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1.
Cytokine ; 78: 16-9, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26615568

RESUMO

Ocular surface inflammation is one of the primary mechanisms associated with dysfunctional tear syndrome (DTS), also known as dry eye disease. DTS, more prevalent in older populations, causes ocular discomfort and visual disturbance due to dryness on the surface layer in the eye. We used human conjunctival fibroblast cultures (HCJVF) to investigate the effects of inflammatory cytokines IFN-γ, TNF-α and IL-1ß (ITI) on the secretions of VEGF and chemokines. Our results demonstrate the elevated secretion of angiogenic VEGF molecules by ITI without affecting anti-angiogenic molecules, PEDF, endostatin, thrombospondin and sVEGF-R1. The secretion of interferon-γ inducible chemokines, CXCL9, -10, -11 by HCJVF were significantly enhanced by ITI. Our in vitro study supports previously reported observations of elevated VEGF and chemokines in tear fluids of DTS patients, reiterating the role of inflammatory reactions in DTS.


Assuntos
Quimiocinas/metabolismo , Citocinas/metabolismo , Síndromes do Olho Seco/metabolismo , Fibroblastos/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Idoso , Idoso de 80 Anos ou mais , Células Cultivadas , Quimiocinas/genética , Túnica Conjuntiva/citologia , Citocinas/genética , Síndromes do Olho Seco/imunologia , Regulação da Expressão Gênica , Humanos , Inflamação , Interferon gama/farmacologia , Interleucina-1beta/farmacologia , Lágrimas/química , Lágrimas/imunologia , Fator de Necrose Tumoral alfa/farmacologia
2.
Cancer Cytopathol ; 132(1): 50-59, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37812596

RESUMO

BACKGROUND: Cytokines are known to be a key a factor in numerous malignancies and to exert an important regulatory role in the tumor microenvironment. Interest has grown in understanding how cytokines modulate the tumor microenvironment and which cytokines may serve as markers of the tumor process; however, a complete picture of the cytokine landscape in bladder cancer remains unclear. METHODS: Fresh urine specimens with sufficient volume were collected at random intervals. The urine concentrations of IL-8 (CXCL8), CCL18, and CXCL9 were determined using the standard commercially available enzyme immunoassay. The urine concentrations of IL-6 were determined using the high sensitivity enzyme immunoassay kit. Urinary cytokine concentrations were normalized with urinary creatinine concentrations. RESULTS: Significantly elevated concentrations of IL-6 and IL-8 were detected in the urine from patients with urothelial carcinoma on follow-up compared to patients with benign follow-up. The presence of both IL-6 and IL-8 in the urine samples from the high grade urothelial carcinoma (HGUC) cohort revealed a clear discrimination when compared to samples from patients with benign follow-up. The presence of the combination of both IL-6 and IL-8 had a sensitivity of 90.0% and a specificity of 81.25%. Similar data were obtained when receiver operating characteristic analysis was performed on both IL-6 and IL-8 concentrations in the urine from patients with HGUC vs. the hematuria cohort. CONCLUSIONS: The presence of IL-6 and IL-8 in urine specimens may have predictive value for urothelial carcinoma. However, a large longitudinal study is required to statistically eliminate confounding factors and support this theory.


Assuntos
Carcinoma de Células de Transição , Neoplasias da Bexiga Urinária , Humanos , Carcinoma de Células de Transição/diagnóstico , Carcinoma de Células de Transição/patologia , Interleucina-6 , Interleucina-8 , Projetos Piloto , Microambiente Tumoral , Bexiga Urinária/patologia , Neoplasias da Bexiga Urinária/patologia , Urina , Urotélio/patologia
3.
Mol Vis ; 19: 737-50, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23592910

RESUMO

PURPOSE: The inflammatory response of the retinal pigment epithelium (RPE) is implicated in the pathogenesis of age-related macular degeneration. The microRNAs miR-146a and miR-146b-5p can regulate the inflammatory process by attenuating cytokine signaling via the nuclear factor-κB pathway. The aim of the present study is to investigate the expression of miR-146a and miR-146b-5p in human RPE cells and their response to proinflammatory cytokines. METHODS: Confluent cultures of RPE cells established from adult human donor eyes were treated with the proinflammatory cytokines interferon (IFN)-γ, tumor necrosis factor (TNF)-α, and interleukin (IL)-1ß. The expression of microRNAs was analyzed by real-time PCR using total RNA fraction. The retinal pigment epithelial cell line ARPE-19 was employed to analyze the promoter activity of the genes encoding miR-146a and miR-146b-5p. STAT1-binding activity of oligonucleotides was analyzed by electrophoretic mobility shift assay. ARPE-19 cells were transiently transfected with miR-146a and miR-146b-5p mimics for the analysis of IRAK1 expression by western immunoblotting. RESULTS: Real-time PCR analysis showed that miR-146a and 146b-5p are expressed in RPE cells. The cells responded to proinflammatory cytokines (IFN-γ + TNF-α + IL-1ß) by highly increasing the expression of both miR-146a and miR-146b-5p. This was associated with an increase in the expression of transcripts for CCL2, CCL5, CXCL9, CXCL10, and IL-6, and a decrease in that for HMOX1. The miR-146a induction was more dependent on IL-1ß, since its omission from the cytokine mix resulted in a greatly reduced response. Similarly, the induction of miR-146b-5p was more dependent on IFN-γ, since its omission from the cytokine mix minimized the effect. In addition, the increase in MIR146B promoter activity by the cytokine mix was effectively blocked by JAK inhibitor 1, a known inhibitor of the JAK/STAT signaling pathway. The expression of IRAK1 protein was decreased when ARPE-19 cells were transiently transfected with either miR-146a mimic or miR-146b-5p mimic. CONCLUSIONS: Our results clearly show that both miR-146a and miR-146b-5p are expressed in human RPE cells in culture and their expression is highly induced by proinflammatory cytokines (IFN-γ + TNF-α + IL-1ß). The induction of miR-146a showed a dependency on IL-1ß, while that of miR-146b-5p on IFN-γ. Our results show for the first time that miR-146b-5p expression is regulated by IFN-γ, potentially via the JAK/STAT pathway. These two microRNAs could play a role in inflammatory processes underlying age-related macular degeneration or other retinal degenerative diseases through their ability to negatively regulate the nuclear factor-κB pathway by targeting the expression of IRAK1.


