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1.
Life Sci ; 273: 119263, 2021 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-33636177

RESUMO

AIMS: Previous reports have found that STAT4 is involved in the epithelial-mesenchymal transition (EMT), thereby regulating the metastasis and invasion of ovarian cancer. However, the mechanisms underlying remain unclear. MAIN METHODS: We first established hypoxia-induced in vivo and in vitro models. The expression levels of signal transducer and activator of transcription 4 (STAT4), the markers of EMT and microRNA-200a (miR-200a) were assessed by western blot and qRT-PCR analysis, respectively. Through the bioinformatics analysis and luciferase assay, the relationship between miR-200a and SATA4 was performed. The gain- and loss-function experiments were performed to examine the role of miR-200a/STAT4 axis. KEY FINDINGS: The results showed that the protein level of STAT4 was significantly up-regulated in our hypoxia-exposed models, and contributed to the regulating of EMT. Besides, we found STAT4 was a direct target of miR-200a. Overexpression of miR-200a repressed the expression of STAT4, and inhibited EMT progress, whereas the silencing of miR-200a promoted the STAT4-mediated EMT regulation both in vitro and in vivo. SIGNIFICANCE: Our results provided a potential molecular mechanism by which miR-200a involved in hypoxia-induced metastasis and invasion in ovarian cancer, suggesting a possible target for the treatment of ovarian cancer.


Assuntos
Biomarcadores Tumorais/metabolismo , Regulação Neoplásica da Expressão Gênica , Hipóxia/fisiopatologia , MicroRNAs/genética , Neoplasias Ovarianas/patologia , Fator de Transcrição STAT4/metabolismo , Animais , Apoptose , Biomarcadores Tumorais/genética , Estudos de Casos e Controles , Movimento Celular , Proliferação de Células , Transição Epitelial-Mesenquimal , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Pessoa de Meia-Idade , Neoplasias Ovarianas/genética , Neoplasias Ovarianas/metabolismo , Prognóstico , Fator de Transcrição STAT4/genética , Transdução de Sinais , Taxa de Sobrevida , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de Xenoenxerto
2.
J Leukoc Biol ; 107(4): 663-671, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32017227

RESUMO

This study tests the hypothesis that activation of MAPK by physiologically relevant concentrations of IL-33 contributes to enhanced cytokine expression by IL-12 stimulated human NK cells. While IL-33 canonically triggers type 2 cytokine responses, this cytokine can also synergize with type 1 cytokines like IL-12 to provoke IFN-γ. We show that picogram concentrations of IL-12 and IL-33 are sufficient to promote robust secretion of IFN-γ by human NK cells that greatly exceeds resposes to either cytokine alone. Nanogram doses of IL-33, potentially consistent with levels in tissue microenvironments, synergize with IL-12 to induce secretion of additional cytokines, including TNF and GM-CSF. IL-33-induced activation of the p38 MAPK pathway in human NK cells is crucial for enhanced release of IFN-γ and TNF in response to IL-12. Mechanistically, IL-33-induced p38 MAPK signaling enhances stability of IFNG transcripts and triggers A disintegrin and metalloproteinase domain 17 (ADAM17) mediated cleavage of TNF from the cell surface. These data support our hypothesis and suggest that altered sensitivity of NK cells to IL-12 in the presence of IL-33 may have important consequences in diseases associated with mixed cytokine milieus, like asthma and chronic obstructive pulmonary disease.


Assuntos
Citocinas/metabolismo , Interleucina-33/metabolismo , Células Matadoras Naturais/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo , Proteína ADAM17/metabolismo , Linhagem Celular , Humanos , Interferon gama/genética , Interferon gama/metabolismo , Proteína 1 Semelhante a Receptor de Interleucina-1/metabolismo , Interleucina-12/metabolismo , Fosforilação , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fator de Transcrição STAT4/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores
3.
Cell Rep ; 30(4): 984-996.e4, 2020 01 28.
Artigo em Inglês | MEDLINE | ID: mdl-31995767

RESUMO

The induction of broadly neutralizing antibodies (bnAbs) is highly desired for an effective vaccine against HIV-1. Typically, bnAbs develop in patients with high viremia, but they can also evolve in some untreated HIV-1 controllers with low viral loads. Here, we identify a subgroup of neutralizer-controllers characterized by myeloid DCs (mDCs) with a distinct inflammatory signature and a superior ability to prime T follicular helper (Tfh)-like cells in an STAT4-dependent fashion. This distinct immune profile is associated with a higher frequency of Tfh-like cells in peripheral blood (pTfh) and an enrichment for Tfh-defining genes in circulating CD4+ T cells. Correspondingly, monocytes from this neutralizer controller subgroup upregulate genes encoding for chemotaxis and inflammation, and they secrete high levels of IL-12 in response to TLR stimulation. Our results suggest the existence of multi-compartment immune networks between mDCs, Tfh, and monocytes that may facilitate the development of bnAbs in a subgroup of HIV-1 controllers.


