RESUMEN
Interphotoreceptor retinoid-binding protein (IRBP), a glycoprotein specific for the retina and pineal gland, induces inflammatory changes in these two organs in immunized animals. We report here on the identification of an immunodominant determinant of bovine IRBP that is highly immunogenic and immunopathogenic in the Lewis rat. The peptide, which comprises the sequence 1169-1191 of bovine IRBP, was shown to be immunodominant by its capacity to stimulate lymphocytes sensitized against whole IRBP. A comparison was made between peptide 1169-1191 and another peptide, 1158-1180, which is nondominant but is immunogenic and immunopathogenic in the Lewis rat. Peptide 1169-1191 was found to be superior in its immunological capacities; the minimal dose of 1169-1191 needed to induce cellular immune response or disease in Lewis rats (0.02-0.1 nmol/rat) is congruent to 1,000 times smaller than that of 1158-1180. In addition, unlike the ocular disease induced by 1158-1180, the disease produced by 1169-1191 resembled that induced by whole IRBP in its kinetics and histopathological features. The immunological activity of 1169-1191 in the Lewis rat was localized to the 10 residues at the COOH terminus; no such activity was exhibited by the truncated peptide 1169-1188, which comprises the 20 residues at the NH2 terminus of the full peptide. The usefulness of this unique experimental system in analyzing the role of immunodominance in peptide immunogenicity and immunopathogenicity is underscored.
Asunto(s)
Epítopos/análisis , Proteínas del Ojo/inmunología , Glándula Pineal , Proteínas de Unión al Retinol/inmunología , Uveítis/inmunología , Secuencia de Aminoácidos , Animales , Reacciones Cruzadas , Epítopos/administración & dosificación , Epítopos/inmunología , Inmunización Pasiva , Transfusión de Linfocitos , Linfocitos/inmunología , Masculino , Datos de Secuencia Molecular , Fragmentos de Péptidos/administración & dosificación , Fragmentos de Péptidos/síntesis química , Fragmentos de Péptidos/inmunología , Glándula Pineal/patología , Ratas , Ratas Endogámicas Lew , Uveítis/etiología , Uveítis/patologíaRESUMEN
Osteoclast differentiation factor (ODF, also called RANKL/TRANCE/OPGL) stimulates the differentiation of osteoclast progenitors of the monocyte/macrophage lineage into osteoclasts in the presence of macrophage colony-stimulating factor (M-CSF, also called CSF-1). When mouse bone marrow cells were cultured with M-CSF, M-CSF-dependent bone marrow macrophages (M-BMM phi) appeared within 3 d. Tartrate-resistant acid phosphatase-positive osteoclasts were also formed when M-BMM phi were further cultured for 3 d with mouse tumor necrosis factor alpha (TNF-alpha) in the presence of M-CSF. Osteoclast formation induced by TNF-alpha was inhibited by the addition of respective antibodies against TNF receptor 1 (TNFR1) or TNFR2, but not by osteoclastogenesis inhibitory factor (OCIF, also called OPG, a decoy receptor of ODF/RANKL), nor the Fab fragment of anti-RANK (ODF/RANKL receptor) antibody. Experiments using M-BMM phi prepared from TNFR1- or TNFR2-deficient mice showed that both TNFR1- and TNFR2-induced signals were important for osteoclast formation induced by TNF-alpha. Osteoclasts induced by TNF-alpha formed resorption pits on dentine slices only in the presence of IL-1alpha. These results demonstrate that TNF-alpha stimulates osteoclast differentiation in the presence of M-CSF through a mechanism independent of the ODF/RANKL-RANK system. TNF-alpha together with IL-1alpha may play an important role in bone resorption of inflammatory bone diseases.