Assuntos
Células Epiteliais/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Interferon gama/farmacologia , Interleucina-1beta/farmacologia , MicroRNAs/genética , Epitélio Pigmentado da Retina/citologia , Fator de Necrose Tumoral alfa/farmacologia , Adulto , Sequência de Bases , Ensaio de Desvio de Mobilidade Eletroforética , Células Epiteliais/efeitos dos fármacos , Perfilação da Expressão Gênica , Humanos , Mediadores da Inflamação/farmacologia , Quinases Associadas a Receptores de Interleucina-1/genética , Quinases Associadas a Receptores de Interleucina-1/metabolismo , MicroRNAs/metabolismo , Modelos Biológicos , Dados de Sequência Molecular , Regiões Promotoras Genéticas/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator de Transcrição STAT1/metabolismo , Fatores de Tempo
4.
Cytokine ; 61(3): 724-7, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23357298

RESUMO

In a microarray analysis of human retinal pigment epithelial cells (HRPE) treated with TGF-ß, in addition to the alteration of a number of known Extracellular matrix (ECM)-related genes regulated by TGF-ß, we found a significant increase in the expression of Kallmann Syndrome (KAL)-1 gene, that codes for the protein anosmin-1. Enhanced expression of KAL-1 by TGF-ß was validated by real-time PCR analysis. In in vitro experiments, TGF-ß receptor inhibitor abolished TGF-ß-induced expression of KAL-1. Immunofluorescence staining showed increased presence of anosmin-1 in TGF-ß treated HRPE cells, with distinct localization at the intercellular junctions. Treatment of HRPE cells with TGF-ß enhanced secretion of anosmin-1 and the release of anosmin-1 was further augmented by heparin sulfate. Enhanced secretion of anosmin-1 in the presence of TGF-ß and heparin was also observed in other ocular cells such as corneal epithelial and corneal fibroblast cultures. The role of anosmin-1, a protein with adhesion functions, in retinal structure, function and pathology has not been known and remains to be investigated.


Assuntos
Células Epiteliais/metabolismo , Proteínas da Matriz Extracelular/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Proteínas do Tecido Nervoso/genética , Epitélio Pigmentado Ocular/citologia , Fator de Crescimento Transformador beta/farmacologia , Células Epiteliais/efeitos dos fármacos , Proteínas da Matriz Extracelular/metabolismo , Imunofluorescência , Humanos , Immunoblotting , Proteínas do Tecido Nervoso/metabolismo , Reação em Cadeia da Polimerase em Tempo Real
5.
J Cell Physiol ; 227(1): 116-26, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-21374591

RESUMO

Chronic inflammation is implicated in the pathogenesis of age-related macular degeneration (AMD). Choroidal neovascularization (CNV) observed in exudative form of AMD results in vision loss. Human retinal pigment epithelial cell (HRPE) layer and choroidal tissue are the primary pathological sites in AMD. Pathological and therapeutic evidences have strongly indicated the vascular endothelial growth factor (VEGF) molecules as critical components in CNV pathogenesis. In these studies, we used human primary HRPE and choroidal fibroblast cells (HCHF) prepared from adult donor eyes. The effects of inflammatory cytokine (IFN-γ+ TNF-α+IL-1ß) mix (ICM) on global gene expression profiles in HRPE cells, revealed 10- and 9-fold increase in VEGF-A and VEGF-C expression, respectively. The microarray results were validated by quantitative RT-PCR and secretion of VEGFs proteins. IL-1ß is the most potent in inducing VEGFs secretion followed by IFN-γ and TNF-α, and the secretion was more effective in the presence of 2 and 3 cytokines. NF-κB and JAK-STAT pathway, but not HIF-1α, Sp-1, Sp-3, and STAT-3, transcription factors were upregulated and translocated to nucleus by ICM treatment. The mRNA levels of VEGF-A and VEGF-C and secretion of these proteins were also significantly enhanced by ICM in HCHF cells. The secretion of other angiogenic molecules, PEDF, SDF-1α, endostatin, and angiopoietins was not affected by ICM. Our results show that the inflammatory cytokines enhance secretion of VEGF-A and VEGF-C by HRPE and HCHF cells. These studies indicate that VEGFs secreted by these cells initiate and promote pathological choroidal and retinal noevascularization processes in AMD.