Assuntos
Anticorpos Neutralizantes/imunologia , Linfócitos T CD4-Positivos/imunologia , Células Dendríticas/imunologia , Anticorpos Anti-HIV/imunologia , Infecções por HIV/imunologia , HIV-1/imunologia , Monócitos/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Adulto , Idoso , Linfócitos B/imunologia , Linfócitos B/metabolismo , Linfócitos T CD4-Positivos/metabolismo , Linhagem Celular , Sobrevivência Celular/imunologia , Células Dendríticas/metabolismo , Feminino , Regulação da Expressão Gênica/imunologia , Humanos , Interleucina-12/metabolismo , Masculino , Pessoa de Meia-Idade , Monócitos/metabolismo , RNA-Seq , Fator de Transcrição STAT4/genética , Fator de Transcrição STAT4/metabolismo , Linfócitos T Auxiliares-Indutores/metabolismo , Células Th1/imunologia , Células Th1/metabolismo
4.
Biomed Res Int ; 2019: 1703842, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31871930

RESUMO

Systemic lupus erythematosus (SLE) is characterized by systemic end-organ damage. We investigated the involvement of IRF5, TLR-7, MECP2, STAT4, and TNFSF4 genes and TNF-α, IFN-γ, IL-2, IL-12, IL-6, and IL-10 cytokines in SLE pathogenesis and in organ damage in Jordanian patients. Blood was collected from 51 patients and 50 controls. Expression levels of SLE genes in PBMCs and cytokine levels were determined using RT-PCR and ELISA, respectively. Expression levels of all genes and levels of TNF-α, IL-12, IL-6, and IL-10 were higher in SLE patients than those in controls (p < 0.05), whereas IL-2 level was lower. High STAT4 (α), TNFSF4, and IL-10 levels correlated with cardiovascular damage, and high MECP2 (α) and TNF-α correlated with renal damage. Pulmonary and musculoskeletal damages correlated with high levels of TNFSF4. We concluded that STAT4 and TNFSF4 genes with TNF-α and IL-10 cytokines could be used as biomarkers to assess SLE activity and manage treatment.


Assuntos
Citocinas/metabolismo , Fatores Reguladores de Interferon/metabolismo , Lúpus Eritematoso Sistêmico/metabolismo , Proteína 2 de Ligação a Metil-CpG/metabolismo , Ligante OX40/metabolismo , Fator de Transcrição STAT4/metabolismo , Receptor 7 Toll-Like/metabolismo , Adulto , Biomarcadores/sangue , Citocinas/sangue , Feminino , Regulação da Expressão Gênica , Humanos , Fatores Reguladores de Interferon/genética , Interferon gama/metabolismo , Interleucina-10/metabolismo , Interleucina-12/metabolismo , Interleucina-2/metabolismo , Interleucina-6/metabolismo , Jordânia , Lúpus Eritematoso Sistêmico/genética , Masculino , Proteína 2 de Ligação a Metil-CpG/genética , Ligante OX40/genética , RNA Mensageiro/metabolismo , Fator de Transcrição STAT4/genética , Receptor 7 Toll-Like/genética , Fator de Necrose Tumoral alfa/metabolismo
5.
Pharm Biol ; 57(1): 744-752, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31679431

RESUMO

Context: Liuweibuqi (LWBQ) capsule has been reported to influence symptoms of patients with chronic obstructive pulmonary disease (COPD); however, specific function of LWBQ capsules in COPD with lung-qi deficiency syndrome remains elusive.Objective: This study investigates effect of LWBQ capsules on STAT4/STAT6 and MMP-9/TIMP-1 expression and pulmonary function in stable COPD with lung-qi deficiency syndrome.Materials and methods: Totally, 429 patients diagnosed with stable COPD and lung-qi deficiency syndrome were treated with starch capsules (each time for 9 capsules), or different doses: low (each dose for 8 capsules and 1 LWBQ capsules), medium (each time for 6 capsules and 3 LWBQ capsules), or high (each time for 9 LWBQ capsules) of LWBQ capsules for 30 days, 3 times a day. Forced expiratory volume in 1 s (FEV1), forced vital capacity (FVC), FEV1/FVC% and DLco%pred were evaluated by pulmonary function meter. STAT4/STAT6 and MMP-9/TIMP-1 expression was assessed by RT-qPCR and western blot analysis, and serum concentrations of IL-4, IFN-γ and IL-6 by ELISA.Results: Spearman rank correlation analysis and ROC curve showed that STAT4/STAT6 and MMP-9/TIMP-1 affected pulmonary functions and curative effect of stable COPD with lung-qi deficiency syndrome. After LWBQ capsule treatment, FEV1, FVC, FEV1/FVC% and DLco%pred elevated; STAT4/STAT6, MMP-9/TIMP-1, IFN-γ and IL-6 expression declined whereas IL-4 expression increased (p < 0.05). Logistic regression analysis demonstrated that FEV1/FVC was negatively correlated with STAT4/STAT6 and MMP-9/TIMP-1 expression in COPD patients.Conclusions: LWBQ capsules play a beneficial role in pulmonary function of stable COPD with lung-qi deficiency syndrome via STAT4/STAT6 and MMP-9/TIMP-1.


Assuntos
Medicamentos de Ervas Chinesas/farmacologia , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico , Qi , Cápsulas , Medicamentos de Ervas Chinesas/administração & dosagem , Feminino , Volume Expiratório Forçado , Humanos , Masculino , Metaloproteinase 9 da Matriz/metabolismo , Pessoa de Meia-Idade , Doença Pulmonar Obstrutiva Crônica/patologia , Testes de Função Respiratória , Fator de Transcrição STAT4/metabolismo , Fator de Transcrição STAT6/metabolismo , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Resultado do Tratamento , Capacidade Vital
6.
Sci Rep ; 9(1): 13991, 2019 09 30.
Artigo em Inglês | MEDLINE | ID: mdl-31570752

