RESUMO
Purpose: Age-related macular degeneration (AMD) is the leading cause of blindness in Western populations. While an overactive complement system has been linked to pathogenesis, mechanisms contributing to its activation are largely unknown. In aged and AMD eyes, loss of the elastin layer (EL) of Bruch's membrane (BrM) has been reported. Elastin antibodies are elevated in patients with AMD, the pathogenic significance of which is unclear. Here we assess the role of elastin antibodies using a mouse model of smoke-induced ocular pathology (SIOP), which similarly demonstrates EL loss. Methods: C57BL/6J mice were immunized with elastin or elastin peptide oxidatively modified by cigarette smoke (ox-elastin). Mice were then exposed to cigarette smoke or air for 6 months. Visual function was assessed by optokinetic response, retinal morphology by spectral-domain optical coherence tomography and electron microscopy, and complement activation and antibody deposition by Western blot. Results: Ox-elastin IgG and IgM antibodies were elevated in ox-elastin immunized mice following 6 months of smoke, whereas elastin immunization had a smaller effect. Ox-elastin immunization exacerbated smoke-induced vision loss, with thicker BrM and more damaged retinal pigment epithelium (RPE) mitochondria compared with mice immunized with elastin or nonimmunized controls. These changes were correlated with increased levels of IgM, IgG2, IgG3, and complement activation products in RPE/choroid. Conclusions: These data demonstrate that SIOP mice generate elastin-specific antibodies and that immunization with ox-elastin exacerbates ocular pathology. Elastin antibodies represented complement fixing isotypes that, together with the increased presence of complement activation seen in immunized mice, suggest that elastin antibodies exert pathogenic effects through mediating complement activation.
Assuntos
Autoanticorpos/sangue , Lâmina Basilar da Corioide/patologia , Modelos Animais de Doenças , Elastina/imunologia , Atrofia Geográfica/etiologia , Epitélio Pigmentado da Retina/patologia , Fumar/efeitos adversos , Animais , Western Blotting , Ativação do Complemento/fisiologia , Proteínas do Sistema Complemento/metabolismo , Sensibilidades de Contraste/fisiologia , Ensaio de Imunoadsorção Enzimática , Atrofia Geográfica/imunologia , Atrofia Geográfica/patologia , Imunização , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Camundongos , Camundongos Endogâmicos C57BL , Nistagmo Optocinético/fisiologia , Oxirredução , Produtos do Tabaco , Acuidade Visual/fisiologiaRESUMO
PURPOSE: To evaluate the presence of interleukin-17 (IL-17)-producing cells in patients with geographic atrophy (GA). METHODS: In this short report, we analyzed IL-17, CD3, and IBA-1 expression by immunohistochemistry on paraffin-embedded sections from 13 donors with a known history of GA, confirmed by fundus appearance and histology, and 7 age-matched control donors. RESULTS/CONCLUSION: We showed that IL-17+ cells are found near areas of retinal pigmented epithelium atrophy in the eyes of GA patients. IL-17+ cells mainly localized to CD3+ cells, which identifies T lymphocytes, as well as IBA-1+ cells, which identifies mononuclear phagocytes. Therefore, IL-17 could be involved in the pathological mechanisms that contribute to the degeneration observed in GA.
Assuntos
Corioide/patologia , Atrofia Geográfica/metabolismo , Imunidade Celular , Interleucina-17/biossíntese , Macrófagos/metabolismo , Linfócitos T/metabolismo , Idoso de 80 Anos ou mais , Corioide/imunologia , Feminino , Angiofluoresceinografia , Fundo de Olho , Atrofia Geográfica/imunologia , Atrofia Geográfica/patologia , Humanos , Imuno-Histoquímica , Macrófagos/imunologia , Macrófagos/patologia , Masculino , Linfócitos T/imunologia , Linfócitos T/patologiaRESUMO
It is generally accepted that geographic atrophy (GA), a currently untreatable advanced form of age-related macular degeneration (AMD), is a multifactorial disease resulting in gradual and permanent blindness. Various risk factors are demonstrated to be responsible for its pathogenesis, such as aging, light exposure, and smoking. Molecular components associated with those risk factors form a complex and interwoven network at the confluence of inflammation, highlighting the significance of inflammasome activation in GA progression. Recently, a new type of modification in AMD microenvironment has been discovered, other than extensively-studied complement dysregulation, lipofuscin deposit, and oxidative by-products, to activate inflammasome. The accumulation of Alu RNA, resulting from DICER1 deficiency, is shown capable of triggering the activation of NLRP3 inflammasome and causing caspase-8-activated apoptosis in an IL-18/MyD88-dependent manner, which provides a new source of evidence for the interplay between cell death and inflammasome. In this review, we lay the emphasis on the mechanism by which Alu RNA activates NLRP3 inflammasome and downstream apoptotic proteins, and on its clinical relevance to GA and potential therapeutic approaches. We also point out several possible crosstalks among inflammasome and different acts of cell death which remain to be further investigated in Alu RNA-induced RPE degeneration.