Assuntos
Citocinas/metabolismo , Inflamação/metabolismo , Degeneração Macular/metabolismo , Neovascularização Patológica/metabolismo , Epitélio Pigmentado da Retina/metabolismo , Fator A de Crescimento do Endotélio Vascular/biossíntese , Fator C de Crescimento do Endotélio Vascular/biossíntese , Células Cultivadas , Corioide/citologia , Corioide/metabolismo , Fibroblastos/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/fisiologia , Humanos , Inflamação/complicações , Inflamação/patologia , Degeneração Macular/etiologia , Degeneração Macular/patologia , Neovascularização Patológica/genética , Neovascularização Patológica/patologia , Análise de Sequência com Séries de Oligonucleotídeos , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fator A de Crescimento do Endotélio Vascular/genética , Fator C de Crescimento do Endotélio Vascular/genética
6.
Arch Virol ; 157(7): 1377-81, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22527863

RESUMO

The role of PMNs (neutrophils) in corneal herpes was studied using an in vitro system. Human corneal cells (HCE) and macrophages (THP-1) infected with HSV-1 or treated with virus components (DNA or virus immune complexes) released chemokines, which attracted PMNs. Highly reactive oxygen species were detected in PMNs. PMNs inhibited HSV when overlaid onto infected HCE cells (50:1). PMNs incubated with the supernatants of HCE cells treated with virus components released H(2)O(2) and myeloperoxidase. These inhibited virus growth. PMNs released NO and MIG, which may differentiate CD4 T cells to Th1. PMNs participate in innate immune responses, limit virus growth, and initiate immunopathology.


Assuntos
Meios de Cultivo Condicionados/farmacologia , Epitélio Corneano/citologia , Herpesvirus Humano 1/fisiologia , Leucócitos Mononucleares/metabolismo , Macrófagos/metabolismo , Neutrófilos/efeitos dos fármacos , Células Cultivadas , Quimiotaxia , Humanos , Peróxido de Hidrogênio/metabolismo , Neutrófilos/fisiologia , Peroxidase/metabolismo , Replicação Viral/efeitos dos fármacos
7.
Infect Immun ; 79(4): 1750-8, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21245265

RESUMO

Cerebral malaria (CM) is a major complication of Plasmodium falciparum infection, particularly in children. The pathogenesis of cerebral malaria involves parasitized red blood cell (RBC)-mediated vascular inflammation, immune stimulation, loss of blood-brain barrier integrity, and obstruction of cerebral capillaries. Therefore, blunting vascular inflammation and immune cell recruitment is crucial in limiting the disease course. Beta interferon (IFN-ß) has been used in the treatment of diseases, such as multiple sclerosis (MS) but has not yet been explored in the treatment of CM. Therefore, we sought to determine whether IFN-ß also limits disease progression in experimental cerebral malaria (ECM). Plasmodium berghei-infected mice treated with IFN-ß died later and showed increased survival, with improved blood-brain barrier function, compared to infected mice. IFN-ß did not alter systemic parasitemia. However, we identified multiple action sites that were modified by IFN-ß administration. P. berghei infection resulted in increased expression of chemokine (C-X-C motif) ligand 9 (CXCL9) in brain vascular endothelial cells that attract T cells to the brain, as well as increased T-cell chemokine (C-X-C motif) receptor 3 (CXCR3) expression. The infection also increased the cellular content of intercellular adhesion molecule 1 (ICAM-1), a molecule important for attachment of parasitized RBCs to the endothelial cell. In this article, we report that IFN-ß treatment leads to reduction of CXCL9 and ICAM-1 in the brain, reduction of T-cell CXCR3 expression, and downregulation of serum tumor necrosis factor alpha (TNF-α). In addition, IFN-ß-treated P. berghei-infected mice also had fewer brain T-cell infiltrates, further demonstrating its protective effects. Hence, IFN-ß has important anti-inflammatory properties that ameliorate the severity of ECM and prolong mouse survival.