RESUMO

CD4+ T follicular helper (TFH) cells provide help to B cells and promote antibody-mediated immune responses. Increasing evidence supports the existence of TFH populations that secrete cytokines typically associated with the effector functions of other CD4+ T cell subsets. These include T helper 1 (TH1)-biased TFH (TFH1) cells that have recognized roles in both immune responses to pathogens and also the pathogenesis of autoimmune disease. Given their apparent importance to human health, there is interest in understanding the mechanisms that regulate TFH1 cell formation and function. However, their origin and the molecular requirements for their differentiation are unclear. Here, we describe a population of murine TH1-derived, TFH1-like cells that express the chemokine receptor Cxcr3 and produce both the TH1 cytokine interferon-γ and the TFH-associated cytokine interleukin-21 (IL-21). Furthermore, these TFH1-like cells promote B cell activation and antibody production at levels indistinguishable from conventional IL-6-derived TFH-like cells. Regarding their regulatory requirements, we find that IL-12 signaling is necessary for the differentiation and function of this TFH1-like cell population. Specifically, IL-12-dependent activation of STAT4, and unexpectedly STAT3, promotes increased expression of IL-21 and the TFH lineage-defining transcription factor Bcl-6 in TFH1-like cells. Taken together, these findings provide insight into the potential origin and differentiation requirements of TFH1 cells.


Assuntos
Interleucina-12/metabolismo , Transdução de Sinais , Células Th1/fisiologia , Animais , Diferenciação Celular , Citometria de Fluxo , Regulação da Expressão Gênica , Interferon gama/metabolismo , Interleucina-12/fisiologia , Interleucinas/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Reação em Cadeia da Polimerase , Fator de Transcrição STAT3/metabolismo , Fator de Transcrição STAT4/metabolismo , Células Th1/metabolismo
7.
EMBO Rep ; 20(11): e48647, 2019 11 05.
Artigo em Inglês | MEDLINE | ID: mdl-31549795

RESUMO

The transcription factors STAT3 and STAT4 are essential for lymphocyte differentiation and function. Interleukin (IL)-17 producing γδ T (γδT17) cells are innate lymphocytes important for anti-bacterial and inflammatory responses at barrier surfaces. Herein, we examine the role of STAT3 and STAT4 in regulating the homeostasis, activation, and pathogenicity of γδT17 cells. We show that STAT3 sustains γδT17 numbers in the skin but not in the lymph nodes, while STAT4 deficiency does not affect their homeostasis. Similarly, STAT3 but not STAT4 is essential for IL-23-induced IL-22 production by γδT17 cells. Concomitantly, mice lacking STAT3 expression in γδT17 cells develop significantly reduced psoriasis-like inflammation. STAT3-deficient γδT17 cells fail to expand and to upregulate IL-17A, IL-17F, and IL-22 in response to psoriatic stimuli. Although STAT4-deficient animals develop psoriasis-like disease, γδT17 cells in these mice are defective in IL-17F production. Collectively, our data demonstrate for the first time a critical role for STAT3 in orchestrating the homeostasis and pathogenicity of γδT17 cells and provide evidence for the requirement of STAT4 for optimal cytokine responses during inflammation.


Assuntos
Dermatite/etiologia , Dermatite/metabolismo , Receptores de Antígenos de Linfócitos T gama-delta/metabolismo , Fator de Transcrição STAT3/metabolismo , Fator de Transcrição STAT4/metabolismo , Células Th17/imunologia , Células Th17/metabolismo , Animais , Biomarcadores , Biópsia , Citocinas/metabolismo , Modelos Animais de Doenças , Suscetibilidade a Doenças , Imunomodulação , Imunofenotipagem , Mediadores da Inflamação/metabolismo , Contagem de Linfócitos , Camundongos , Psoríase/etiologia , Psoríase/metabolismo
8.
Mol Cell Endocrinol ; 498: 110541, 2019 12 01.
Artigo em Inglês | MEDLINE | ID: mdl-31415795

RESUMO

MicroRNAs (miRNAs) are small RNAs without protein-coding functions that negatively regulate target genes and play important roles in physiological and pathological processes. The aim of this work was to reveal a novel miRNA/gene pathway in diabetic retinopathy (DR). A microarray was used to screen miRNAs in samples from nondiabetic controls and patients with DR, and miR-223-3p was screened as a potential candidate. Quantitative real-time PCR (qRT-PCR) revealed that the level of miR-223-3p was frequently overexpressed in DR samples and human retinal endothelial cells (hRECs) in hyperglycemia, but it was decreased in hyperglycemia after the addition of transthyretin (TTR). In addition, according to cell proliferation, tube formation, and wound healing assays, the downregulation of miR-223-3p suppressed cell migration and proliferation, whereas miR-223-3p upregulation showed the opposite effects. Furthermore, luciferase assays identified F-box and WD repeat domain-containing 7 (FBXW7) as a target mRNA of miR-223-3p. High glucose conditions facilitated the recruitment of signal transducer and activator of transcription 4 (STAT4) and promoted the transcription of miR-223-3p. In hRECs, in a hyperglycemic environment, TTR inhibited STAT4 expression, downregulated the level of miR-223-3p, and finally promoted FBXW7 expression. This study found a novel mechanism whereby TTR might affect neovascularization through a newly identified STAT4/miR-223-3p/FBXW7 cascade in DR.