Asunto(s)
Proteínas Portadoras/metabolismo , Glicoproteínas de Membrana/metabolismo , Osteoclastos/citología , Factor de Necrosis Tumoral alfa/metabolismo , Fosfatasa Ácida/metabolismo , Animales , Antígenos CD/genética , Antígenos CD/metabolismo , Células de la Médula Ósea/citología , Células de la Médula Ósea/efectos de los fármacos , Células de la Médula Ósea/metabolismo , Proteínas Portadoras/farmacología , Diferenciación Celular , Células Cultivadas , Expresión Génica , Humanos , Interleucina-1/metabolismo , Isoenzimas/metabolismo , Factor Estimulante de Colonias de Macrófagos/metabolismo , Factor Estimulante de Colonias de Macrófagos/farmacología , Masculino , Glicoproteínas de Membrana/farmacología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Osteoclastos/efectos de los fármacos , Osteoclastos/metabolismo , Osteoclastos/fisiología , Ligando RANK , Receptor Activador del Factor Nuclear kappa-B , Receptores del Factor de Necrosis Tumoral/genética , Receptores del Factor de Necrosis Tumoral/metabolismo , Receptores Tipo I de Factores de Necrosis Tumoral , Receptores Tipo II del Factor de Necrosis Tumoral , Fosfatasa Ácida Tartratorresistente , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
OBJECTIVE: Our goal was to evaluate the associations of antibodies (Abs) to glucose-6-phosphate isomerase (GPI) with Abs to cyclic citrullinated peptide (CCP) and HLA-DRB1 genotypes in Japanese patients with early rheumatoid arthritis (RA). METHODS: One hundred and eight patients with early RA (85 female, 23 male) who visited our clinic within 1 year of symptom onset were examined for anti-GPI and anti-CCP Ab levels, and HLA-DRB1 genotype. Anti-GPI and anti-CCP Ab levels, and HLA-DRB1 genotypes were also determined in 63 controls and 265 healthy controls, respectively. RESULTS: Of the 108 patients with early RA and the 63 controls, 20 (18.5%) and 3 (4.8%) were anti-GPI Ab-positive, respectively. Of the 20 patients with anti-GPI Abs, 17 (85%) were positive for anti-CCP Abs. HLA-DRB1*0405 and shared epitope (SE) carrier frequencies were significantly increased not only in anti-GPI Ab-positive patients (p=0.00057, odds ratio [OR] 4.6, 95% CI 1.8-11.8; p=0.0011, OR 5.0, 95% CI 1.7-14.0), but also in anti-GPI Ab-negative patients (p=0.0017, OR 2.2, 95% CI 1.3-3.7; p=0.00011, OR 2.6, 95% CI 1.6-4.3), when compared with controls. In addition, the carrier frequency of HLA-DRB1*1201 was significantly increased in anti-GPI Ab-positive patients compared with controls (p=0.0056, OR 4.3, 95% CI 1.4-13.2). CONCLUSIONS: The majority of anti-GPI Ab-positive RA patients constitute a subset of HLA-DRB1* SE-associated, anti-CCP Ab-positive RA patients.
Asunto(s)
Artritis Reumatoide/genética , Predisposición Genética a la Enfermedad/genética , Glucosa-6-Fosfato Isomerasa/inmunología , Antígenos HLA-DR/genética , Péptidos Cíclicos/inmunología , Adulto , Artritis Reumatoide/inmunología , Autoanticuerpos , Estudios de Casos y Controles , Femenino , Genotipo , Cadenas HLA-DRB1 , Humanos , Japón , Masculino , Persona de Mediana Edad , Oportunidad RelativaRESUMEN
IL-17 is a newly discovered T cell-derived cytokine whose role in osteoclast development has not been fully elucidated. Treatment of cocultures of mouse hemopoietic cells and primary osteoblasts with recombinant human IL-17 induced the formation of multinucleated cells, which satisfied major criteria of osteoclasts, including tartrate-resistant acid phosphatase activity, calcitonin receptors, and pit formation on dentine slices. Direct interaction between osteoclast progenitors and osteoblasts was required for IL-17-induced osteoclastogenesis, which was completely inhibited by adding indomethacin or NS398, a selective inhibitor of cyclooxgenase-2 (COX-2). Adding IL-17 increased prostaglandin E2 (PGE2) synthesis in cocultures of bone marrow cells and osteoblasts and in single cultures of osteoblasts, but not in single cultures of bone marrow cells. In addition, IL-17 dose-dependently induced expression of osteoclast differentiation factor (ODF) mRNA in osteoblasts. ODF is a membrane-associated protein that transduces an essential signal(s) to osteoclast progenitors for differentiation into osteoclasts. Osteoclastogenesis inhibitory factor (OCIF), a decoy receptor of ODF, completely inhibited IL-17-induced osteoclast differentiation in the cocultures. Levels of IL-17 in synovial fluids were significantly higher in rheumatoid arthritis (RA) patients than osteoarthritis (OA) patients. Anti-IL-17 antibody significantly inhibited osteoclast formation induced by culture media of RA synovial tissues. These findings suggest that IL-17 first acts on osteoblasts, which stimulates both COX-2-dependent PGE2 synthesis and ODF gene expression, which in turn induce differentiation of osteoclast progenitors into mature osteoclasts, and that IL-17 is a crucial cytokine for osteoclastic bone resorption in RA patients.