Assuntos
Elementos Alu/imunologia , RNA Helicases DEAD-box/genética , Atrofia Geográfica/imunologia , Inflamassomos/imunologia , Terapia de Alvo Molecular/métodos , Proteína 3 que Contém Domínio de Pirina da Família NLR/imunologia , RNA não Traduzido/imunologia , Ribonuclease III/genética , Apoptose/imunologia , Proteínas Reguladoras de Apoptose/metabolismo , Atrofia Geográfica/genética , Atrofia Geográfica/terapia , Humanos , Lipofuscina/metabolismo , Sistema de Sinalização das MAP Quinases , Espécies Reativas de Oxigênio/imunologiaRESUMO
BACKGROUND: Age-related macular degeneration (AMD) is the leading cause of central vision loss in older adults. Anti-retinal autoantibodies (AAbs) have been found in individuals with AMD. The goal of the study was to determine the AAb specificity in different stages of AMD, and determine whether there is a prevalent AAb signature. METHODS: Sera of 134 participants in the Age-related Eye Disease Study were analyzed for anti-retinal AAbs by western blotting. The subjects were classified by diagnostic subgroups based upon their clinical classification: No AMD, Intermediate AMD, and Late AMD - geographic atrophy (GA) and Late AMD - neovascular (NV). RESULTS: The presence of anti-retinal AAb was detected in 58% patients with Intermediate and Late AMD, and 54% of those with no AMD. AAbs bound to fifteen different retinal antigens. Most individuals had 1 specific AAbs (67%), with the remainder having 2 to 4 different AAbs. Over 40% of patients with Intermediate AMD, and 46% of those with GA had anti-enolase AAbs, compared with 29% of individuals with NV and 29% with no AMD. Different AAbs signatures related to NV as compared to GA and/or Intermediate AMD were distinguished. Anti-40-kDa (10%) and 42-kDa (16%) autoantibodies were associated with Intermediate AMD, while anti-30-kDa AAbs (23%) were primarily present in GA. Anti-32-kDa (12%), 35-kDa (21%), and 60-kDa (8%) AAbs were more frequent in NV AMD. CONCLUSIONS: A unique AAb pattern for each of the disease subgroups was present when AMD progressed from the intermediate to the late forms of severity. Differences in the frequency of specific AAbs between AMD subgroups suggested that they may participate in pathogenicity of AMD. Further studies are necessary to confirm these observations in the larger cohort and individual AMD patients over time.
Assuntos
Autoanticorpos/sangue , Autoantígenos/imunologia , Atrofia Geográfica/imunologia , Retina/imunologia , Degeneração Macular Exsudativa/classificação , Degeneração Macular Exsudativa/imunologia , Idoso , Artrite/imunologia , Biomarcadores/sangue , Western Blotting , Feminino , Atrofia Geográfica/classificação , Humanos , Masculino , Prevalência , Fumar/imunologiaRESUMO
We have characterized a naturally-occurring mutation in mice that causes slow, progressive photoreceptor degeneration, white fundus flecks, and late-onset RPE atrophy. These animals predictably lose visual function as photoreceptors degenerate. Genetic studies identified a deletion in the 5' coding sequence of Mfrp, designated Mfrp (174delG) , which essentially results in a complete knockout at the protein level. We have shown in Mfrp (174delG) mice that these white fundus flecks are due to the presence of F4/80+ inflammatory cells in the subretinal space. Here we expand on our initial description of the cells with additional markers and by determining their origin. We have also begun an analysis of complement factors in the RPE and found decreased levels of C3d, suggesting that the alternative complement pathway may be misregulated. Finally, we compare and contrast the characteristics of fundus images in Mfrp (174delG) mice with those of other mutations that cause similar irregularities, including Crb1 (rd8) and RDH5, and discuss the structural differences that may underlie them.