Assuntos
Retinopatia Diabética/complicações , Proteína 7 com Repetições F-Box-WD/metabolismo , Hiperglicemia/fisiopatologia , MicroRNAs/genética , Neovascularização Patológica/patologia , Pré-Albumina/metabolismo , Fator de Transcrição STAT4/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Movimento Celular , Proliferação de Células , Retinopatia Diabética/metabolismo , Retinopatia Diabética/patologia , Proteína 7 com Repetições F-Box-WD/genética , Feminino , Regulação da Expressão Gênica , Humanos , Hiperglicemia/metabolismo , Masculino , Pessoa de Meia-Idade , Neovascularização Patológica/etiologia , Neovascularização Patológica/metabolismo , Pré-Albumina/genética , Retina/metabolismo , Retina/patologia , Fator de Transcrição STAT4/genética , Transdução de Sinais
9.
Am J Physiol Heart Circ Physiol ; 317(3): H531-H540, 2019 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-31225989

RESUMO

As an inflammatory disease afflicting the heart muscle, autoimmune myocarditis (AM) represents one of the foremost causes of heart failure. Accumulating evidence has implicated microRNAs (miRNAs) in the process of inflammation and autoimmunity. Hence, the current study aimed to investigate the mechanism by which miR-141-3p influences experimental AM (EAM). An EAM mouse model was established using 6-wk old male BALB/c mice, after which the expression of miR-141-3p and STAT4 was measured. Gain-of-function and loss-of-function investigations were performed to identify the functional role of miR-141-3p and STAT4 in EAM. Heart weight-to-body weight ratio, cardiac function, and degree of inflammation, as well as the levels of inflammation factors (IFN-γ, TNF-α, IL-2, IL-6, and IL-17) in the serum were detected. STAT4 was subsequently verified to be upregulated, and miR-141-3p was downregulated in the EAM mice. Furthermore, the overexpression of miR-141-3p or silencing of STAT4 was observed to reduce the heart weight-to-body weight ratio of EAM mice and improve cardiac function, while alleviating the degree of inflammatory cell infiltration in the myocardial tissue. Meanwhile, the overexpression of miR-141-3p was identified to diminish serum inflammatory factor levels by downregulating STAT4. Additionally, miR-141-3p could bind to STAT4 to downregulate its expression, ultimately mitigating inflammation and inducing an anti-inflammatory effect in EAM mice. Taken together, upregulation of miR-141-3p alleviates the inflammatory response in EAM mice by inhibiting STAT4, providing a promising intervention target for the molecular treatment of AM.NEW & NOTEWORTHY miR-141-3p is poorly expressed, and STAT4 is upregulated in experimental autoimmune myocarditis (EAM) mice. Overexpressing miR-141-3p inhibits EAM. miR-141-3p binds to and suppresses STAT4 expression. miR-141-3p overexpression inhibits inflammatory factors by downregulating STAT4. This study provides new insights into the treatment of autoimmune myocarditis.


Assuntos
Doenças Autoimunes/prevenção & controle , MicroRNAs/metabolismo , Miocardite/prevenção & controle , Miocárdio/metabolismo , RNA Interferente Pequeno/administração & dosagem , Terapêutica com RNAi , Fator de Transcrição STAT4/metabolismo , Animais , Antagomirs/administração & dosagem , Doenças Autoimunes/genética , Doenças Autoimunes/imunologia , Doenças Autoimunes/metabolismo , Citocinas/sangue , Citocinas/genética , Modelos Animais de Doenças , Redes Reguladoras de Genes , Mediadores da Inflamação/sangue , Masculino , Camundongos Endogâmicos BALB C , MicroRNAs/genética , Miocardite/genética , Miocardite/imunologia , Miocardite/metabolismo , Miocárdio/imunologia , Miosinas , Interferência de RNA , Fator de Transcrição STAT4/genética
10.
Cytokine ; 120: 251-257, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31146247

RESUMO

IL-12 is a key cytokine for the promotion of CD4+ T cells differentiation to type 1 helper T cells. IL-12 is a heterodimer (IL-12p70) consisting of p40 and p35 subunits, and is mainly secreted from activated antigen-presenting cells, such as macrophages and dendritic cells (DCs). In this study, we found that activated mouse bone marrow-derived DCs (BMDCs) produced a p40 splice variant form mRNA in addition to the conventional p40 mRNA. This p40 variant mRNA was produced by alternative splicing in exon 5, and possessed a premature stop codon. As a result, the p40 variant protein contained 157 amino acids of the N-terminal part of p40 and an additional 10 novel amino acids. When the p40 variant was expressed in HEK-293T cells, it was not secreted from the cells. To investigate the function of the p40 variant, it was co-expressed with p40 and/or p35. The p40 variant did not affect the secretion of IL-12p40 or IL-12p70, or the function of the secreted p70. In contrast, the secretion of IL-12p80, a homodimeric IL-12 with two p40 subunits, was significantly decreased when the p40 variant was expressed. This new splicing variant p40 may act to fine-tune the function of IL-12p80.


Assuntos
Processamento Alternativo/genética , Subunidade p40 da Interleucina-12/genética , Interleucina-12/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Éxons/genética , Células HEK293 , Humanos , Interleucina-12/química , Subunidade p40 da Interleucina-12/química , Cinética , Camundongos Endogâmicos C57BL , Fosforilação , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fator de Transcrição STAT4/metabolismo
11.
Front Immunol ; 10: 913, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31080452