Asunto(s)
Artritis Reumatoide/metabolismo , Interleucina-17/fisiología , Osteoclastos/patología , Líquido Sinovial/metabolismo , Animales , Antígenos CD/inmunología , Artritis Reumatoide/inmunología , Células de la Médula Ósea/citología , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , División Celular/efectos de los fármacos , División Celular/fisiología , Técnicas de Cocultivo , Inhibidores de la Ciclooxigenasa/farmacología , Dinoprostona/metabolismo , Regulación de la Expresión Génica/fisiología , Humanos , Indometacina/farmacología , Interleucina-17/metabolismo , Masculino , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , Ratones , Nitrobencenos/farmacología , Osteoclastos/efectos de los fármacos , Ligando RANK , Receptor Activador del Factor Nuclear kappa-B , Sulfonamidas/farmacologíaRESUMEN
We describe two cases of Behçet's disease in a 37-year-old woman and a 40-year-old woman. Each of these patients developed cryptogenic organizing pneumonia associated with Behçet's disease. Both patients developed fever, cough and pleuritic chest pain during follow-up by our out-patient clinic. Chest X-rays and computed tomographies developed ground glass opacity, peripheral nodular opacities and consolidations. There were neither thrombosis nor aneurysm findings. Both cases were diagnosed as having organizing pneumonia. Prednisolone (40-60 mg/day) showed clinical and radiological improvement for both cases. Lung involvement is a rare feature of Behçet's disease, sometimes leading to a poor prognosis. The pulmonary parenchymal involvement still needs to be explored fully to diagnose early and start proper treatment. These two rare cases provide clinical insight into lung involvement in Behçet's disease.
Asunto(s)
Síndrome de Behçet/complicaciones , Neumonía en Organización Criptogénica/etiología , Neumonía en Organización Criptogénica/patología , Adulto , Antiinflamatorios/uso terapéutico , Síndrome de Behçet/tratamiento farmacológico , Neumonía en Organización Criptogénica/tratamiento farmacológico , Femenino , Humanos , Prednisolona/uso terapéuticoRESUMEN
OBJECTIVE: To evaluate the role of HLA-DRB1 genotypes and antibodies to cyclic citrullinated peptides (anti-CCP antibodies) in the development and radiographic progression of Japanese patients with rheumatoid arthritis (RA). METHODS: One hundred and ten patients with early RA (88 female, 22 male) who visited our clinic within 1 year of symptom onset were examined for anti-CCP antibody levels and HLA-DRB1 genotypes. HLA-DRB1 genotypes were also determined in 265 healthy controls. Radiographic progression over a 2-year interval was evaluated using the Larsen's method in 66 patients. RESULTS: Among the 110 patients with early RA, 82 patients (74.5%) were anti-CCP positive. Carrier frequency of HLA-DRB1*0405 was significantly increased in RA patients with anti-CCP antibodies compared with controls and RA patients without anti-CCP antibodies (odds ratio [OR] 3.4, 95% confidence interval [95% CI] 2.0-5.7 and OR 3.3, 95% CI 1.3-8.6, respectively). Carriership of one or two SE alleles was significantly associated with production of anti-CCP antibodies (OR 2.7, 95% CI 1.1-6.7 and OR 9.3, 95% CI 1.1-78.2, respectively). On the other hand, allele frequency of HLA-DRB1*0901 was significantly increased in RA patients without anti-CCP antibodies compared with controls and RA patients with anti-CCP antibodies (OR 2.2, 95% CI 1.1-4.1 and OR 3.0, 95% CI 1.4-6.4, respectively). CONCLUSION: In Japanese patients with RA, HLA-DRB1 SE alleles are associated with production of anti-CCP antibodies and HLA-DRB1 alleles appear to be differently associated with early RA depending on anti-CCP positivity as in Caucasian patients with RA.
Asunto(s)
Alelos , Artritis Reumatoide/genética , Artritis Reumatoide/inmunología , Autoanticuerpos/sangre , Antígenos HLA-DR/genética , Péptidos Cíclicos/inmunología , Adulto , Artritis Reumatoide/etnología , Pueblo Asiatico/etnología , Pueblo Asiatico/genética , Estudios de Casos y Controles , Progresión de la Enfermedad , Epítopos/genética , Femenino , Pie/diagnóstico por imagen , Genotipo , Cadenas HLA-DRB1 , Mano/diagnóstico por imagen , Heterocigoto , Humanos , Japón , Masculino , Persona de Mediana Edad , Péptidos Cíclicos/genética , RadiografíaRESUMEN
We describe two cases, a 28-year-old woman and a 46-year-old man, with mouth and genital ulcers with inflamed cartilage (chondritis of the nose and ears) (MAGIC syndrome). The conditions of both patients were resolved by treatment with corticosteroid and colchicine. We also review the English literature related to this rare syndrome.