Assuntos
Encéfalo/efeitos dos fármacos , Fatores Imunológicos/farmacologia , Interferon beta/uso terapêutico , Malária Cerebral/tratamento farmacológico , Animais , Barreira Hematoencefálica/efeitos dos fármacos , Barreira Hematoencefálica/metabolismo , Encéfalo/imunologia , Encéfalo/metabolismo , Quimiocina CXCL9/biossíntese , Quimiotaxia de Leucócito/efeitos dos fármacos , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Imuno-Histoquímica , Molécula 1 de Adesão Intercelular/biossíntese , Malária Cerebral/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Plasmodium berghei , Receptores CXCR3/biossíntese , Linfócitos T/efeitos dos fármacos , Linfócitos T/imunologia , Linfócitos T/metabolismo , Fator de Necrose Tumoral alfa/sangue
8.
Biochem Biophys Res Commun ; 391(1): 287-92, 2010 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-19913506

RESUMO

Interleukin-11 (IL-11) is an anti-apoptotic, anti-inflammatory cytokine with hematopoietic potential. The expression and protective actions of IL-11 have not been explored in the eye. The expression of IL-11 in primary cultures of human retinal pigment epithelial (HRPE) and human corneal fibroblast (HCRF) cells were evaluated in these studies. Constitutive secretion of IL-11 was not observed in either HRPE or HCRF. TNF-alpha+IL-1 induced IL-11 secretion and this production was inhibited by NFkappaB pathway inhibitors. IFN-gamma significantly inhibited TNF-alpha and IL-1 induced IL-11 secretion and inhibitors of JAK-STAT pathway reversed this inhibition. TGF-beta induced IL-11 secretion that was blocked by TGF-beta receptor 1 inhibitor but not by IFN-gamma. RT-PCR analysis confirmed the effects of IL-1, TNF-alpha, IFN-gamma and TGF-beta on IL-11 secretion at mRNA levels. Our results demonstrate that IL-11 is dramatically up regulated in retina and cornea cells and that IFN-gamma is a physiological inhibitor of IL-11 expression.


Assuntos
Córnea/imunologia , Regulação da Expressão Gênica , Interferon gama/metabolismo , Interleucina-11/genética , Retina/imunologia , Linhagem Celular , Córnea/citologia , Humanos , Interferon gama/farmacologia , Interleucina-1/metabolismo , Interleucina-1/farmacologia , Interleucina-11/antagonistas & inibidores , Interleucina-11/metabolismo , Janus Quinases/antagonistas & inibidores , Janus Quinases/metabolismo , NF-kappa B/antagonistas & inibidores , NF-kappa B/metabolismo , Retina/citologia , Fatores de Transcrição STAT/antagonistas & inibidores , Fatores de Transcrição STAT/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Fator de Necrose Tumoral alfa/farmacologia
9.
Biochem Biophys Res Commun ; 402(2): 390-5, 2010 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-20950585

RESUMO

Inflammatory response of the retinal pigment epithelium plays a critical role in the pathogenesis of retinal degenerative diseases such as age-related macular degeneration. Our previous studies have shown that human retinal pigment epithelial (HRPE) cells, established from adult donor eyes, respond to inflammatory cytokines by enhancing the expression of a number of cytokines and chemokines. To investigate the role of microRNA (miRNA) in regulating this response, we performed microarray analysis of miRNA expression in HRPE cells exposed to inflammatory cytokine mix (IFN-γ+TNF-α+IL-1ß). Microarray analysis revealed ∼11-fold increase in miR-155 expression, which was validated by real-time PCR analysis. The miR-155 expression was enhanced when the cells were treated individually with IFN-γ, TNF-α or IL-1ß, but combinations of the cytokines exaggerated the effect. The increase in miR-155 expression by the inflammatory cytokines was associated with an increase in STAT1 activation as well as an increase in protein binding to putative STAT1 binding elements present in the MIR155 gene promoter region. All these activities were effectively blocked by JAK inhibitor 1. Our results show that the inflammatory cytokines increase miR-155 expression in human retinal pigment epithelial cells by activating the JAK/STAT signaling pathway.


Assuntos
Citocinas/metabolismo , Janus Quinases/biossíntese , MicroRNAs/biossíntese , Epitélio Pigmentado da Retina/metabolismo , Fator de Transcrição STAT1/metabolismo , Células Cultivadas , Humanos , Regiões Promotoras Genéticas
10.
Pharm Res ; 26(5): 1226-35, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-18781380

RESUMO

PURPOSE: To investigate whether conjunctival epithelial cells express transport processes for opioid peptides. METHODS: We monitored the uptake of [(3)H]deltorphin II and [(3)H]DADLE, two hydrolysis-resistant synthetic opioid peptides, in the rabbit conjunctival epithelial cell line CJVE and elucidated the characteristics of the uptake process. RESULTS: CJVE cells express robust uptake activity for deltorphin II and DADLE. Both opioid peptides compete with each other for transport. Several endogenous and synthetic opioid peptides, but not non-peptide opioid antagonists, are recognized by the transport process. Though various peptides inhibit the uptake of deltorphin II and DADLE in a similar manner, the uptake of deltorphin II is partly Na(+)-dependent whereas that of DADLE mostly Na(+)-independent. The transport process shows high affinity for many endogenous/synthetic opioid peptides. Functional features reveal that this transport process may be distinct from the opioid peptide transport system described in the retinal pigment epithelial cell line ARPE-19 and also from the organic anion transporting polypeptides, which are known to transport opioid peptides. CONCLUSIONS: CJVE cells express a novel, hitherto unknown transport process for endogenous/synthetic opioid peptides. This new transport process may offer an effective delivery route for opioid peptide drugs to the posterior segment of the eye.