RESUMO

While IL-12 plays a key role in differentiation of protective CD4+ Th1 response, little is known about mechanisms whereby IL-12 differentiates other T-cell populations. Published studies suggest that predominant Vγ2Vδ2 T cells in humans/nonhuman primates (NHP) are a fast-acting T-cell subset, with capacities to rapidly expand and produce Th1 and cytotoxic cytokines in response to phosphoantigen (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMBPP) produced by Mycobacterium tuberculosis (Mtb) or others. However, whether IL-12 signaling pathway mediates fast-acting and Th1 or anti-microbial features of Vγ2Vδ2 T cells remains poorly defined. Here, we show that IL-12, but not other IL-12 family members IL-27/IL-35, apparently expanded HMBPP-activated Vγ2Vδ2 T cells. Although IL-12 and IL-2 similarly expanded HMBPP-activated Vγ2Vδ2 T-cell clones, the IL-12-induced expansion did not require endogenous IL-2 or IL-2 co-signaling during HMBPP + IL-12 co-treatment. IL-12-induced expansion of Vγ2Vδ2 T cells required the PI3K/AKT and STAT4 activation pathways and endogenous TNF-α signaling but did not involve p38/MAPK or IFN-γ signals. IL-12-expanded Vγ2Vδ2 T cells exhibited central/effector memory phenotypes and differentiated into polyfunctional effector cell subtypes which expressed TBX21/T-bet, antimicrobial cytokines IFN-γ, TNF-α, GM-CSF, and cytotoxic granule molecules. Furthermore, the IL-12-expanded Vγ2Vδ2 T cells inhibited the growth of intracellular mycobacteria in IFN-γ- or TNF-α-dependent fashion. Our findings support the concept that IL-12 drives early development of fast-acting Vγ2Vδ2 T effector cells in antimicrobial immune responses.


Assuntos
Interleucina-12/imunologia , Linfócitos Intraepiteliais/imunologia , Mycobacterium tuberculosis/imunologia , Subpopulações de Linfócitos T/imunologia , Células Th1/imunologia , Diferenciação Celular/imunologia , Células Cultivadas , Humanos , Ativação Linfocitária/efeitos dos fármacos , Organofosfatos/farmacologia , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fator de Transcrição STAT4/metabolismo , Transdução de Sinais/imunologia , Fator de Necrose Tumoral alfa/metabolismo
12.
EBioMedicine ; 43: 380-391, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30992245

RESUMO

BACKGROUND: Systemic inflammation induced by sterile or infectious insults is associated with an enhanced susceptibility to life-threatening opportunistic, mostly bacterial, infections due to unknown pathogenesis. Natural killer (NK) cells contribute to the defence against bacterial infections through the release of Interferon (IFN) γ in response to Interleukin (IL) 12. Considering the relevance of NK cells in the immune defence we investigated whether the function of NK cells is disturbed in patients suffering from serious systemic inflammation. METHODS: NK cells from severely injured patients were analysed from the first day after the initial inflammatory insult until the day of discharge in terms of IL-12 receptor signalling and IFN-γ synthesis. FINDINGS: During systemic inflammation, the expression of the IL-12 receptor ß2 chain, phosphorylation of signal transducer and activation 4, and IFN-γ production on/in NK cells was impaired upon exposure to Staphylococcus aureus. The profound suppression of NK cells developed within 24 h after the initial insult and persisted for several weeks. NK cells displayed signs of exhaustion. Extrinsic changes were mediated by the early and long-lasting presence of growth/differentiation factor (GDF) 15 in the circulation that signalled through the transforming growth factor ß receptor I and activated Smad1/5. Moreover, the concentration of GDF-15 in the serum inversely correlated with the IL-12 receptor ß2 expression on NK cells and was enhanced in patients who later acquired septic complications. INTERPRETATION: GDF-15 is associated with the development of NK cell dysfunction during systemic inflammation and might represent a novel target to prevent nosocomial infections. FUND: The study was supported by the Department of Orthopaedics and Trauma Surgery, University Hospital Essen.


Assuntos
Antígeno CD56/metabolismo , Infecção Hospitalar/etiologia , Infecção Hospitalar/metabolismo , Fator 15 de Diferenciação de Crescimento/sangue , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Adulto , Biomarcadores , Comorbidade , Infecção Hospitalar/sangue , Infecção Hospitalar/diagnóstico , Feminino , Humanos , Imunofenotipagem , Mediadores da Inflamação/metabolismo , Interferon gama/metabolismo , Interleucina-12/metabolismo , Masculino , Pessoa de Meia-Idade , Fosforilação , Receptores de Interleucina-12/metabolismo , Fator de Transcrição STAT4/metabolismo , Índice de Gravidade de Doença , Transdução de Sinais , Síndrome de Resposta Inflamatória Sistêmica/sangue , Síndrome de Resposta Inflamatória Sistêmica/etiologia , Síndrome de Resposta Inflamatória Sistêmica/metabolismo
13.
Inflammation ; 42(4): 1179-1189, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30848408

RESUMO

Signal transducer and activator of transcription 4 (STAT4) has been implicated in the progression of myocarditis. The aim of the current study was to investigate the role by which STAT4 influences autoimmune myocarditis in an attempt to identify a theoretical therapeutic perspective for the condition. After successful establishment of an autoimmune myocarditis rat model, the expression patterns of STAT4, NF-κB pathway-related genes, Th1 inflammatory cytokines (IFN-γ and IL-2), and Th2 inflammatory cytokines (IL-6 and IL-10) were subsequently determined. The rats with autoimmune myocarditis were treated with oe-STAT4 or sh-STAT4 lentiviral vectors to evaluate the role of STAT4 in autoimmune myocarditis, or administrated with 1 mL 10 µmol/L of BAY11-7082 (the NF-κB pathway inhibitor) via tail vein to investigate the effect of the NF-κB pathway on autoimmune myocarditis. Finally, cell apoptosis was evaluated. The serum levels of IFN-γ and IL-2, extent of IκBα and P65 phosphorylation, and the expression of STAT4 were elevated, while the serum levels of IL-6 and IL-10 as well as the expression of IκBα were reduced among the rats with autoimmune myocarditis, which was accompanied by an increase in the apoptotic cells. More importantly, the silencing of STAT4 or the inhibition of the NF-κB pathway was detected to result in a decrease in the serum levels of IFN-γ and IL-2 and an elevation of the serum levels of IL-6 and IL-10, and inhibited myocardial cell apoptosis in rats with autoimmune myocarditis. Moreover, STAT4 silencing was also observed to decrease the extent of IκBα and P65 phosphorylation while acting to elevate the expression of IκBα. Taken together, silencing of STAT4 could hinder the progression of autoimmune myocarditis by balancing the expression of Th1/Th2 inflammatory cytokines via the NF-κB pathway, which may provide a novel target for experimental autoimmune myocarditis (EAM) treatment.