Asunto(s)
Síndrome de Behçet/patología , Policondritis Recurrente/patología , Adulto , Síndrome de Behçet/complicaciones , Cartílago/patología , Femenino , Enfermedades de los Genitales Femeninos , Enfermedades de los Genitales Masculinos , Humanos , Masculino , Persona de Mediana Edad , Úlceras Bucales , Policondritis Recurrente/complicaciones , SíndromeRESUMEN
OBJECTIVES: Inflammatory mediators such as interleukin-6 and tumor necrosis factor-alpha play an important role in the pathogenesis of rheumatoid arthritis (RA) by promoting chronic inflammation and joint damage. NF-kappaB is a transcriptional activator of genes for these cytokines. It also plays an important role in the regulation of osteoclast differentiation which plays a key role in joint destruction in RA. Ligands for peroxisome proliferator-activated receptor (PPAR) -gamma have recently been reported to inhibit the development of RA. In this study, we investigated the role of PPARalpha in RA. METHODS: We analyzed the protein expression of PPAR-alpha and -gamma in rheumatoid synovial fibroblasts (RSF) from RA patients and analyzed the effects of ligands for PPAR-alpha and -gamma on cytokine production from RSF NF-kappaB activations in RSF and osteoclast differentiation from osteocalst progenitor in the peripheral blood. Moreover, we analyzed the effects of oral administration of PPAR-alpha and -gamma ligands on the development of adjuvant-induced arthritis (AIA) in female Lewis rats. RESULTS: We confirmed the expression of PPAR-alpha in RSF and also demonstrated that fenofibrate, a ligand for PPAR-alpha, inhibited cytokine production from RSF, NF- kappaB activation in RSF, and osteoclast differentiation from osteoclast progenitor cells. Furthermore, we demonstrated that fenofibrate inhibits the development of arthritis in a rat model of human RA. CONCLUSIONS: These results indicate that fenofibrate suppresses the development of arthritis by inhibition of NF-kappaB signaling; therefore, this compound offers possible anti-rheumatic drug.
Asunto(s)
Antirreumáticos , Artritis Reumatoide/metabolismo , Fenofibrato/farmacología , Hipolipemiantes/farmacología , FN-kappa B/antagonistas & inhibidores , PPAR alfa/metabolismo , Animales , Antirreumáticos/farmacología , Antirreumáticos/uso terapéutico , Artritis Experimental/tratamiento farmacológico , Artritis Experimental/etiología , Artritis Experimental/patología , Artritis Reumatoide/tratamiento farmacológico , Células Cultivadas , Relación Dosis-Respuesta a Droga , Femenino , Fenofibrato/uso terapéutico , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Fibroblastos/patología , Humanos , Hipolipemiantes/uso terapéutico , Ligandos , Osteoclastos/efectos de los fármacos , Osteoclastos/metabolismo , Osteoclastos/patología , PPAR gamma/metabolismo , Ratas , Ratas Endogámicas Lew , Transducción de Señal/efectos de los fármacos , Membrana Sinovial/efectos de los fármacos , Membrana Sinovial/metabolismo , Membrana Sinovial/patologíaRESUMEN
Osteopetrosis, a skeletal disorder of inadequate bone resorption with an abnormal increase in skeletal mass, results from a variety of independent single gene mutations that affect osteoclast differentiation and/or function. The osteopetrotic defect, op, is one of four spontaneous, nonallelic mutations in rats that result in osteopetrosis. In intercross progeny of (BN/SsN x LEW/SsN. +/op) F1 carriers, we mapped this locus by linkage analysis with microsatellite markers to rat chromosome 10. The linkage group contained, as well as op, 15 anonymous DNA loci and 9 DNA loci associated with genes (interleukin-3, myosin heavy chain [skeletal, embryonic], asialoglycoprotein receptor [hepatic lectin]-1, vesicle-associated membrane protein [synaptobrevin-2], sex hormone binding globulin, aldolase C, nitric oxide synthase [inducible], erythroblastic leukemia avian viral oncogene homolog-2, and proline-rich protein). The markers for these loci include nine not previously reported. The op locus mapped to the end of the chromosome 10 linkage group, within 1 cM of the anonymous DNA locus, D10Mit6. Based on its location, the op gene is likely to be distinct from seven described mutations in mice as well as three other mutations in rats. These results may permit a positional cloning strategy to be undertaken to identify the gene and mutation underlying the op defect.
