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1.
Phytomedicine ; 124: 155280, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38183697

RESUMO

BACKGROUND: Polycystic ovary syndrome (PCOS) is the most common reproductive-endocrine condition in premenopausal women. Troxerutin, a common clinical anti-coagulant agent, was shown to work as a strong IL-22 boosting agent counteracting the hyperactivated gonadotrophin releasing hormone (GnRH) neurons and heightened GnRH release, the neuroendocrine origin of PCOS with unknown mechanism in rats. Exploring the off-label use of troxerutin medication for PCOS is thus sorely needed. METHODS: Serum IL-22 content and hypothalamic IL-22 protein were detected. Inflammatory factor levels in hypothalamo-pituitary were evaluated. Immunofluorescence staining was employed to determine the activation and M1/M2-prone polarization of microglia in arcuate hypothalamus and median eminence. RNA-sequencing and transcriptome analysis were applied to explore the potential driver of microglia M2-polarization in response to IL-22 bolstering effect. The function of microglial IL-22/IL-22R1/IRF3 system was further verified using in vivo knockdown of IL-22R1 and a potent IRF3 inhibitor in BV2 microglial cell lines in vitro. RESULTS: Troxerutin augmented serum IL-22 content, and its consequent spillover into the hypothalamus led to the direct activation of IL-22R1/IRF3 system on microglia, thereby promoted microglia M2 polarization in arcuate hypothalamus and median eminence, dampened hypothalamic neuroinflammation, inhibited hyperactive GnRH and rescued a breadth of PCOS-like traits in dihydrotestosterone (DHT) rats. The salutary effects of troxerutin treatment on hypothalamic neuroinflammation, microglial M1/2 polarization, GnRH secretion and numerous PCOS-like features were blocked by in vivo knockdown of IL-22R1. Moreover, evidence in vitro illustrated that IL-22 supplement to BV-2 microglia cell lines promoted M2 polarization, overproduction of anti-inflammatory marker and limitation of pro-inflammatory factors, whereas these IL-22 effects were blunted by geldanamycin, a potent IRF3 inhibitor. CONCLUSION: Here, the present study reported the potential off-label use of troxerutin medication, a common clinical anti-coagulant agent and an endogenous IL-22 enhancer, for multiple purposes in PCOS. The rational underlying the application of troxerutin as a therapeutic choice in PCOS derived from its activity as an IL-22 memetic agent targeting the neuro-endocrine origin of PCOS, and its promotive impact on microglia M2 polarization via activating microglial IL-22R1/IRF3 system in the arcuate hypothalamus and median eminence of DHT female rats.


Assuntos
Hidroxietilrutosídeo/análogos & derivados , Síndrome do Ovário Policístico , Receptores de Interleucina , Humanos , Ratos , Feminino , Animais , Síndrome do Ovário Policístico/induzido quimicamente , Síndrome do Ovário Policístico/tratamento farmacológico , Di-Hidrotestosterona/efeitos adversos , Di-Hidrotestosterona/metabolismo , Microglia , Doenças Neuroinflamatórias , Interleucina 22 , Hipotálamo/metabolismo , Hormônio Liberador de Gonadotropina/efeitos adversos , Hormônio Liberador de Gonadotropina/metabolismo , Fator Regulador 3 de Interferon/metabolismo
2.
JCI Insight ; 7(15)2022 08 08.
Artigo em Inglês | MEDLINE | ID: mdl-35763349

RESUMO

Current treatments fail to modify the underlying pathophysiology and disease progression of chronic obstructive pulmonary disease (COPD), necessitating alternative therapies. Here, we show that COPD subjects have increased IL-36γ and decreased IL-36 receptor antagonist (IL-36Ra) in bronchoalveolar and nasal fluid compared with control subjects. IL-36γ is derived from small airway epithelial cells (SAEC) and is further induced by a viral mimetic, whereas IL-36Ra is derived from macrophages. IL-36γ stimulates release of the neutrophil chemoattractants CXCL1 and CXCL8, as well as elastolytic matrix metalloproteinases (MMPs) from small airway fibroblasts (SAF). Proteases released from COPD neutrophils cleave and activate IL-36γ, thereby perpetuating IL-36 inflammation. Transfer of culture media from SAEC to SAF stimulated release of CXCL1, which was inhibited by exogenous IL-36Ra. The use of a therapeutic antibody that inhibits binding to the IL-36R attenuated IL-36γ-driven inflammation and cellular crosstalk. We have demonstrated a mechanism for the amplification and propagation of neutrophilic inflammation in COPD and have shown that blocking this cytokine family via a IL-36R neutralizing antibody could be a promising therapeutic strategy in the treatment of COPD.


Assuntos
Interleucina-1 , Doença Pulmonar Obstrutiva Crônica , Receptores de Interleucina/agonistas , Citocinas/metabolismo , Humanos , Inflamação/metabolismo , Interleucina-1/metabolismo , Interleucinas/metabolismo , Doença Pulmonar Obstrutiva Crônica/tratamento farmacológico
3.
Front Immunol ; 12: 778830, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34777396

RESUMO

Pathogenic inflammation and immuno-suppression are cardinal features of exhausted monocytes increasingly recognized in septic patients and murine models of sepsis. However, underlying mechanisms responsible for the generation of exhausted monocytes have not been addressed. In this report, we examined the generation of exhausted primary murine monocytes through prolonged and repetitive challenges with high dose bacterial endotoxin lipopolysaccharide (LPS). We demonstrated that repetitive LPS challenges skew monocytes into the classically exhausted Ly6Chi population, and deplete the homeostatic non-classical Ly6Clo population, reminiscent of monocyte exhaustion in septic patients. scRNAseq analyses confirmed the expansion of Ly6Chi monocyte cluster, with elevation of pathogenic inflammatory genes previously observed in human septic patients. Furthermore, we identified CD38 as an inflammatory mediator of exhausted monocytes, associated with a drastic depletion of cellular NAD+; elevation of ROS; and compromise of mitochondria respiration, representative of septic monocytes. Mechanistically, we revealed that STAT1 is robustly elevated and sustained in LPS-exhausted monocytes, dependent upon the TRAM adaptor of the TLR4 pathway. TRAM deficient monocytes are largely resistant to LPS-mediated exhaustion, and retain the non-classical homeostatic features. Together, our current study addresses an important yet less-examined area of monocyte exhaustion, by providing phenotypic and mechanistic insights regarding the generation of exhausted monocytes.