Assuntos
Miocardite/imunologia , NF-kappa B/antagonistas & inibidores , Fator de Transcrição STAT4/fisiologia , Células Th1/imunologia , Células Th2/imunologia , Animais , Apoptose , Doenças Autoimunes , Citocinas/sangue , Progressão da Doença , Miocardite/prevenção & controle , NF-kappa B/metabolismo , Ratos , Fator de Transcrição STAT4/antagonistas & inibidores , Fator de Transcrição STAT4/metabolismo
14.
Iran J Immunol ; 16(1): 71-83, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30864557

RESUMO

BACKGROUND: STAT4 is a transcription factor that plays a role in various cytokine signaling pathways and in T cell subsets differentiation. Several studies have reported STAT4 gene polymorphism in association with various autoimmune diseases. OBJECTIVE: To evaluated the association between STAT4 rs7574865 SNP and RA risk by meta-analysis. METHODS: Two major databases, namely Scopus and PubMed, were searched to find studies investigating the STAT4 polymorphism and RA in different populations up to November 2017. Association between STAT4 polymorphism and RA were analyzed using pooled odds ratio (OR) and their corresponding 95% CI. RESULTS: In this meta-analysis, 21 population studies (16 papers) comprising 15,732 cases and 15641 healthy subjects evaluating the STAT4 gene rs7574865 SNP were included based on inclusion criteria. Herein, we found a significant positive association between minor T allele as well as different genotypes with the risk of RA. CONCLUSIONS: In summary, this study revealed an association between STAT4 gene rs7574865 SNP and risk of RA.


Assuntos
Artrite Reumatoide/etiologia , Predisposição Genética para Doença , Polimorfismo de Nucleotídeo Único , Fator de Transcrição STAT4/genética , Alelos , Artrite Reumatoide/metabolismo , Artrite Reumatoide/patologia , Frequência do Gene , Estudos de Associação Genética , Genótipo , Humanos , Razão de Chances , Viés de Publicação , Fator de Transcrição STAT4/metabolismo
15.
J Leukoc Biol ; 106(3): 733-747, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-30861206

RESUMO

Host-pathogen interactions in tuberculosis (TB) should be studied at the disease sites because Mycobacterium tuberculosis (M.tb) is predominantly contained in local tissue lesions. T-cell immune responses are required to mount anti-mycobacterial immunity. However, T-cell immune responses modulated by programmed cell death protein 1 (PD-1) during tuberculosis pleurisy (TBP) remains poorly understood. We selected the pleural fluid mononuclear cells (PFMCs) from TBP and PBMCs from healthy donors (HD), and characterized PD-1-expresing T-cell phenotypes and functions. Here, we found that the PFMCs exhibited increases in numbers of PD-1-expressing CD4+ and CD8+ T cells, which preferentially displayed polarized effector memory phenotypes. The M.tb-specific Ag stimulation increased CD4+ PD-1+ and CD8+ PD-1+ T cells, which is in direct correlation with IFN-γ production and PD-L1+ APCs in PFMCs of these individuals. Moreover, blockage of PD-1/PD-L1 pathway enhanced the percentage of IFN-γ+ T cells, demonstrating that the PD-1/PD-L1 pathway played a negative regulation in T cell effector functions. Furthermore, CD4+ PD-1+ and CD8+ PD-1+ T-cell subsets showed greater memory phenotype, activation, and effector functions for producing Th1 cytokines than PD-1- counterparts. Thus, these PD-1+ T cells were not exhausted but appear to be central to maintaining Ag-specific effector. IL-12, a key immunoregulatory cytokine, enhanced the expression of PD-1 and restored a strong IFN-γ response through selectively inducing the phosphorylation of STAT4 in CD4+ PD-1+ T-bet+ and CD8+ PD-1+ T-bet+ T cells. This study therefore uncovered a previously unknown mechanism for T-cell immune responses regulated by PD-1, and may have implications for potential immune intervention in TBP.


Assuntos
Polaridade Celular , Memória Imunológica , Mycobacterium tuberculosis/fisiologia , Receptor de Morte Celular Programada 1/metabolismo , Linfócitos T/imunologia , Tuberculose Pleural/imunologia , Tuberculose Pleural/microbiologia , Adulto , Idoso , Células Apresentadoras de Antígenos/metabolismo , Antígeno B7-H1/metabolismo , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Citocinas/metabolismo , Feminino , Humanos , Interferon gama/metabolismo , Ligantes , Ativação Linfocitária/imunologia , Masculino , Pessoa de Meia-Idade , Fosforilação , Fator de Transcrição STAT4/metabolismo , Células Th1/imunologia , Regulação para Cima , Adulto Jovem
16.
BMC Infect Dis ; 19(1): 52, 2019 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-30642265