Asunto(s)
Mapeo Cromosómico , Ligamiento Genético , Osteopetrosis/genética , Animales , Femenino , Código Genético , Marcadores Genéticos , Genotipo , Masculino , Meiosis/genética , Datos de Secuencia Molecular , Mutación , Ratas , Ratas Endogámicas LewRESUMEN
Chronic immune responses and inflammatory reactions in rheumatoid arthritis (RA) often cause severe destruction of cartilage and bone, but its mechanism is still a matter of controversy. We reported that interleukin-6 (IL-6) alone does not induce osteoclast formation, but soluble interleukin-6 receptors (sIL-6R) triggered the formation in the presence of IL-6 in cocultures of murine osteoblastic cells and bone marrow cells. In this study, we examined the involvement of sIL-6R and IL-6 in joint destruction in patients with RA. Although the frequency of patients having osteoclast-like multinucleated cells in synovium derived from the knee joint was not significantly different between RA (65%) and osteoarthritis (OA) patients (43%), the number of osteoclast-like cells found in the synovium was greater in the former than in the latter. Multinucleated cells obtained from RA synovium expressed the osteoclast-specific phenotype such as tartrate-resistant acid phosphatase, carbonic anhydrase II, vacuolar proton-ATPase and vitronectin receptors at similar levels to those from a human giant cell tumor of bone. The concentration of both IL-6 and sIL-6R was significantly higher in the synovial fluids from patients with RA than with OA. The concentration of IL-6 and sIL-6R correlated well with the roentgenologic grades of joint destruction. Dose-response curves for human IL-6 and human sIL-6R in inducing osteoclast-like cell formation in cocultures indicated that the RA synovial fluids contained sufficient IL-6 and sIL-6R to induce osteoclastogenesis. When synovial fluids from RA and OA patients were added to the cocultures, some of the RA synovial fluids containing high levels of IL-6 and sIL-6R stimulated osteoclast-like cell formation, which was strikingly inhibited by adding anti-IL-6R antibody simultaneously. These results suggest that IL-6 in the RA synovial fluids is at least in part responsible for joint destruction in the presence of sIL-6R through osteoclastogenesis.
Asunto(s)
Antígenos CD/metabolismo , Artritis Reumatoide/inmunología , Artritis Reumatoide/patología , Interleucina-6/metabolismo , Osteoclastos/inmunología , Osteoclastos/patología , Receptores de Interleucina/metabolismo , Líquido Sinovial/inmunología , Fosfatasa Ácida/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Animales , Artritis Reumatoide/etiología , Células Cultivadas , Femenino , Humanos , Masculino , Ratones , Persona de Mediana Edad , Osteoartritis/inmunología , Osteoartritis/patología , Osteoblastos/patología , Osteoclastos/enzimología , Fenotipo , Receptores de Interleucina-6 , Solubilidad , Membrana Sinovial/enzimología , Membrana Sinovial/inmunología , Membrana Sinovial/patologíaRESUMEN
We have extended the cDNA sequence of bovine interphotoreceptor retinoid-binding protein (IRBP) and subcloned one of the sequenced cDNA fragments into an expression vector. The nucleotide (nt) sequences of four bovine IRBP cDNA clones have been determined. These sequences when assembled cover the 3' proximal 3629 nt of the IRBP mRNA and encode the C-terminal 551 amino acids (aa) of IRBP. This cDNA sequence validates the intron: exon boundaries predicted from the gene. A 2-kb EcoRI insert from lambda IRBP2, one of the clones sequenced, encoding the C-terminal 136 aa of IRBP was subcloned into the expression vector pWR590-1. Escherichia coli carrying this plasmid construction, pXS590-IRBP, produced a fusion protein containing 583 N-terminal aa of beta-galactosidase, three linker aa residues, 136 C-terminal aa of IRBP and possibly a number of additional C-terminal residues due to suppressed termination. This 86-kDa fusion protein, purified by detergent/chaotrope extraction followed by reverse-phase high-performance liquid chromatography, cross-reacted with anti-bovine IRBP on Western blots. This protein induced an experimental autoimmune uveo-retinitis and experimental autoimmune pinealitis in Lewis rats indistinguishable from that induced by authentic bovine IRBP. Thus, it is evident that biological activity of this region of IRBP, as manifested by immuno-pathogenicity, is retained by the fusion protein.