Assuntos
Proteínas do Sistema Complemento/imunologia , Atrofia Geográfica/patologia , Degeneração Macular/patologia , Monócitos/patologia , Oxirredutases do Álcool/genética , Animais , Transplante de Medula Óssea , Via Alternativa do Complemento/imunologia , Modelos Animais de Doenças , Proteínas do Olho/genética , Atrofia Geográfica/genética , Atrofia Geográfica/imunologia , Degeneração Macular/genética , Degeneração Macular/imunologia , Proteínas de Membrana/genética , Camundongos , Camundongos Knockout , Monócitos/imunologia , Proteínas do Tecido Nervoso/genética , Retina/imunologia , Retina/patologiaRESUMO
AIMS: With ageing extracellular material is deposited in Bruch's membrane, as drusen. Lipofuscin is deposited in retinal pigment epithelial cells. Both of these changes are associated with age related macular degeneration, a disease now believed to involve chronic inflammation at the retinal-choroidal interface. We hypothesise that these molecules may act as danger signals, causing the production of inflammatory chemokines and cytokines by the retinal pigment epithelium, via activation of pattern recognition receptors. METHODS: ARPE-19 cells were stimulated in vitro with the following reported components of drusen: amyloid-ß (1-42), Carboxyethylpyrrole (CEP) modified proteins (CEP-HSA), Nε-(Carboxymethyl)lysine (CML) modified proteins and aggregated vitronectin. The cells were also stimulated with the major fluorophore of lipofuscin: N-retinylidene-N-retinylethanolamine (A2E). Inflammatory chemokine and cytokine production was assessed using Multiplex assays and ELISA. The mechanistic evaluation of the NLRP3 inflammasome pathway was assessed in a stepwise fashion. RESULTS: Of all the molecules tested only A2E induced inflammatory chemokine and cytokine production. 25 µM A2E induced the production of significantly increased levels of the chemokines IL-8, MCP-1, MCG and MIP-1α, the cytokines IL-1ß, IL-2, IL-6, and TNF-α, and the protein VEGF-A. The release of IL-1ß was studied further, and was determined to be due to NLRP3 inflammasome activation. The pathway of activation involved endocytosis of A2E, and the three inflammasome components NLRP3, ASC and activated caspase-1. Immunohistochemical staining of ABCA4 knockout mice, which show progressive accumulation of A2E levels with age, showed increased amounts of IL-1ß proximal to the retinal pigment epithelium. CONCLUSIONS: A2E has the ability to stimulate inflammatory chemokine and cytokine production by RPE cells. The pattern recognition receptor NLRP3 is involved in this process. This provides further evidence for the link between A2E, inflammation, and the pathogenesis of AMD. It also supports the recent discovery of NLRP3 inflammasome activation in AMD.
Assuntos
Proteínas de Transporte/metabolismo , Células Epiteliais/metabolismo , Inflamassomos/metabolismo , Interleucina-1beta/biossíntese , Retinoides/fisiologia , Transportadores de Cassetes de Ligação de ATP/deficiência , Transportadores de Cassetes de Ligação de ATP/genética , Animais , Proteínas de Transporte/genética , Linhagem Celular , Citoplasma/metabolismo , Endocitose , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/imunologia , Técnicas de Silenciamento de Genes , Atrofia Geográfica/imunologia , Atrofia Geográfica/patologia , Humanos , Mediadores da Inflamação/metabolismo , Interleucina-1beta/genética , Camundongos , Camundongos da Linhagem 129 , Camundongos Knockout , Proteína 3 que Contém Domínio de Pirina da Família NLR , Precursores de Proteínas/biossíntese , Precursores de Proteínas/genética , Transporte Proteico , RNA Interferente Pequeno/genética , Epitélio Pigmentado da Retina/imunologia , Epitélio Pigmentado da Retina/metabolismo , Epitélio Pigmentado da Retina/patologia , Transdução de Sinais , Regulação para CimaRESUMO
AIM: To determine the sub-macular Bruch's membrane (BrM) macrophage count and the choroidal and BrM macrophage immunophenotype in normal eyes and in eyes with early and advanced age-related macular degeneration (AMD). METHODS: BrM macrophages were counted in 125 human eyes (normal, normal aged, early AMD and geographical atrophy), and CD68 and inducible nitric oxide synthase (iNOS) immunohistochemistry was performed on 16 human eyes (normal, normal aged, early AMD, geographical atrophy and disciform scarring). All eyes were examined clinically ante mortem. Results were correlated with histopathological features, including basal laminar deposit and membranous debris, and with clinical fundus appearance. RESULTS: CD68(+) macrophages were found in the choroid of normal human eyes, and did not express iNOS. Expression of iNOS by choroidal macrophages (as well as endothelial cells and pericytes) was associated with: (1) recruitment of macrophages to BrM in early AMD eyes with soft drusen or thick continuous basal laminar deposit, corresponding to clinically detectable soft drusen or pigment changes; and (2) active disciform scarring. iNOS expression was absent in BrM macrophages, suggesting immunomodulatory differences between the choroid and BrM. The highest BrM macrophage counts were found in eyes with subclinical choroidal neovascularisation. CONCLUSION: The presence of extracellular deposits (soft drusen and thick continuous basal laminar deposit) is associated with macrophage recruitment to BrM and alteration in the immunophenotype of resident choroidal macrophages.