Assuntos
Transporte Biológico/efeitos dos fármacos , Túnica Conjuntiva/citologia , Células Epiteliais/metabolismo , Peptídeos Opioides/farmacocinética , Amilorida/análogos & derivados , Amilorida/farmacologia , Animais , Linhagem Celular , Leucina Encefalina-2-Alanina/farmacocinética , Naloxona/farmacologia , Oligopeptídeos/farmacocinética , Transportadores de Ânions Orgânicos/antagonistas & inibidores , Coelhos , ATPase Trocadora de Sódio-Potássio/antagonistas & inibidores
11.
Arch Virol ; 154(2): 219-26, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19115032

RESUMO

Angiogenesis and inflammatory mediators are critical pathogenic factors in herpetic stromal keratitis (HSK). Since disease progresses without infectious virus, HSV-DNA and HSV-IgG complexes (HSV-IC) may contribute to HSK by triggering these factors. Production of VEGF and MMP-9 was studied in vitro using corneal epithelial cells (HCE), fibroblasts (HCRF) and macrophages (THP-1). VEGF was elevated in HCRF and THP-1 following treatment with HSV-DNA and HSV-IC. MMP-9 was elevated in THP-1 but not in corneal cells. When anti-HSV-IgG(Fab')2 complexes stimulated THP-1, MMP-9 was reduced to control levels. Pretreatment of THP-1 with anti-TLR-2 and -3 inhibited MMP-9 production. Thus, HSV-IC may stimulate THP-1 through the Fc receptor and TLRs. Proinflammatory cytokines (IL-1b, IL-6, and TNF-alpha) increased VEGF and MMP-9 in corneal cells and macrophages. These studies indicate that the continued presence of HSV-DNA and HSV-IC contribute to angiogenesis and inflammation in HSK. Thus, cytokines and TLRs may be potential targets for intervention.


Assuntos
Complexo Antígeno-Anticorpo/imunologia , Neovascularização da Córnea/imunologia , Herpesvirus Humano 1/imunologia , Ceratite Herpética/imunologia , Metaloproteinase 9 da Matriz/biossíntese , Fator A de Crescimento do Endotélio Vascular/biossíntese , Anticorpos Antivirais/imunologia , Complexo Antígeno-Anticorpo/farmacologia , Células Cultivadas , Córnea/imunologia , Córnea/virologia , Neovascularização da Córnea/virologia , Citocinas/farmacologia , DNA Viral/imunologia , DNA Viral/farmacologia , Fibroblastos/metabolismo , Fibroblastos/virologia , Herpesvirus Humano 1/genética , Humanos , Imunoglobulina G/imunologia , Ceratite Herpética/complicações , Ceratite Herpética/virologia , Macrófagos/imunologia , Macrófagos/virologia , Testes de Neutralização
12.
Biochem Biophys Res Commun ; 374(3): 479-84, 2008 Sep 26.
Artigo em Inglês | MEDLINE | ID: mdl-18639520

RESUMO

Inflammatory processes within the cornea are known to be associated with corneal neovascularization (CN). We examined the effects of inflammatory mediators on the expression of angiogenic factors by corneal cells. TNF-alpha and IL-1 induced VEGF-A secretion by corneal fibroblasts (HCRF) and this was inhibited significantly by IFN-gamma. Constitutively secreted VEGF-A by corneal epithelial cells (HCE) was not affected by these cytokines. Moreover, sVEGF-R1(sFlt-1) secretion by HCRF was stimulated significantly by IFN-gamma. JAK-STAT pathway inhibitor reversed the effects of IFN-gamma on VEGF-A and sFlt-1 secretion by HCRF. RT-PCR analysis showed that IFN-gamma influences the expression of VEGF-A and sFlt-1 by affecting their mRNA level. IFN-gamma inhibited TGF-beta induced VEGF-A secretion but not sVEGF-R1 secretion. This is the first report demonstrating the inhibitory and stimulatory effects of IFN-gamma on VEGF-A and sFlt-1 secretion, respectively. Our results suggest that IFN-gamma acts as an anti-angiogenic cytokine in the human cornea.


Assuntos
Córnea/enzimologia , Neovascularização da Córnea/enzimologia , Interferon gama/fisiologia , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/agonistas , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Linhagem Celular , Córnea/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Fibroblastos/enzimologia , Humanos , Interferon gama/farmacologia , Interleucina-1/farmacologia , Janus Quinase 1/antagonistas & inibidores , RNA Mensageiro/biossíntese , Receptor Tipo 1 de Fator de Crescimento de Fibroblastos/metabolismo , Fator de Crescimento Transformador beta/farmacologia , Fator A de Crescimento do Endotélio Vascular/metabolismo
13.
J Neuroimmunol ; 193(1-2): 28-37, 2008 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-18037505

RESUMO

Mouse hepatitis virus induces a biphasic disease in BALB/c mice that consists of an acute retinitis followed by progression to a chronic retinal degeneration with autoimmune reactivity. Retinal degeneration resistant CD-1 mice do not develop the late phase. What host factors contribute to the distinct responses to the virus are unknown. Herein, we show that IFN-alpha, IFN-beta and IFN-gamma act in concert as part of the innate immune response to the retinal infection. At day 2, high serum levels of IFN-gamma, CXCL9 and CXCL10, were detected in BALB/c mice. Moreover, elevated levels of CXCL9 and CXCL10 gene expression were detected in retinal tissue. Although IFN-gamma and the chemokines were detected in CD-1 mice, they were at significantly lower levels compared to BALB/c mice. These augmented innate responses observed correlated with the development of autoimmune reactivity and retinal degeneration and thus may contribute to the pathogenic processes.