Asunto(s)
Proteínas del Ojo/genética , Proteínas Recombinantes de Fusión/genética , Proteínas de Unión al Retinol/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Bovinos , Cromatografía Líquida de Alta Presión , Clonación Molecular , ADN/genética , Escherichia coli/genética , Ojo/inmunología , Ojo/patología , Oftalmopatías/etiología , Oftalmopatías/inmunología , Oftalmopatías/patología , Proteínas del Ojo/inmunología , Masculino , Datos de Secuencia Molecular , Plásmidos , Ratas , Proteínas Recombinantes de Fusión/inmunología , Proteínas de Unión al Retinol/inmunologíaRESUMEN
It is difficult to quantitate cytokine mRNA profiles in small human tissue specimens obtained by a needle biopsy, even using standard RT-PCR methods, because the amount of mRNA in the specimens is very small. To address this problem, we developed highly sensitive, quantitative, nested RT-PCR techniques to evaluate the expression of multiple cytokine mRNAs in synovial specimens obtained by needle biopsy. To reduce effects of variation of initial RNA concentrations, cDNA from each target RNA sample was normalized, using a simplified competitive PCR method, to the levels of beta-actin cDNA. The first and the second (nested) PCR were performed in the same tube to prevent contamination. The number of PCR-product bands, evident on polyacrylamide gel electrophoresis, was used to quantitate the relative amounts of target cDNA. Using our methods, it was possible to evaluate, in a single synovial tissue specimen obtained by needle biopsy, the relative amounts of mRNAs for 10 cytokines (TNF-alpha, IL-1 beta, IL-2, IL-4, IL-6, IL-10, IL-12 p40, IL-13, IL-15, IFN-gamma) and CD3 delta chain. Our methods are particularly valuable if there are multiple target mRNAs, numerous samples, or if the amounts of mRNAs are limited. The methods are applicable to a wide variety of tissues and target mRNAs.
Asunto(s)
Citocinas/genética , Reacción en Cadena de la Polimerasa/métodos , ARN Mensajero/análisis , Artritis Reumatoide/fisiopatología , Biopsia con Aguja/métodos , Cartilla de ADN , Expresión Génica , Humanos , Rodilla , ARN Mensajero/genética , Líquido Sinovial/químicaRESUMEN
Peptide R23, consisting of residues 1091-1115 of bovine interphotoreceptor retinoid-binding protein (IRBP), had some unusual immunologic properties in Lewis rats. The peptide induced experimental autoimmune uveoretinitis and pinealitis in these rats, but only at high doses. The minimal immunopathogenic dose was found to be 100 nmol/rat. On the other hand, R23 was highly immunogenic in Lewis rats, producing cellular immunity, as measured by the lymphocyte proliferation assay, with a minimal dose of 1 nmol/rat. This unusual dissociation between the uveitogenic and immunogenic activities of R23 was attributed to different sites on the peptide, stimulating either the lymphocytes which induce disease or those which vigorously proliferate in culture. The potent immunogenicity of R23 was consistent with the peptide being immunodominant, as demonstrated by its capacity to be recognized by lymphocytes sensitized against whole IRBP and to stimulate these cells in culture to proliferate and acquire uveitogenic capacity. Likewise, lymphocytes sensitized against R23 are stimulated in culture by whole IRBP. Unexpectedly, peptide R23 was inferior to whole IRBP in its capacity to stimulate uveitogenicity in R23-sensitized lymphocytes. This finding also was attributed to the preferential stimulation by R23 of the lymphocytes specific for the putative "nonuveitogenic" site on the peptide. Peptide R23 also differs from the other tested bovine IRBP-derived peptides in the specificity of its antibodies. Unlike antibodies to the other peptides, those to R23 showed a strong cross reactivity toward whole IRBP.
Asunto(s)
Proteínas del Ojo/inmunología , Proteínas de Unión al Retinol/inmunología , Uveítis/inmunología , Secuencia de Aminoácidos , Animales , Autoanticuerpos/análisis , Reacciones Cruzadas , Inmunidad Celular , Epítopos Inmunodominantes , Inmunoterapia Adoptiva , Activación de Linfocitos , Masculino , Datos de Secuencia Molecular , Ratas , Ratas Endogámicas Lew , Uveítis/patologíaRESUMEN
PURPOSE: To investigate pathogenesis underlying endogenous uveitis, macrophage migration inhibitory factor (MIF) was quantified in sera of patients. METHODS: Sera were obtained from the 55 patients with uveitis (24 with Behçet's disease; 9 with Vogt-Koyanagi-Harada's [VKH] disease; 22 with sarcoidosis) and 58 healthy control subjects. MIF levels were determined by a human MIF enzyme-linked immunosorbent assay. RESULTS: The mean MIF levels in the sera of the patients with Behçet's disease, VKH disease, and sarcoidosis and of healthy control subjects were 60.4+/-9.0 (mean+/-SE) ng/ml, 16.5+/-2.9 ng/ml, 27.1+/-5.6 ng/ml, and 5.4+/-0.04 ng/ml, respectively. The average levels of MIF in the sera of uveitis patients were significantly higher (P < 0.0001) than those of healthy control subjects. The high levels of MIF were especially noted in patients with Behçet's disease at the ocular exacerbation stage and patients with sarcoidosis at the severe uveitis stage. CONCLUSIONS: Significant increase of MIF in sera was characteristic of uveitis, and MIF may be a usefull laboratory parameter to use to comprehend the clinical course of uveitis.