Assuntos
Memória Imunológica , Inflamação/imunologia , Monócitos/imunologia , Sepse/imunologia , ADP-Ribosil Ciclase 1/genética , ADP-Ribosil Ciclase 1/metabolismo , Animais , Antígenos Ly/genética , Antígenos Ly/metabolismo , Antígeno B7-2/genética , Antígeno B7-2/metabolismo , Células Cultivadas , Memória Imunológica/efeitos dos fármacos , Inflamação/genética , Inflamação/metabolismo , Fator 4 Semelhante a Kruppel/metabolismo , Lipopolissacarídeos/farmacologia , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , Camundongos Endogâmicos C57BL , Camundongos Knockout , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Fenótipo , Receptores de Interleucina/genética , Receptores de Interleucina/metabolismo , Fator de Transcrição STAT1/metabolismo , Sepse/genética , Sepse/metabolismo , Transdução de Sinais , Receptor 4 Toll-Like/metabolismo
4.
Int J Biol Macromol ; 185: 696-707, 2021 Aug 31.
Artigo em Inglês | MEDLINE | ID: mdl-34174316

RESUMO

The inspection of variations in the proteomic aspects conspire the biomarker discovery in diagnostics of peculiar diseases. Recent developments in high-throughput proteomic techniques have provided leverage in the discovery of biomarkers during the etiology of various diseases. We identified potential biomarkers by utilizing proteomics, bioinformatics and gene expression studies. Meticulous assessment of collagen and hydroxyproline levels along with the glycogen and protein carbonyl levels exhibited deterioration in the N' - Nitrosodiethylamine (NDEA) administered rat livers and subsequent salubrious effect of pomegranate juice. The immunohistochemical inspection of iNOS and nitrite estimation indicated the peccant fibrotic alterations. 2D proteome profiling and MALDI-TOF MS/MS furthered the significant biomarkers to be analyzed for the gene ontology by PANTHER, cluster analysis by DAVID and network simulation by STRING 10.0. Several genes found relevant after MALDI analysis were evaluated by real-time PCR (RTPCR). Our data revealed CYP2b15, HSP70, TRFE, HPT, Il1rl2, Ric8a, Krt18, Hsp90b1 and iNOS as novel biomarkers for the mechanism of pomegranate against liver fibrosis. It can be inferred that NDEA-induced liver fibrosis actuates various biological pathways by the identified biomarkers and pomegranate juice modifies them.


Assuntos
Biomarcadores/metabolismo , Dimetilnitrosamina/efeitos adversos , Cirrose Hepática/tratamento farmacológico , Extratos Vegetais/administração & dosagem , Punica granatum/química , Animais , Suplementos Nutricionais , Modelos Animais de Doenças , Sucos de Frutas e Vegetais , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Fatores de Troca do Nucleotídeo Guanina/genética , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Queratina-18/genética , Queratina-18/metabolismo , Cirrose Hepática/induzido quimicamente , Cirrose Hepática/metabolismo , Masculino , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Extratos Vegetais/farmacologia , Proteômica , Ratos , Receptores de Interleucina/genética , Receptores de Interleucina/metabolismo , Espectrometria de Massas em Tandem
5.
Acupunct Med ; 39(6): 663-672, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33715422

RESUMO

OBJECTIVE: Interleukin (IL)-17, as a T-helper 17 cell (Th17) cytokine, plays a key role in chronic obstructive pulmonary disease (COPD) pathophysiology including chronic inflammation and airway obstruction, which lead to decreased pulmonary function. The aim of this study was to investigate the effect of acupuncture on IL-17, its receptor (IL-17R) and the mitogen-activated protein kinase (MAPK) signaling pathway, in a rat model of COPD. METHODS: The COPD model was induced in Sprague Dawley rats by exposure to cigarette smoke for 12 weeks. The model rats were treated with electroacupuncture (EA) at BL13 and ST36. The lung function and histology of the rats were observed. IL-17, tumor necrosis factor (TNF)-α, and IL-10 were detected by enzyme-linked immunosorbent assay (ELISA) in bronchoalveolar lavage fluid (BALF) and in plasma. The leukocytes and macrophages in the BALF were counted. The expression levels of IL-17R were assayed in lung tissue by real-time polymerase chain reaction (PCR), western blotting, and immunohistochemistry. MAPK signaling pathway molecules including c-Jun N-terminal kinase (JNK), extracellular signal-regulated kinase (ERK)1/2 and p38, and their phosphorylated forms, were observed in the lung by western blotting. RESULTS: Compared with the control group rats, lung function decreased and there was a severe inflammatory infiltration of the pulmonary parenchyma in the COPD rats. EA effectively improved lung function and alleviated the inflammatory infiltration in the lungs of COPD rats. EA also reversed the elevated total leukocyte and macrophage counts, the high levels of IL-17 and TNF-α, and the low IL-10 content in COPD rats. Meanwhile, EA downregulated the increased mRNA and protein expression of IL-17R, and significantly inhibited the elevated levels of phosphorylated JNK, ERK1/2, and p38 in the lungs of COPD rats. CONCLUSION: Our results suggest that the protective effects of acupuncture therapy on the lungs of COPD rats are likely related to inhibition of IL-17/IL-17R and the post-receptor MAPK signaling pathways.