RESUMO

BACKGROUND: Leprosy is an ideal human disease to study T cell regulation as patients show correlation between cytokine skewed Th1-Th2 responses and clinical forms of the disease. The Role of transcription factors on the modulation of Th1 and Th2 responses by M. leprae antigens has not been adequately studied. In the present study, we studied the effect of M. leprae antigens on transcription factors STAT-4, STAT-6 and CREB and their correlation with Th1/Th2 cell mediated immune responses in leprosy. METHODS: Leprosy patients of both categories of tuberculoid leprosy (BT/TT) and lepromatous leprosy (BL/LL) were selected from the OPD of NJ1L & OMD, (ICMR), Agra and healthy individuals (H) were chosen from the staff and students working in the institute. Peripheral blood mononuclear cells (PBMCs) of the study subjects were stimulated with M. leprae antigens (WCL, MLSA, and PGL-1). Sandwich ELISA was done in the culture supernatants of healthy and leprosy patients to detect IL-4, IL-10 and IFN-γ. Further, expression of IFN-γ and IL-4 and activation of STAT4, STAT6 and CREB transcription factors in CD4+ T cell with or without stimulation of M. leprae antigens was investigated by flow cytometry. RESULTS: Lepromatous leprosy patients showed significantly lower IFN-γ and higher IL-4 levels in culture supernatant and significantly low expression of IFN-γ and higher expression of IL-4 by CD4+ T cells than healthy individuals with or without antigenic stimulation. Antigenic stimulation significantly increased IL-10 in BL/LL patients but not in BT/TT patients or healthy individuals. PGL-1 stimulation led to significantly higher activation of STAT-6 in BT/TT and BL/LL patients in comparison to healthy individuals. All the three antigens led to activation of CREB in healthy and BT/TT patients but not in BL/LL patients. CONCLUSION: Our findings show that M. leprae antigens differentially modulate activation of T cell transcription factors STAT-4/STAT-6 and CREB. These transcription factors are well known to regulate Th1 and Th2 mediated immune response which in turn could play vital role in the clinical manifestations of leprosy. These observations may help to determine how these T cell transcription factors affect the development of immune dysfunction and whether these new pathways have a role in immunomodulation in intracellular diseases like leprosy and TB.


Assuntos
Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Hanseníase/imunologia , Mycobacterium leprae/imunologia , Fator de Transcrição STAT4/metabolismo , Fator de Transcrição STAT6/metabolismo , Adulto , Antígenos de Bactérias/farmacologia , Linfócitos T CD4-Positivos/efeitos dos fármacos , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Estudos de Casos e Controles , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/imunologia , Citocinas/metabolismo , Humanos , Hanseníase/metabolismo , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/microbiologia , Pessoa de Meia-Idade , Mycobacterium leprae/patogenicidade , Fator de Transcrição STAT4/imunologia , Fator de Transcrição STAT6/imunologia , Células Th1/efeitos dos fármacos , Células Th1/imunologia , Células Th1/metabolismo , Células Th2/efeitos dos fármacos , Células Th2/imunologia , Células Th2/metabolismo
17.
J Cell Biochem ; 120(6): 9147-9158, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30582204

RESUMO

Interstitial cystitis (IC) is a heterogeneous syndrome with unknown etiology, and microRNAs (miRs) were found to be involved in IC. In our study, we aim to explore the role of miR-132 in the inflammatory response and detrusor fibrosis in IC through the Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway in rat models. A rat model of IC was established and treated with the miR-132 mimic, miR-132 inhibitor, and/or JAK-STAT signaling pathway inhibitor AG490. Enzyme-linked immunosorbent assay was applied to measure the expression of interleukin (IL)-6, IL-10, interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α), and intercellular adhesion molecule-1 (ICAM-1). The urodynamic test was performed to assess urodynamic parameters, and reverse transcription quantitative polymerase chain reaction and Western blot analysis for the expression of miR-132, STAT4, suppressors of cytokine signaling 3 (SOCS3), JAK2, vascular endothelial growth factor (VEGF), IFN-γ, and TNF-α. IC rats treated with miR-132 inhibitor and AG490 had decreased collagen fiber, inflammatory cell infiltration, and mast cells, lower expression of IL-6, IL-10, IFN-γ, TNF-α, ICAM-1, collagens I and III, and alleviated urodynamic parameters and decreased expression of STAT4, VEGF, JAK2, IFN-γ, TNF-α, and increased expression of SOCS3. Taken together, our data indicate that downregulation of miR-132 alleviates inflammatory response and detrusor fibrosis in IC via the inhibition of the JAK-STAT signaling pathway.


Assuntos
Cistite Intersticial/metabolismo , Inflamação/metabolismo , Janus Quinases/metabolismo , MicroRNAs/metabolismo , Animais , Cistite Intersticial/tratamento farmacológico , Feminino , Humanos , Imuno-Histoquímica , Técnicas In Vitro , Inflamação/tratamento farmacológico , Janus Quinase 2/metabolismo , MicroRNAs/genética , Ratos , Ratos Sprague-Dawley , Fator de Transcrição STAT4/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/genética , Fator de Necrose Tumoral alfa/metabolismo , Tirfostinas/uso terapêutico , Fator A de Crescimento do Endotélio Vascular/metabolismo
18.
Exp Mol Pathol ; 107: 85-94, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30502321

RESUMO

BACKGROUND: Cancer associated fibroblasts (CAFs) are known to be crucial constituents of cancer microenvironment (CME) and play an important role in initiation, progression and metastasis of various types of cancer, such as oral cancer, pancreatic cancer, and gastric cancer. CAFs are usually derived from normal fibroblasts (NFs), but the mechanism of the transition in gastric cancer has not yet been fully elucidated. METHODS: qRT-PCR and western blot were employed to investigate differences of miR-141 and STAT4 expression respectively. The CAF-like features and wnt/ß-catenin pathway related proteins in NF or BMSC were assessed by qRT-PCR or western blot after treated with the conditioned medium from different indicated groups of gastric cancer cells. The invasion and migration ability of AGS cells after transfection were analyzed by Transwell assay and wound healing assay. Dual-luciferase report assay was employed to determine the direct binding of miR-141 to STAT4 3' UTR. RESULTS: For the first time, the present study found that STAT4 over-expression in gastric cancer cells induced NFs to obtain CAF-like features via activating wnt/ß-catenin pathway. Further gain-of-function and loss-of-function analysis revealed that miR-141 not only limited the migration and invasion of the gastric cancer cells, but also inhibited the transition of NFs and BMSC to CAFs. The luciferase assay indicated that miR-141 directly targeted the 3'-UTR predictive sequence of STAT4. CONCLUSION: Our data showed that miR-141 inhibited migration and invasion of gastric cancer cells and inhibited transition from NFs to CAFs via targeting STAT4/wnt/ß-catenin pathway.