Asunto(s)
Factores Inhibidores de la Migración de Macrófagos/sangre , Uveítis/sangre , Síndrome de Behçet/sangre , Ensayo de Inmunoadsorción Enzimática , Oftalmopatías/sangre , Humanos , Sarcoidosis/sangre , Síndrome Uveomeningoencefálico/sangreRESUMEN
PURPOSE: HLA-B27-associated acute anterior uveitis (HLA-B27 AAU) seems to be triggered by external factors in persons with a particular genetic background. It is still uncertain whether HLA-B27 or other gene(s) near the HLA-B region predisposes to uveitis in a linkage disequilibrium with B27. The authors investigated microsatellite polymorphism within the transmembrane region of the MICA gene, located 47 kb centromeric of the HLA-B gene, and HLA-B27 subtypes. METHODS: Seventeen HLA-B27-positive Japanese patients with HLA-B27 AAU, 51 Japanese controls, and 20 B27-positive Japanese controls were examined for MICA gene polymorphism within the transmembrane region using polymerase chain reaction (PCR) and subsequent automated fragment detection by fluorescent-based technology. Furthermore, B27-positive patients with HLA-B27 AAU and B27-positive controls were examined for HLA-B27 subtypes by the PCR-sequence-specific primer method. RESULTS: The microsatellite allele in the MICA gene, consisting of four repetitions of GCT/AGC (designated A4 allele), was present at a significantly higher phenotype frequency in the patient group (64.7%) than in the control group (25.5%) (chi 2 = 6.95, Pc = 0.042). Furthermore, the frequency of the A4 allele was significantly higher, even when compared with 20% in the B27-positive control group (chi 2 = 5.88, Pc = 0.042). The frequency of HLA-B27 subtypes was not significantly different between B27-positive patients with HLA-B27 AAU and B27-positive controls. CONCLUSIONS: These results suggest that the MICA gene itself, or other nearby gene(s), linked to the MICA A4 allele may be involved in the development of HLA-B27 AAU and that HLA-B27 subtypes are not important in the development of HLA-B27 AAU in a Japanese population.
Asunto(s)
Antígeno HLA-B27/genética , Antígenos de Histocompatibilidad Clase I/genética , Polimorfismo Genético , Uveítis Anterior/genética , Enfermedad Aguda , Adulto , Alelos , Cartilla de ADN/química , ADN Satélite/análisis , Electroforesis en Gel de Agar , Femenino , Frecuencia de los Genes , Genotipo , Humanos , Masculino , Reacción en Cadena de la PolimerasaRESUMEN
To elucidate the role of macrophage migration inhibitory factor (MIF) in ocular inflammation, we examined the localization of MIF in the normal anterior uveal tract of rats. Immunohistochemistry using an anti-MIF antibody revealed that MIF was present in non-pigment epithelial cells of the ciliary body and the epithelial cells of the iris. Western blot analysis of these tissues showed a single band specific for MIF protein. The expression of MIF mRNA in these tissues was further confirmed by reverse transcription-polymerase chain reaction. Since MIF is known to be a potent proinflammatory cytokine, identification of the protein in iris and ciliary epithelial cells suggests the possibility that it may play an important role in ocular inflammation.
Asunto(s)
Cuerpo Ciliar/inmunología , Iris/inmunología , Factores Inhibidores de la Migración de Macrófagos/inmunología , Animales , Western Blotting , Epitelio , Técnicas para Inmunoenzimas , Factores Inhibidores de la Migración de Macrófagos/genética , Reacción en Cadena de la Polimerasa , Conejos , Ratas , Ratas Wistar , Transcripción GenéticaRESUMEN
It has been shown that macrophage migration inhibitory factor (MIF) modulates not only macrophage functions, but also T cell functions. However, detailed analysis of the MIF function on responses of various T cell subpopulations remained to be elucidated. In this report, using a neutralizing anti-MIF monoclonal antibody (mAb) we examined MIF functions on various T cell lineages. It was shown that anti-MIF mAb inhibited antigen-specific responses of both IFN-gamma producing and IL-4 producing T cells. The inhibition appeared to be related to blockade of the signal pathway via T cell receptor (TCR) but not that via IL-2 receptor (IL-2R). However, the anti-MIF mAb showed no inhibitory effect on NK-T cell responses stimulated through TCR. These results suggest that MIF is involved in the signal pathway via TCR in mainstream T cells but not in NK-T cells.