Assuntos
Eletroacupuntura , Sistema de Sinalização das MAP Quinases , Doença Pulmonar Obstrutiva Crônica/terapia , Receptores de Interleucina/sangue , Animais , Líquido da Lavagem Broncoalveolar/química , Modelos Animais de Doenças , Humanos , Interleucina-10/sangue , Interleucina-10/líquido cefalorraquidiano , Interleucina-17/sangue , Interleucina-17/líquido cefalorraquidiano , Pulmão/metabolismo , Masculino , Doença Pulmonar Obstrutiva Crônica/sangue , Ratos , Ratos Sprague-Dawley , Fator de Necrose Tumoral alfa/sangue , Fator de Necrose Tumoral alfa/líquido cefalorraquidiano
6.
Eur J Immunol ; 51(1): 191-196, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-32648940

RESUMO

Interleukin-31 (IL-31) is a Th2 cell-derived cytokine that has been closely linked to pruritic skin inflammation. More recently, enhanced IL-31 serum levels have also been observed in patients with allergic rhinitis and allergic asthma. Therefore, the main aim of this study was to unravel the contribution of IL-31 to allergen-induced lung inflammation. We analyzed lung inflammation in response to the timothy grass (Phleum pratense) pollen allergen Phl p 5 in C57BL/6 wild-type (wt) mice, IL-31 transgenic (IL-31tg) mice, and IL-31 receptor alpha-deficient animals (IL-31RA-/- ). IL-31 and IL-31RA levels were monitored by qRT-PCR. Cellular infiltrate in bronchoalveolar lavage fluid (BALF) and lung tissue inflammation, mucus production as well as epithelial thickness were measured by flow cytometry and histomorphology. While allergen challenge induced IL-31RA expression in lung tissue of wt and IL-31tg mice, high IL-31 expression was exclusively observed in lung tissue of IL-31tg mice. Upon Phl p 5 challenge, IL-31tg mice showed reduced numbers of leukocytes and eosinophils in BALF and lung tissue as well as diminished mucin expression and less pronounced epithelial thickening compared to IL-31RA-/- or wt animals. These findings suggest that the IL-31/IL-31RA axis may regulate local, allergen-induced inflammation in the lungs.


Assuntos
Alérgenos/efeitos adversos , Alérgenos/imunologia , Interleucinas/imunologia , Proteínas de Plantas/efeitos adversos , Proteínas de Plantas/imunologia , Pneumonia/imunologia , Animais , Asma/etiologia , Asma/imunologia , Asma/prevenção & controle , Líquido da Lavagem Broncoalveolar/imunologia , Modelos Animais de Doenças , Eosinófilos/imunologia , Feminino , Interleucinas/genética , Leucócitos/imunologia , Pulmão/imunologia , Pulmão/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Camundongos Transgênicos , Phleum/efeitos adversos , Phleum/imunologia , Pneumonia/etiologia , Pneumonia/prevenção & controle , Pólen/efeitos adversos , Pólen/imunologia , Receptores de Interleucina/deficiência , Receptores de Interleucina/genética , Receptores de Interleucina/imunologia
7.
Expert Opin Biol Ther ; 21(2): 271-277, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33216643

RESUMO

Background: The susceptibility of patients with chronic plaque psoriasis and the risks or benefits related to the use of biological therapies for COVID-19 are unknown. Few data about prevalence, clinical course and outcomes of COVID-19 among psoriatic patients were reported. The aims of this study were 1) to assess the prevalence and severity of COVID-19 in psoriatic patients treated with biologic agents during the first phase of the emergency (22 February to 22 April 2020) in Italy, and 2) to report the clinical outcomes of patients who have been exposed to individuals with confirmed SARS-CoV-2 infection. Methods: Patients with moderate-to-severe chronic plaque psoriasis, aged ≥18 years and undergoing treatment with biologic agents as of 22 February 2020, were eligible to be included in PSO-BIO-COVID study. Demographic and clinical characteristics of patients using any biologic for psoriasis treatment between 22 February and 22 April 2020 were registered. Results: A total of 12,807 psoriatic patients were included in the PSO-BIO-COVID study. In this cohort 26 patients (0.2%) had a swab confirmation of SARS-CoV-2 infection. Eleven patients required hospitalization and two died. Conclusion: The incidence of COVID-19 observed in our cohort of psoriatic patients (0.2%) is similar to that seen in the general population (0.31%) in Italy. However, the course of the disease was mild in most patients. Biological therapies may likely lessen 'cytokine storm' of COVID-19, which sometimes lead to multiple organ failure, ARDS, and death.