Assuntos
Quimiocina CXCL10/biossíntese , Quimiocina CXCL9/biossíntese , Infecções por Coronavirus/imunologia , Interferons/biossíntese , Vírus da Hepatite Murina , Degeneração Retiniana/etiologia , Animais , Quimiocina CXCL10/sangue , Quimiocina CXCL10/genética , Quimiocina CXCL9/sangue , Quimiocina CXCL9/genética , Infecções por Coronavirus/complicações , Imunidade Inata , Interferons/sangue , Interferons/genética , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Degeneração Retiniana/imunologia , Síndrome Respiratória Aguda Grave/imunologia
14.
Semin Ophthalmol ; 23(4): 229-34, 2008.
Artigo em Inglês | MEDLINE | ID: mdl-18584560

RESUMO

In the herpetic stromal keratitis (HSK), HSV DNA fragments and HSV-IgG immune complexes (HSV-IC) are present in most of the corneas long after infective virus has disappeared. These viral components are highly immunogenic and potentiate production of proinflammatory cytokines and chemokines via Toll-like receptors (TLRs) expressed on the corneal cells and macrophages. In addition angiogenic factors, such as vascular endothelium growth factor (VEGF) and the tissue damaging enzyme matrix metalloproteinase 9 (MMP-9) deeply involved in the pathogenesis of HSK, are also induced by corneal cells and macrophages through the recognition of these viral components. These processes elicited by residual viral DNA and HSV-IC are likely one of the sustained driving force in the development of HSK. Hence, strategies developed to alter these pathways should lead to new preventative and therapeutic measures.


Assuntos
Substância Própria/virologia , Ceratite Herpética/etiologia , Simplexvirus/genética , Simplexvirus/imunologia , Animais , DNA Viral/genética , Humanos , Imunoglobulina G/imunologia , Receptores Toll-Like/fisiologia
15.
J Neuroimmunol ; 316: 74-79, 2018 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-29310941

RESUMO

Autoimmune retinopathy (AIR) is a rare immune-mediated retinopathy associated with circulating antiretinal antibodies (ARAs). Other prominent features of AIR include visual field deficits and photoreceptor dysfunction in the setting of progressive unexplained vision loss. The role of inflammation is poorly understood in AIR. Since cytokines play a central role in the initiation and development of inflammation, we evaluated the presence of proinflammatory cytokines and chemokines in AIR patient sera. We demonstrate that IL-6 and CXCL9 are both elevated in AIR patient sera. Moreover, the presence and concentration of these 2 molecules appear to correlate with AIR patient disease severity. This cytokine profile, IL-6 and CXCL9, has been described to participate in a variety of autoimmune and inflammatory diseases. Our study provides support for an activated inflammatory process in AIR and identifies possible mechanisms that can drive autoimmunity in this disease.


Assuntos
Doenças Autoimunes/imunologia , Quimiocina CXCL9/sangue , Interleucina-6/sangue , Doenças Retinianas/imunologia , Adulto , Idoso , Doenças Autoimunes/sangue , Quimiocina CXCL9/imunologia , Feminino , Humanos , Interleucina-6/imunologia , Masculino , Pessoa de Meia-Idade , Doenças Retinianas/sangue
16.
Microbes Infect ; 8(8): 2236-44, 2006 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-16782382

RESUMO

Cytomegalovirus (CMV) retinitis is characterized by alterations in retinal cell function and host responses to virus replication. The goal of this study was to evaluate the induction of cyclooxygenase-2 (COX-2) and prostaglandin (PGE) in CMV infected human retinal pigment epithelial (RPE) cells and to determine their effect on virus replication. CMV immediate early (IE) protein and COX-2 proteins were identified in RPE cells in retinal tissue sections from patients with CMV retinitis. COX-2 mRNA and protein were induced after CMV infection of human RPE cell cultures. CMV infection of RPE cells induced translocation of NF-kappaB from the cytoplasm to the nucleus. PGE1 and PGE2 were significantly (p<0.001) increased in human RPE cell cultures infected with CMV. Inhibition of CMV IE gene by antisense oligonucleotides abrogated induction of mRNA for COX-2 and protein synthesis of COX-2 and PGE2. PGE enhanced CMV plaque formation and real time PCR analysis revealed that PGE treatment significantly increased CMV DNA copy numbers. These studies demonstrate that when CMV replicates within human RPE cells, COX-2 induction augments virus replication via the PGE pathway. The induction of COX-2 and PGE during retinal CMV infection may augment virus replication and alter a variety of retinal physiological responses.


