T reg cells. Mechanistically, TRAF3IP3 restricts mTORC1 signaling by recruiting the serine-threonine phosphatase catalytic subunit (PP2Ac) to the lysosome, thereby facilitating the interaction of PP2Ac with the mTORC1 component Raptor. Our results define TRAF3IP3 as a metabolic regulator in T reg cell stability and function and suggest a lysosome-specific mTORC1 signaling mechanism that regulates T reg cell metabolism.


Assuntos
Proteínas de Transporte , Glicólise , Lisossomos , Proteínas de Membrana , Transdução de Sinais , Linfócitos T Reguladores , Animais , Proteínas de Transporte/genética , Proteínas de Transporte/imunologia , Proteínas de Transporte/metabolismo , Glicólise/genética , Glicólise/imunologia , Lisossomos/genética , Lisossomos/imunologia , Lisossomos/metabolismo , Lisossomos/patologia , Proteínas de Membrana/genética , Proteínas de Membrana/imunologia , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Knockout , Proteína Regulatória Associada a mTOR/genética , Proteína Regulatória Associada a mTOR/imunologia , Proteína Regulatória Associada a mTOR/metabolismo , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismo , Linfócitos T Reguladores/patologia
20.
J Immunol ; 201(6): 1627-1632, 2018 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-30068595

RESUMO

Recent thymic emigrants (RTEs) are peripheral T cells that have most recently completed selection and thymic egress and constitute a population that is phenotypically and functionally distinct from its more mature counterpart. Ag-activated RTEs are less potent effectors than are activated mature T cells, due in part to reduced aerobic glycolysis (correctable by exogenous IL-2), which in turn impacts IFN-γ production. Mitochondria serve as nodal regulators of cell function, but their contribution to the unique biology of RTEs is unknown. In this study, we show that activated mouse RTEs have impaired oxidative phosphorylation, even in the presence of exogenous IL-2. This altered respiratory phenotype is the result of decreased CD28 signaling, reduced glutaminase induction, and diminished mitochondrial mass in RTEs relative to mature T cells. These results suggest an uncoupling whereby IL-2 tunes the rate of RTE glycolytic metabolism, whereas the unique profile of RTE mitochondrial metabolism is "hard wired."


Assuntos
Linfócitos T CD8-Positivos/imunologia , Movimento Celular/imunologia , Glicólise/imunologia , Ativação Linfocitária , Mitocôndrias/imunologia , Timo/imunologia , Animais , Antígenos CD28/genética , Antígenos CD28/imunologia , Linfócitos T CD8-Positivos/citologia , Movimento Celular/genética , Glicólise/genética , Interleucina-2/genética , Interleucina-2/imunologia , Camundongos , Camundongos Knockout , Mitocôndrias/genética , Transdução de Sinais/genética , Transdução de Sinais/imunologia , Timo/citologia
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