Assuntos
Produtos Biológicos/uso terapêutico , Terapia Biológica/métodos , COVID-19/epidemiologia , Psoríase/tratamento farmacológico , Adulto , Idoso , Idoso de 80 Anos ou mais , Produtos Biológicos/farmacologia , COVID-19/diagnóstico , Doença Crônica , Estudos de Coortes , Feminino , Humanos , Incidência , Interleucina-17/antagonistas & inibidores , Itália/epidemiologia , Masculino , Pessoa de Meia-Idade , Pandemias , Psoríase/diagnóstico , Psoríase/epidemiologia , Receptores de Interleucina/antagonistas & inibidores , Medição de Risco/métodos , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Adulto Jovem
8.
Microb Pathog ; 138: 103826, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31676364

RESUMO

Tangeretin, a polymethoxylated flavonoid is abundant in citrus fruits, which has been reported to inhibit inflammation by inhibiting NF-κB activation and proinflammatory cytokines. Notch blockage inhibits Th17 cells response that are involved in the development of acute lung injury (ALI). This study investigated the protective effects of tangeretin on LPS-induced ALI in mice. Male C57BL/6 mice were treated with phosphate-buffered saline (PBS), lipopolysaccharide (LPS), LPS and tangeretin, or LPS and N-[N-(3,5-difluorophenacetyl)-l-alanyl]-S-phenylglycine t-butyl ester (DAPT, a Notch signaling inhibitor), which were harvested at 48 h after challenged by LPS. CD4+ T cells were treated with tangeretin or DAPT and harvested after 72 h. Tangeretin notably attenuated pathological changes and decreased the wet to dry weight ratio of the mouse lungs. The total cell and neutrophil counts, tumor necrosis factor (TNF)-α in bronchoalveolar lavage fluid (BALF), myeloperoxidase activity of lung tissue were markedly reduced by tangeretin. The percentage of CD4+IL-17 + T cells in the lungs and the concentration of interleukin (IL)-17 and IL-22 in BALF were significantly down-regulated by tangeretin. As with the positive control (DAPT), tangeretin inhibited the activity of the Notch signaling pathway accompanied with the down-regulation of acid-related orphan receptor gamma t and IL-23 receptor expression. This study demonstrated that tangeretin protects against LPS-induced ALI by suppressing Th17 response at least partially, through a Notch-dependent mechanism.


Assuntos
Lesão Pulmonar Aguda/etiologia , Lesão Pulmonar Aguda/metabolismo , Flavonas/farmacologia , Receptores Notch/metabolismo , Transdução de Sinais , Células Th17/efeitos dos fármacos , Células Th17/metabolismo , Lesão Pulmonar Aguda/patologia , Animais , Biomarcadores , Modelos Animais de Doenças , Suscetibilidade a Doenças , Flavonas/química , Citometria de Fluxo , Lipopolissacarídeos/efeitos adversos , Masculino , Camundongos , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/genética , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/metabolismo , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Receptores de Interleucina/metabolismo , Células Th17/imunologia
9.
Int J Mol Sci ; 20(5)2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30823645

RESUMO

Maternal diet modifies epigenetic programming in offspring, a potentially critical factor in the immune dysregulation of modern societies. We previously found that prenatal fish oil supplementation affects neonatal T-cell histone acetylation of genes implicated in adaptive immunity including PRKCZ, IL13, and TBX21. In this study, we measured H3 and H4 histone acetylation levels by chromatin immunoprecipitation in 173 term placentas collected in the prospective birth cohort, ALADDIN, in which information on lifestyle and diet is thoroughly recorded. In anthroposophic families, regular olive oil usage during pregnancy was associated with increased H3 acetylation at FOXP3 (p = 0.004), IL10RA (p = 0.008), and IL7R (p = 0.007) promoters, which remained significant after adjustment by offspring gender. Furthermore, maternal fish consumption was associated with increased H4 acetylation at the CD14 gene in placentas of female offspring (p = 0.009). In conclusion, prenatal olive oil intake can affect placental histone acetylation in immune regulatory genes, confirming previously observed pro-acetylation effects of olive oil polyphenols. The association with fish consumption may implicate ω-3 polyunsaturated fatty acids present in fish oil. Altered histone acetylation in placentas from mothers who regularly include fish or olive oil in their diets could influence immune priming in the newborn.


Assuntos
Óleos de Peixe/farmacologia , Histonas/metabolismo , Fenômenos Fisiológicos da Nutrição Materna , Azeite de Oliva/farmacologia , Placenta/metabolismo , Processamento de Proteína Pós-Traducional , Acetilação , Feminino , Óleos de Peixe/administração & dosagem , Óleos de Peixe/metabolismo , Produtos Pesqueiros , Humanos , Imunidade Inata/genética , Interleucina-13/genética , Interleucina-13/metabolismo , Receptores de Lipopolissacarídeos/genética , Receptores de Lipopolissacarídeos/metabolismo , Azeite de Oliva/administração & dosagem , Placenta/efeitos dos fármacos , Gravidez , Proteína Quinase C/genética , Proteína Quinase C/metabolismo , Receptores de Interleucina/genética , Receptores de Interleucina/metabolismo , Proteínas com Domínio T/genética , Proteínas com Domínio T/metabolismo
10.
Planta Med ; 84(15): 1110-1117, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29763944

RESUMO

Baicalin is the main flavonoid from the roots of an important medicinal plant, Scutellaria baicalensis, which shows a variety biological activities. Psoriasis is a chronic immune-mediated inflammatory disease that affects the skin. The unmet need of psoriasis is that many patients do not respond adequately to available clinical treatment. In this study, we found that baicalin showed inhibited dermal inflammation in a murine model of psoriasis via topical application of imiquimod. After a 5-day topical imiquimod application, baicalin or the control vehicle cream was to applied to the lesions of BALB/c mice for a further 4 days. The erythema, scaling, and thickness of the epidermal layer significantly improved in the baicalin-treated mice. The levels of interleukin-17A, interleukin-22, interleukin-23, and tumor necrosis factor in the skin significantly decreased after baicalin treatment. Baicalin also inhibited imiquimod-induced interleukin-17A production in skin draining lymph node cells. The infiltration of γδ T cells into the skin lesions induced by imiquimod was also suppressed after baicalin treatment. These results suggest that baicalin inhibited skin inflammation through the inhibition of the interleukin-17/interleukin-23 axis in a murine model of psoriasis.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Citocinas/metabolismo , Toxidermias/tratamento farmacológico , Flavonoides/farmacologia , Psoríase/tratamento farmacológico , Aminoquinolinas/efeitos adversos , Animais , Anti-Inflamatórios não Esteroides/química , Modelos Animais de Doenças , Toxidermias/patologia , Feminino , Flavonoides/química , Humanos , Imiquimode , Interleucina-17/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Psoríase/patologia , Receptores de Interleucina/metabolismo , Pele/patologia
11.
PLoS One ; 12(11): e0188391, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-29155882