Assuntos
Ciclo-Oxigenase 2/biossíntese , Citomegalovirus/fisiologia , Epitélio Pigmentado Ocular/virologia , Prostaglandinas/biossíntese , Replicação Viral , Núcleo Celular/química , Células Cultivadas , Ciclo-Oxigenase 2/genética , Citomegalovirus/crescimento & desenvolvimento , Retinite por Citomegalovirus/patologia , Retinite por Citomegalovirus/virologia , Citoplasma/química , DNA Viral/análise , Expressão Gênica , Humanos , Proteínas Imediatamente Precoces/análise , Proteínas Imediatamente Precoces/antagonistas & inibidores , Imuno-Histoquímica , Microscopia de Fluorescência , NF-kappa B/metabolismo , Oligonucleotídeos Antissenso/genética , RNA Mensageiro/análise , RNA Mensageiro/genética , Retina/química , Ensaio de Placa Viral , Proteínas Virais/análise
17.
Cornea ; 25(6): 709-14, 2006 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-17077666

RESUMO

PURPOSE: To evaluate the modulatory effects of anti-inflammatory agents, dexamethasone (Dex) and cyclosporin A (CsA), on the production of cytokines and chemokines by human corneal cells in vitro following stimulation by the pro-inflammatory cytokine after interleukin 1beta (IL-1beta). METHODS: A human corneal epithelial (HCE) cell line and human corneal fibroblasts (HCFs) were stimulated in culture with IL-1beta and treated with Dex or CsA. The gene expression for selected cytokines and chemokines was determined by reverse transcriptase-polymerase chain reaction (RT-PCR). The secretion of cytokines and chemokines was measured by enzyme-linked immunosorbent assay. RESULTS: IL-1beta enhanced the mRNA and/or protein levels of granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-8, and monocyte chemotactic protein (MCP)-1 in HCE and IL-6, IL-8, MCP-3, and regulated on T-cell activation expressed secreted (RANTES) in HCFs. Treatment with CsA did not inhibit cytokine production in either HCE or HCFs. In contrast, Dex treatment inhibited the IL-1beta-induced production of GM-CSF, IL-6, IL-8, MCP-3, and RANTES, but not MCP-1. CONCLUSION: These results show that Dex, but not CsA, has direct immunosuppressive effects on the resident corneal cells, HCE and HCFs. This suggests that the clinically observed immunosuppressive effects of topical CsA are mediated primarily through the immune cells.


Assuntos
Anti-Inflamatórios/farmacologia , Ciclosporina/farmacologia , Citocinas/genética , Dexametasona/farmacologia , Epitélio Corneano/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Imunossupressores/farmacologia , Técnicas de Cultura de Células , Quimiocina CCL2/genética , Quimiocina CCL5/genética , Quimiocina CCL7 , Substância Própria/citologia , Epitélio Corneano/metabolismo , Fibroblastos/metabolismo , Expressão Gênica/efeitos dos fármacos , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Humanos , Interleucina-1beta/farmacologia , Interleucina-6/genética , Interleucina-8/genética , Proteínas Quimioatraentes de Monócitos/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa
18.
Am J Ophthalmol ; 168: 183-190, 2016 08.
Artigo em Inglês | MEDLINE | ID: mdl-27210277

RESUMO

PURPOSE: To develop diagnostic criteria for nonparaneoplastic autoimmune retinopathy (AIR) through expert panel consensus and to examine treatment patterns among clinical experts. DESIGN: Modified Delphi process. METHODS: A survey of uveitis specialists in the American Uveitis Society, a face-to-face meeting (AIR Workshop) held at the National Eye Institute, and 2 iterations of expert panel surveys were used in a modified Delphi process. The expert panel consisted of 17 experts, including uveitis specialists and researchers with expertise in antiretinal antibody detection. Supermajority consensus was used and defined as 75% of experts in agreement. RESULTS: There was unanimous agreement among experts regarding the categorization of autoimmune retinopathies as nonparaneoplastic and paraneoplastic, including cancer-associated retinopathy and melanoma-associated retinopathy. Diagnostic criteria and tests essential to the diagnosis of nonparaneoplastic AIR and multiple supportive criteria reached consensus. For treatment, experts agreed that corticosteroids and conventional immunosuppressives should be used (prescribed) as first- or second-line treatments, though a consensus agreed that biologics and intravenous immunoglobulin were considered appropriate in the treatment of nonparaneoplastic AIR patients regardless of the stage of disease. Experts agreed that more evidence is needed to treat nonparaneoplastic AIR patients with long-term immunomodulatory therapy and that there is enough equipoise to justify randomized, placebo-controlled trials to determine if nonparaneoplastic AIR patients should be treated with long-term immunomodulatory therapy. Regarding antiretinal antibody detection, consensus agreed that a standardized assay system is needed to detect serum antiretinal antibodies. Consensus agreed that an ideal assay should have a 2-tier design and that Western blot and immunohistochemistry should be the methods used to identify antiretinal antibodies. CONCLUSIONS: Consensus was achieved using a modified Delphi process to develop diagnostic criteria for nonparaneoplastic AIR. There is enough equipoise to justify randomized, placebo-controlled trials to determine whether patients with nonparaneoplastic AIR should be treated with long-term immunomodulatory therapy. Efforts to develop a standardized 2-tier assay system for the detection of antiretinal antibodies have been initiated as a result of this study.