Asunto(s)
Factores Inhibidores de la Migración de Macrófagos/inmunología , Transducción de Señal/inmunología , Subgrupos de Linfocitos T/inmunología , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales/inmunología , Especificidad de Anticuerpos/inmunología , Antígenos/inmunología , Antígenos de Superficie , División Celular , Línea Celular , Humanos , Células Asesinas Naturales/citología , Células Asesinas Naturales/metabolismo , Lectinas Tipo C , Activación de Linfocitos , Factores Inhibidores de la Migración de Macrófagos/biosíntesis , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Subfamilia B de Receptores Similares a Lectina de Células NK , Proteínas/inmunología , Receptores de Antígenos de Linfocitos T/inmunología , Receptores de Interleucina-2/inmunologíaRESUMEN
We determined the antibody response against Borrelia burgdorferi strains isolated from Japanese Ixodes ovatus and Ixodes persulcatus ticks by enzyme-linked immunosorbent assay and indirect immunofluorescence assay of serum specimens from 127 patients with uveitis. We examined samples of serum from Japanese patients with unclassified uveitis, iridocyclitis caused by herpes zoster virus, Behçet's disease, Vogt-Koyanagi-Harada syndrome, sarcoidosis, or other conditions (sympathetic ophthalmia, Posner-Schlossman syndrome and acute anterior uveitis with ankylosing spondylitis). Serum from healthy individuals and patients with Lyme disease served as negative and positive control samples, respectively. Significantly higher antibody titers were demonstrated in patients with uveitis than in control subjects. Of 29 patients with unclassified uveitis, nine (31) had significantly increased antibody titers against B. burgdorferi strain H014 by ELISA testing. Five patients also showed higher IgG and IgM responses than in three control subjects with Lyme disease. All positive controls showed joint problems characteristic of rheumatoid arthritis. One of three patients had uveitis. The patients were diagnosed as having Lyme disease on the basis of their history and serologic tests. A positive antibody response was recognized in several patients with Behçet's disease, Vogt-Koyanagi-Harada syndrome, sarcoidosis, and other conditions (acute anterior uveitis with ankylosing spondylitis), but not in control subjects.
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Anticuerpos Antibacterianos/análisis , Grupo Borrelia Burgdorferi/inmunología , Uveítis Anterior/inmunología , Uveítis Posterior/inmunología , Ensayo de Inmunoadsorción Enzimática , Técnica del Anticuerpo Fluorescente , Humanos , Inmunoglobulina G/inmunología , Inmunoglobulina M/inmunología , Enfermedad de Lyme/inmunología , Sarcoidosis/inmunología , Síndrome Uveomeningoencefálico/inmunologíaRESUMEN
AIMS: To determine whether macrophage migration inhibitory factor (MIF) levels were increased in sera of the patients with iridocyclitis. METHODS: Sera were obtained from 41 patients with acute iridocyclitis, 13 patients with chronic iridocyclitis, and 44 healthy control subjects. MIF levels were determined by a human MIF ELISA. RESULTS: The average levels of MIF in the sera of patients with both acute and chronic iridocyclitis were significantly higher than that of healthy subjects. CONCLUSION: Uveitis induces the elevation of serum MIF, which may affect various inflammatory symptoms in uveitis.
Asunto(s)
Iridociclitis/sangre , Factores Inhibidores de la Migración de Macrófagos/sangre , Enfermedad Aguda , Enfermedad Crónica , Femenino , Humanos , MasculinoRESUMEN
Specimens of synovial tissues from 5 affected joints of 3 patients with Behçet's disease were available for histopathological examination. All specimens were infiltrated by lymphocytes and neutrophils, and exhibited marked vascularity and infiltration of lymphoid cells among the vessels. Marked plasma cell infiltration and lymphoid follicle formation were found in one synovial tissue sample. There was no evidence of infection or vasculitis. These findings suggest that the histopathological characteristics of synovial tissue in Behçet's disease may have a wide range, some of which may even resemble the synovial tissue of rheumatoid arthritis.