RESUMO

Retinoic acid receptor-related-orphan-receptor-C (RORγt) is the key transcription factor that is driving the differentiation of IL-17 producing T-helper 17 (Th17) cells that are implicated in the pathology of various autoimmune and inflammatory diseases. Based on the importance of RORγt in promoting Th17-driven pathology, there is considerable interest to develop low-molecular-weight compounds with the aim of inhibiting the transcriptional activity of this nuclear hormone receptor. In this article, we describe the in vitro and in vivo pharmacology of a potent and selective small-molecular-weight RORγt inverse agonist. The compound binds to the ligand binding domain (LBD) of RORγt leading to displacement of a co-activator peptide. We show for the first time that a RORγt inverse agonist down-regulates permissive histone H3 acetylation and methylation at the IL17A and IL23R promoter regions, thereby providing insight into the transcriptional inhibition of RORγt-dependent genes. Consistent with this, the compound effectively reduced IL-17A production by polarized human T-cells and γδT-cells and attenuated transcription of RORγt target genes. The inhibitor showed good in vivo efficacy in an antigen-induced arthritis model in rats and reduced the frequencies of IL-17A producing cells in ex vivo recall assays. In summary, we demonstrate that inhibiting RORγt by a low-molecular-weight inhibitor results in efficient and selective blockade of the pro-inflammatory Th17/IL-17A pathway making it an attractive target for Th17-mediated disorders.


Assuntos
Artrite Experimental/tratamento farmacológico , Imidazóis/farmacologia , Interleucina-17/antagonistas & inibidores , Linfócitos Intraepiteliais/efeitos dos fármacos , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/antagonistas & inibidores , Piridinas/farmacologia , Pirimidinas/farmacologia , Células Th17/efeitos dos fármacos , Animais , Artrite Experimental/genética , Artrite Experimental/imunologia , Artrite Experimental/patologia , Linhagem Celular Tumoral , Feminino , Regulação da Expressão Gênica , Células HEK293 , Humanos , Imidazóis/síntese química , Interleucina-17/genética , Interleucina-17/imunologia , Linfócitos Intraepiteliais/imunologia , Linfócitos Intraepiteliais/patologia , Cinética , Masculino , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/genética , Membro 3 do Grupo F da Subfamília 1 de Receptores Nucleares/imunologia , Regiões Promotoras Genéticas , Ligação Proteica , Piridinas/síntese química , Pirimidinas/síntese química , Ratos , Ratos Endogâmicos Lew , Receptores de Antígenos de Linfócitos T gama-delta/genética , Receptores de Antígenos de Linfócitos T gama-delta/imunologia , Receptores de Interleucina/genética , Receptores de Interleucina/imunologia , Transdução de Sinais , Células Th17/imunologia , Células Th17/patologia
12.
Proc Natl Acad Sci U S A ; 114(38): 10190-10195, 2017 09 19.
Artigo em Inglês | MEDLINE | ID: mdl-28874534

RESUMO

Dysregulated Foxp3+ Treg functions result in uncontrolled immune activation and autoimmunity. Therefore, identifying cellular factors modulating Treg functions is an area of great importance. Here, using Treg-specific Il27ra-/- mice, we report that IL-27 signaling in Foxp3+ Tregs is essential for Tregs to control autoimmune inflammation in the central nervous system (CNS). Following experimental autoimmune encephalomyelitis (EAE) induction, Treg-specific Il27ra-/- mice develop more severe EAE. Consistent with the severe disease, the numbers of IFNγ- and IL-17-producing CD4 T cells infiltrating the CNS tissues are greater in these mice. Treg accumulation in the inflamed CNS tissues is not affected by the lack of IL-27 signaling in Tregs, suggesting a functional defect of Il27ra-/- Tregs. IL-10 production by conventional CD4 T cells and their CNS accumulation are rather elevated in Treg-specific Il27ra-/- mice. Analysis with Treg fate-mapping reporter mice further demonstrates that IL-27 signaling in Tregs may control stability of Foxp3 expression. Finally, systemic administration of recombinant IL-27 in Treg-specific Il27ra-/- mice fails to ameliorate the disease even in the presence of IL-27-responsive conventional CD4 T cells. These findings uncover a previously unknown role of IL-27 in regulating Treg function to control autoimmune inflammation.