Assuntos
Fibroblastos Associados a Câncer/patologia , Fibroblastos/patologia , MicroRNAs/genética , Fator de Transcrição STAT4/metabolismo , Neoplasias Gástricas/patologia , Diferenciação Celular/fisiologia , Movimento Celular/fisiologia , Humanos , Células-Tronco Mesenquimais/patologia , Invasividade Neoplásica/patologia , Microambiente Tumoral/fisiologia
19.
Int J Cancer ; 144(11): 2746-2761, 2019 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-30426475

RESUMO

miRNAs play a central role in the complex signaling network of cancer cells with the tumor microenvironment. Little is known on the origin of circulating miRNAs and their relationship with the tumor microenvironment in lung cancer. Here, we focused on the cellular source and relative contribution of different cell types to circulating miRNAs composing our risk classifier of lung cancer using in vitro/in vivo models and clinical samples. A cell-type specific expression pattern and topography of several miRNAs such as mir-145 in fibroblasts, mir-126 in endothelial cells, mir-133a in skeletal muscle cells was observed in normal and lung cancer tissues. Granulocytes and platelets are the major contributors of miRNAs release in blood. miRNAs modulation observed in plasma of lung cancer subjects was consistent with de-regulation of the same miRNAs observed during immunosuppressive conversion of immune cells. In particular, activated neutrophils showed a miRNA profile mirroring that observed in plasma of lung cancer subjects. Interestingly mir-320a secreted by neutrophils of high-risk heavy-smokers promoted an M2-like protumorigenic phenotype through downregulation of STAT4 when shuttled into macrophages. These findings suggest a multifactorial and nonepithelial cell-autonomous origin of circulating miRNAs associated with risk of lung cancer and that circulating miRNAs may act in paracrine signaling with causative role in lung carcinogenesis and immunosuppression.


Assuntos
MicroRNA Circulante/metabolismo , Neoplasias Pulmonares/imunologia , Macrófagos/imunologia , MicroRNAs/metabolismo , Evasão Tumoral/genética , Animais , Carcinogênese/imunologia , Linhagem Celular Tumoral , MicroRNA Circulante/sangue , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica/imunologia , Humanos , Pulmão/patologia , Neoplasias Pulmonares/sangue , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos SCID , MicroRNAs/sangue , Neutrófilos/imunologia , Neutrófilos/metabolismo , Fator de Transcrição STAT4/genética , Fator de Transcrição STAT4/metabolismo , Fumar Tabaco/sangue , Fumar Tabaco/imunologia , Ensaios Antitumorais Modelo de Xenoenxerto
20.
PLoS One ; 13(11): e0206459, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30395609

RESUMO

Mycobacterium tuberculosis (M.tb) contrives intracellular abode as a strategy to combat antibody onslaught. Additionally, to thrive against hostile ambiance inside host macrophages, the pathogen inhibits phago-lysosomal fusion. Finally, to further defy host cell offensives, M.tb opts for dormant phase, where it turns off or slows down most of its metabolic process as an added stratagem. While M.tb restrains most of its metabolic activities during dormancy, surprisingly latency-associated alpha-crystallin protein (Acr-1) is expressed most prominently during this phase. Interestingly, several previous studies described the potential of Acr-1 to induce the robust immuno-prophylactic response in the immunized host. It is intriguing to comprehend the apparent discrepancy that the microbe M.tb overexpresses a protein that has the potential to prime host immune system against the pathogen itself. Keeping this apparent ambiguity into consideration, it is imperative to unravel intricacies involved in the exploitation of Acr-1 by M.tb during its interaction with host immune cells. The present study suggests that Acr-1 exhibits diverse role in the maturation of macrophages (MΦs) and related immunological responses. The early encounter of bone marrow derived immune cells (pre-exposure during differentiation to MΦs) with Acr-1 (AcrMΦpre), results in hampering of their function. The pre-exposure of naïve MΦs with Acr-1 induces the expression of TIM-3 and IL-10. In contrast, exposure of fully differentiated MΦs to Acr-1 results in their down-modulation and induces the phosphorylation of STAT-1 and STAT-4 in host MΦs. Furthermore, Acr-1 mediated activation of MΦs results in the induction of Th1 and Th17 phenotype by activated T lymphocyte.


Assuntos
Proteínas de Bactérias/farmacologia , Interações Hospedeiro-Patógeno , Macrófagos/efeitos dos fármacos , Mycobacterium tuberculosis/metabolismo , Animais , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Citocinas/metabolismo , Humanos , Macrófagos/citologia , Macrófagos/metabolismo , Camundongos , Mycobacterium tuberculosis/fisiologia , Fenótipo , Fosforilação/efeitos dos fármacos , Fator de Transcrição STAT1/metabolismo , Fator de Transcrição STAT4/metabolismo , Linfócitos T/citologia , Linfócitos T/efeitos dos fármacos
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