Assuntos
Doenças Autoimunes/diagnóstico , Doenças Retinianas/diagnóstico , Autoanticorpos/sangue , Autoantígenos/imunologia , Doenças Autoimunes/imunologia , Consenso , Técnica Delphi , Humanos , Síndromes Paraneoplásicas Oculares/diagnóstico , Retina/imunologia , Doenças Retinianas/imunologia
19.
J Neuroimmunol ; 166(1-2): 65-74, 2005 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-16039725

RESUMO

Experimental coronavirus retinopathy (ECOR) is a virally triggered model of retinal degeneration composed of both genetic and autoimmune components. Since TNF-alpha plays a role in immune-mediated processes we evaluated the levels of TNF-alpha/TNF-alpha receptors and the downstream signaling molecule nitric oxide (NO) during disease in both retinal degeneration susceptible BALB/c and degeneration resistant CD-1 mice. Following coronavirus injection, TNF-alpha mRNA was detected at higher levels within the retinas, and concentrations of TNF-alpha (p<0.005) and sTNFR1 (p<0.0005) proteins were increased within the sera of BALB/c but not CD-1 mice. While concentrations of sTNFR2 proteins were elevated in both BALB/c (p<0.00005) and CD-1 (p<0.005) mice compared to controls, concentrations were higher in BALB/c mice (p<0.0005). Gene expression of iNOS while initially high in BALB/c mice decreased during the acute phase of infection, while it increased in CD-1 mice. These trends are attributable to differences in monocyte TNFR2 release (p<0.0005) between the strains since sTNFR2 decreased (p<0.01) levels of NO production. These studies demonstrate that retinal degeneration following viral infection is associated with increased release of TNF-alpha/TNF receptors combined with a down-regulation of NO. Furthermore they suggest that these molecules are involved in alterations in immune response leading to autoimmune reactivity.


Assuntos
Infecções por Coronavirus/complicações , Receptores Tipo II do Fator de Necrose Tumoral/metabolismo , Degeneração Retiniana/virologia , Transdução de Sinais , Fator de Necrose Tumoral alfa/metabolismo , Animais , Infecções por Coronavirus/metabolismo , Expressão Gênica , Imuno-Histoquímica/métodos , Interferon-alfa/farmacologia , Interferon gama/farmacologia , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos , Monócitos/metabolismo , Vírus da Hepatite Murina , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase/genética , Óxido Nítrico Sintase Tipo II , Concentração Osmolar , Receptores Tipo I de Fatores de Necrose Tumoral/metabolismo , Receptores Tipo II do Fator de Necrose Tumoral/química , Retina/metabolismo , Solubilidade , Coloração e Rotulagem , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/farmacologia
20.
Aging Dis ; 6(6): 444-55, 2015 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-26618046

RESUMO

Chemokine reeptor-3 (CCR-3) was shown to be associated with choroidal neovascularization (CNV) in age-related macular degeneration (AMD). AMD is a vision threatening retinal disease that affects the aging population world-wide. Retinal pigment epithelium and choroid in the posterior part of the retina are the key tissues targeted in the pathogenesis of CNV in AMD. We used human retinal pigment epithelial (HRPE) and choroidal fibroblast (HCHF) cells, prepared from aged adult human donor eyes, to evaluate the expression of major CCR-3 ligands, CCL-5, CCL -7, CCL-11,CCL-24 and CCL-26. Microarray analysis of gene expression in HRPE cells treated with inflammatory cytokine mix (ICM= IFN-γ+TNF-α+IL-1ß) revealed 75 and 23-fold increase in CCL-5 and CCL-7 respectively, but not CCL-11, CCL-24 and CCL-26. Chemokine secretion studies of the production of CCL5 and CCL7 by HRPE corroborated with the gene expression analysis data. When the HRPE cells were treated with either individual cytokines or the ICM, both CCL-5 and CCL-7 were produced in a dose dependent manner. Similar to the gene expression data, the ICM did not enhance HRPE production of CCL-11, CCL-24 and CCL-26. CCL-11 and CCL-26 were increased with IL-4 treatment and this HRPE production was augmented in the presence of TNF-α and IL1ß. When HCHF cells were treated with either individual cytokines or the ICM, both CCL-5 and CCL-7 were produced in a dose dependent fashion. IL-4 induced low levels of CCL-11 and CCL-26 in HCHF and this production was significantly enhanced by TNF-α. Under these conditions, neither HRPE nor HCHF were demonstrated to produce CCL-24. These data demonstrate that chronic inflammation triggers CCL-5 and CCL-7 release by HRPE and HCHF and the subsequent interactions with CCR3 may participate in pathologic processes in AMD.

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