Assuntos
Doenças Autoimunes/imunologia , Encefalomielite/imunologia , Receptores de Citocinas/metabolismo , Linfócitos T Reguladores/metabolismo , Animais , Doenças Autoimunes/tratamento farmacológico , Sistema Nervoso Central/imunologia , Avaliação Pré-Clínica de Medicamentos , Encefalomielite/tratamento farmacológico , Fatores de Transcrição Forkhead/metabolismo , Interleucinas/metabolismo , Interleucinas/uso terapêutico , Camundongos Transgênicos , Receptores de Citocinas/genética , Receptores de Interleucina
13.
Int J Dermatol ; 55(7): e380-5, 2016 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-26748443

RESUMO

BACKGROUND: Effectiveness of ultraviolet (UV)A1 in flares of atopic dermatitis (AD) is thought to influence the expression of cytokines involved in its pathogenesis. The aim of the study was to investigate whether mRNA expression of human ß defensin-1 (hßD-1) correlates with that of interleukin (IL)-4, IL-10, and IL-31 in skin lesions in AD before and after UVA1 phototherapy, to determine whether UVA1 decreases the expression of the aforementioned mediators, and to confirm whether changes in mRNA expression correspond with the clinical efficacy of UVA1. METHODS: Twenty-five patients with AD underwent medium-dose UVA1 phototherapy. Before and after UVA1, biopsies from acute skin lesions were studied using reverse transcription and real-time polymerase chain reaction. RESULTS: Levels of mRNA hßD-1 correlated with those of IL-10 and IL-31, levels of IL-4 mRNA correlated with those of IL-10 and IL-31, and IL-10 expression correlated with that of IL-31, both before and after UVA1. Phototherapy with UVA1 improved SCORing of Atopic Dermatitis (SCORAD) values, decreased pruritus, and increased expression of IL-4. After UVA1, no difference was found in the mRNA expression of other molecules. The SCORAD index did not correlate with the expression of any examined mRNA either before or after UVA1. CONCLUSIONS: hßD-1, IL-4, IL-10, and IL-31 are expressed in acute skin lesions in AD, and their levels correlate with each other. UVA1 improves SCORAD and pruritus and increases the expression of IL-4 without direct effect on other molecules.


Assuntos
Dermatite Atópica/genética , Dermatite Atópica/radioterapia , Interleucinas/genética , RNA Mensageiro/metabolismo , Terapia Ultravioleta , beta-Defensinas/genética , Adolescente , Adulto , Dermatite Atópica/complicações , Feminino , Expressão Gênica/efeitos da radiação , Humanos , Interleucina-10/genética , Interleucina-4/genética , Masculino , Pessoa de Meia-Idade , Prurido/etiologia , Prurido/radioterapia , Dosagem Radioterapêutica , Receptores de Interleucina/genética , Índice de Gravidade de Doença , Adulto Jovem
14.
J Immunol ; 195(7): 3001-10, 2015 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-26324771

RESUMO

IL-23 is the key cytokine that induces the expansion of Th17 cells. It is composed of p19 and p40 subunits of IL-12. The p40 subunit binds competitively to the receptor of IL-23 and blocks its activity. Our aim was to assess the preventive and therapeutic effect of the IL-12p40 homodimer (p40)2 subunit in autoimmune arthritis animal models. In the current study, using IL-1R antagonist-knockout mice and a collagen-induced arthritis model, we investigated the suppressive effect of (p40)2 on inflammatory arthritis. We demonstrated that the recombinant adenovirus-expressing mouse (p40)2 model prevented the development of arthritis when given before the onset of arthritis. It also decreased the arthritis index and joint erosions in the mouse model if transferred after arthritis was established. (p40)2 inhibited the production of inflammatory cytokines and Ag-specific T cell proliferation. It also induced CD4(+)CD25(+)Foxp3 regulatory T (Treg) cells in vitro and in vivo, whereas the generation of retinoic acid receptor-related organ receptor γt and Th17 cells was suppressed. The induction of Treg cells and the suppression of Th17 cells were mediated via activated STAT5 and suppressed STAT3. Our data suggest that (p40)2 suppressed inflammatory arthritis successfully. This could be a useful therapeutic approach in autoimmune arthritis to regulate the Th17/Treg balance and IL-23 signaling.


Assuntos
Artrite Experimental/prevenção & controle , Subunidade p40 da Interleucina-12/farmacologia , Subunidade p19 da Interleucina-23/imunologia , Linfócitos T Reguladores/imunologia , Células Th17/imunologia , Animais , Artrite Experimental/tratamento farmacológico , Artrite Experimental/imunologia , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Colágeno/imunologia , Citocinas/biossíntese , Subunidade p40 da Interleucina-12/imunologia , Interleucina-17/biossíntese , Ativação Linfocitária/imunologia , Masculino , Camundongos , Camundongos Endogâmicos DBA , Camundongos Knockout , Membro 1 do Grupo F da Subfamília 1 de Receptores Nucleares/biossíntese , Multimerização Proteica , Receptores de Interleucina/imunologia , Receptores Tipo I de Interleucina-1/genética , Fator de Transcrição STAT3/metabolismo , Fator de Transcrição STAT5/metabolismo , Transdução de Sinais/imunologia
15.
PLoS One ; 10(6): e0129576, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26067287

RESUMO

Although acupuncture is widely used to manage pain, it remains highly controversial, largely due to the lack of a clear mechanism for its benefits. Here, we investigated the role of IL-33, a novel interleukin (IL)-1 family member, and its receptor ST2 in the analgesic effects of electroacupuncture (EA) on formalin-induced inflammatory pain. The results showed that 1) EA stimulation of ipsilateral Zusanli (ST 36) and Yanglingquan (GB 34) acupoints for 30 min remarkably suppressed the two phases of formalin-induced spontaneous pain; 2) subcutaneous or intrathecal administration of recombinant IL-33 (rIL-33) significantly inhibited the analgesic effect of EA, whereas the ST2 antibody potentiated EA analgesia in formalin mice; 3) EA treatment decreased the up-regulation of IL-33 and ST2 protein following formalin injection; and 4) the suppression of the formalin-induced expression of spinal phosphorylated ERK and JNK induced by EA treatment was significantly attenuated following subcutaneous rIL-33 delivery, and was further decreased by the ST2 antibody. These data suggest that EA alleviates formalin-induced inflammatory pain, at least partially, by inhibiting of spinal IL-33/ST2 signaling and the downstream ERK and JNK pathways.


Assuntos
Eletroacupuntura , Interleucina-33/metabolismo , Sistema de Sinalização das MAP Quinases , Neuralgia/terapia , Animais , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Formaldeído/toxicidade , Proteína 1 Semelhante a Receptor de Interleucina-1 , Interleucina-33/uso terapêutico , MAP Quinase Quinase 4/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Neuralgia/tratamento farmacológico , Neuralgia/etiologia , Neuralgia/metabolismo , Receptores de Interleucina/metabolismo
16.
Expert Rev Proteomics ; 12(2): 159-69, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25711416

RESUMO

IL-23 is an important therapeutic target for the treatment of inflammatory diseases. Adnectins are targeted protein therapeutics that are derived from domain III of human fibronectin and have a similar protein scaffold to antibodies. Adnectin 2 was found to bind to IL-23 and compete with the IL-23/IL-23R interaction, posing a potential protein therapeutic. Hydrogen/deuterium exchange mass spectrometry and computational methods were applied to probe the binding interactions between IL-23 and Adnectin 2 and to determine the correlation between the two orthogonal methods. This review summarizes the current structural knowledge about IL-23 and focuses on the applicability of hydrogen/deuterium exchange mass spectrometry to investigate the higher order structure of proteins, which plays an important role in the discovery of new and improved biotherapeutics.


Assuntos
Terapia Biológica , Deutério/química , Hidrogênio/química , Interleucina-23/química , Biologia Computacional , Humanos , Interleucina-23/metabolismo , Espectrometria de Massas/métodos , Ligação Proteica , Conformação Proteica , Receptores de Interleucina/química
17.
J Immunol ; 194(1): 316-24, 2015 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-25452564

RESUMO

IL-23 has been well studied in the context of T cell differentiation; however, its role in the differentiation of myeloid progenitors is less clear. In this paper, we describe a novel role of IL-23 in myeloid cell differentiation. Specifically, we have identified that in human PBMCs, IL-23 induces the expression of MDL-1, a PU.1 transcriptional target during myeloid differentiation, which orchestrates osteoclast differentiation through activation of DNAX activating protein of 12 kDa and its ITAMs. The molecular events that lead to the differentiation of human macrophages to terminally differentiated osteoclasts are dependent on spleen tyrosine kinase and phospholipase Cγ2 phosphorylation for the induction of intracellular calcium flux and the subsequent activation of master regulator osteoclast transcription factor NFATc1. IL-23-elicited osteoclastogenesis is independent of the receptor activator of NF-κB ligand pathway and uses a unique myeloid DNAX activating protein of 12 kDa-associated lectin-1(+)/DNAX activating protein of 12 kDa(+) cell subset. Our data define a novel pathway that is used by IL-23 in myeloid cells and identify a major mechanism for the stimulation of osteoclastogenesis in inflammatory arthritis.


Assuntos
Artrite/imunologia , Interleucina-23/metabolismo , Macrófagos/citologia , Células Progenitoras Mieloides/citologia , Osteoclastos/metabolismo , Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Artrite/metabolismo , Cálcio/metabolismo , Diferenciação Celular , Células Cultivadas , Ativação Enzimática , Humanos , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Lectinas Tipo C/biossíntese , Ativação de Macrófagos/imunologia , Macrófagos/imunologia , Proteínas de Membrana/metabolismo , Complexos Multiproteicos/biossíntese , Fatores de Transcrição NFATC/biossíntese , Fatores de Transcrição NFATC/metabolismo , Osteoclastos/citologia , Fosfolipase C gama/metabolismo , Fosforilação , Estrutura Quaternária de Proteína , Proteínas Tirosina Quinases/metabolismo , Ligante RANK/metabolismo , Receptores de Superfície Celular/biossíntese , Receptores de Interleucina/metabolismo , Transdução de Sinais , Quinase Syk
18.
Proc Natl Acad Sci U S A ; 111(28): 10281-6, 2014 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-24982172

RESUMO

Mast cells (MC) are potent innate immune cells that accumulate in chronically inflamed tissues. MC express the IL-33 receptor IL-1 receptor-related protein ST2 at high level, and this IL-1 family cytokine both activates MC directly and primes them to respond to other proinflammatory signals. Whether IL-33 and ST2 play a role in MC survival remains to be defined. In skin-derived human MC, we found that IL-33 attenuated MC apoptosis without altering proliferation, an effect mediated principally through the antiapoptotic molecule B-cell lymphoma-X large (BCLXL). Murine MC demonstrated a similar mechanism, dependent entirely on ST2. In line with these observations, St2(-/-) mice exhibited reduced numbers of tissue MC in inflamed arthritic joints, in helminth-infected intestine, and in normal peritoneum. To confirm an MC-intrinsic role for ST2 in vivo, we performed peritoneal transfer of WT and St2(-/-) MC. In St2(-/-) hosts treated with IL-33 and in WT hosts subjected to thioglycollate peritonitis, WT MC displayed a clear survival advantage over coengrafted St2(-/-) MC. IL-33 blockade specifically attenuated this survival advantage, confirming IL-33 as the relevant ST2 ligand mediating MC survival in vivo. Together, these data reveal a cell-intrinsic role for the IL-33/ST2 axis in the regulation of apoptosis in MC, identifying thereby a previously unappreciated pathway supporting expansion of the MC population with inflammation.


Assuntos
Interleucinas/metabolismo , Mastócitos/metabolismo , Receptores de Superfície Celular/metabolismo , Receptores de Interleucina/metabolismo , Proteína bcl-X/metabolismo , Animais , Artrite/genética , Artrite/imunologia , Artrite/metabolismo , Artrite/patologia , Sobrevivência Celular/genética , Sobrevivência Celular/imunologia , Helmintíase/genética , Helmintíase/imunologia , Helmintíase/metabolismo , Helmintíase/patologia , Humanos , Inflamação/imunologia , Inflamação/metabolismo , Inflamação/patologia , Proteína 1 Semelhante a Receptor de Interleucina-1 , Interleucina-33 , Interleucinas/genética , Interleucinas/imunologia , Mucosa Intestinal/metabolismo , Intestinos/imunologia , Intestinos/parasitologia , Articulações/imunologia , Articulações/metabolismo , Articulações/patologia , Mastócitos/imunologia , Mastócitos/patologia , Camundongos , Camundongos Knockout , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/imunologia , Receptores de Interleucina/genética , Receptores de Interleucina/imunologia , Proteína bcl-X/genética , Proteína bcl-X/imunologia
19.
Expert Opin Ther Targets ; 18(5): 513-25, 2014 May.
Artigo em Inglês | MEDLINE | ID: mdl-24568095

RESUMO

INTRODUCTION: Psoriasis is a chronic inflammatory skin disorder determined by the activation of several immune cells and resident tissue cells. Various cytokines mediate inflammatory signals, including IL-23, which is an important factor involved in the differentiation of T helper (Th17) cells. AREAS COVERED: Increasing evidence suggests that IL-23 is a central cytokine to the pathogenesis of psoriasis. An overview on both experimental and human data will be reported in order to support the hypothesis of a key pathogenic role of IL-23/Th17 axis. EXPERT OPINION: Targeting IL-23 might be a more selective, valid and effective therapeutic approach, which, potentially, may show important advantages in terms of long-term efficacy and safety in the treatment of psoriasis.


Assuntos
Anticorpos Monoclonais/uso terapêutico , Interleucina-23/fisiologia , Terapia de Alvo Molecular , Psoríase/fisiopatologia , Animais , Células Apresentadoras de Antígenos/metabolismo , Infecções Bacterianas/imunologia , Diferenciação Celular , Ensaios Clínicos como Assunto , Citocinas/fisiologia , Avaliação Pré-Clínica de Medicamentos , Predisposição Genética para Doença , Humanos , Subunidade p40 da Interleucina-12/antagonistas & inibidores , Subunidade p40 da Interleucina-12/genética , Interleucina-23/antagonistas & inibidores , Interleucina-23/genética , Subunidade p19 da Interleucina-23/antagonistas & inibidores , Subunidade p19 da Interleucina-23/genética , Subunidade p19 da Interleucina-23/fisiologia , Queratinócitos/metabolismo , Camundongos , Camundongos Knockout , Psoríase/genética , Psoríase/imunologia , Receptores de Interleucina/antagonistas & inibidores , Receptores de Interleucina/fisiologia , Transdução de Sinais , Subpopulações de Linfócitos T/imunologia , Células Th17/imunologia
20.
Proteins ; 82(6): 975-89, 2014 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-24549990

RESUMO

Engineered combinatorial libraries derived from small protein scaffolds represent a powerful tool for generating novel binders with high affinity, required specificity and designed inhibitory function. This work was aimed to generate a collection of recombinant binders of human interleukin-23 receptor (IL-23R), which is a key element of proinflammatory IL-23-mediated signaling. A library of variants derived from the three-helix bundle scaffold of the albumin-binding domain (ABD) of streptococcal protein G and ribosome display were used to select for high-affinity binders of recombinant extracellular IL-23R. A collection of 34 IL-23R-binding proteins (called REX binders), corresponding to 18 different sequence variants, was used to identify a group of ligands that inhibited binding of the recombinant p19 subunit of IL-23, or the biologically active human IL-23 cytokine, to the recombinant IL-23R or soluble IL-23R-IgG chimera. The strongest competitors for IL-23R binding in ELISA were confirmed to recognize human IL-23R-IgG in surface plasmon resonance experiments, estimating the binding affinity in the sub- to nanomolar range. We further demonstrated that several REX variants bind to human leukemic cell lines K-562, THP-1 and Jurkat, and this binding correlated with IL-23R cell-surface expression. The REX125, REX009 and REX128 variants competed with the p19 protein for binding to THP-1 cells. Moreover, the presence of REX125, REX009 and REX115 variants significantly inhibited the IL-23-driven expansion of IL-17-producing primary human CD4(+) T-cells. Thus, we conclude that unique IL-23R antagonists derived from the ABD scaffold were generated that might be useful in designing novel anti-inflammatory biologicals.


Assuntos
Anti-Inflamatórios/farmacologia , Fragmentos de Peptídeos/farmacologia , Receptores de Interleucina/antagonistas & inibidores , Células Th17/efeitos dos fármacos , Sequência de Aminoácidos , Anti-Inflamatórios/química , Proteínas de Bactérias/química , Ligação Competitiva , Avaliação Pré-Clínica de Medicamentos , Humanos , Fatores Imunológicos/química , Fatores Imunológicos/farmacologia , Interleucina-23/química , Interleucina-23/fisiologia , Células Jurkat , Células K562 , Dados de Sequência Molecular , Fragmentos de Peptídeos/química , Ligação Proteica , Receptores de Interleucina/fisiologia , Homologia de Sequência de Aminoácidos , Células Th17/metabolismo
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