RESUMEN
The bovine viral diarrhea virus (BVDV-1) is a pathogen with the capacity to modulate the interferon type I system. To further investigate the effects of BVDV-1 on the production of the immune response, the Madin-Darby bovine kidney cell line was infected with the cytopathic CH001 field isolate of BVDV-1, and the IFNbeta expression profiles were analyzed. The results showed that cpBVDV-1 was able to induce the production of IFNbeta in a way similar to polyinosinic-polycytidylic acid, but with less intensity. Interestingly, all cpBVDV-1 activities were blocked by pharmacological inhibitors of the IRF-1, IRF-7, and NF-κB signaling pathway, and the level of IFNbeta decreased at the level of transcript and protein. These results, together with in silico analyses showing the presence of several regulatory consensus target motifs, suggest that cpBVDV-1 regulates IFNbeta expression in bovines through the activation of several key transcription factors. Collectively, the results suggest that during cpBVDV-1 infection, cross talk is evident between various signaling pathways involved in transcriptional activation of IFNbeta in cattle.
Asunto(s)
Diarrea Mucosa Bovina Viral/genética , Virus de la Diarrea Viral Bovina Tipo 1/inmunología , Regulación de la Expresión Génica/genética , Expresión Génica/genética , Factor 1 Regulador del Interferón/genética , Factor 7 Regulador del Interferón/genética , FN-kappa B/genética , Animales , Diarrea Mucosa Bovina Viral/virología , Bovinos , Línea Celular , Células Epiteliales/inmunología , Células Epiteliales/virología , Expresión Génica/inmunología , Regulación de la Expresión Génica/inmunología , Factor 1 Regulador del Interferón/inmunología , Factor 7 Regulador del Interferón/inmunología , FN-kappa B/inmunología , Transducción de Señal/genética , Transducción de Señal/inmunologíaRESUMEN
The role and regulators of extracellular vesicle (EV) secretion in hepatic ischemia/reperfusion (IR) injury have not been defined. Rab27a is a guanosine triphosphatase known to control EV release. Interferon regulatory factor 1 (IRF-1) is a transcription factor that plays an important role in liver IR and regulates certain guanosine triphosphatases. However, the relationships among IRF-1, Rab27a, and EV secretion are largely unknown. Here, we show induction of IRF-1 and Rab27a both in vitro in hypoxic hepatocytes and in vivo in warm IR and orthotopic liver transplantation livers. Interferon γ stimulation, IRF-1 transduction, or IR promoted Rab27a expression and EV secretion. Meanwhile, silencing of IRF-1 decreased Rab27a expression and EV secretion. Rab27a silencing decreased EV secretion and liver IR injury. Ten putative IRF-1 binding motifs in the 1,692-bp Rab27a promoter region were identified. Chromatin immunoprecipitation and electrophoretic mobility shift assay verified five functional IRF-1 binding motifs, which were confirmed by a Rab27a promoter luciferase assay. IR-induced EVs contained higher oxidized phospholipids (OxPL). OxPLs on the EV surface activated neutrophils through the toll-like receptor 4 pathway. OxPL-neutralizing E06 antibody blocked the effect of EVs and decreased liver IR injury. CONCLUSION: These findings provide a novel mechanism by which IRF-1 regulates Rab27a transcription and EV secretion, leading to OxPL activation of neutrophils and subsequent hepatic IR injury. (Hepatology 2018;67:1056-1070).
Asunto(s)
Vesículas Extracelulares/metabolismo , Factor 1 Regulador del Interferón/metabolismo , Hígado/patología , Daño por Reperfusión/metabolismo , Proteínas rab27 de Unión a GTP/metabolismo , Animales , Técnicas de Cultivo de Célula , Regulación de la Expresión Génica , Hepatocitos/metabolismo , Humanos , Hígado/metabolismo , Trasplante de Hígado , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones NoqueadosRESUMEN
A high percentage of type 2 diabetes mellitus (T2D) patients are also affected by dyslipidemia and chronic periodontitis (CP), but no studies have determined the gene expression in patients that are simultaneously affected by all three diseases. We investigated the systemic expression of immune-related genes in T2D, dyslipidemia, and CP patients. One hundred and fifty patients were separated into five groups containing 30 individuals each: (G1) poorly controlled T2D with dyslipidemia and CP; (G2) well-controlled T2D with dyslipidemia and CP; (G3) normoglycemic individuals with dyslipidemia and CP; (G4) healthy individuals with CP; (G5) systemic and periodontally healthy individuals. Blood analyses of lipid and glycemic profiles were carried out. The expression of genes, including IL10, JAK1, STAT3, SOCS3, IP10, ICAM1, IFNA, IFNG, STAT1, and IRF1, was investigated by RT-qPCR. Patients with dyslipidemia demonstrated statistically higher expression of the IL10 and IFNA genes, while IFNG, IP10, IRF1, JAK1, and STAT3 were lower in comparison with nondyslipidemic patients. Anti-inflammatory genes, such as IL10, positively correlated with parameters of glucose, lipid, and periodontal profiles, while proinflammatory genes, such as IFNG, were negatively correlated with these parameters. We conclude that dyslipidemia appears to be the primary disease that is associated with gene expression of immune-related genes, while parameters of T2D and CP were correlated with the expression of these important immune genes.
Asunto(s)
Periodontitis Crónica/metabolismo , Diabetes Mellitus Tipo 2/metabolismo , Dislipidemias/metabolismo , Adulto , Periodontitis Crónica/genética , Diabetes Mellitus Tipo 2/genética , Dislipidemias/genética , Femenino , Humanos , Inflamación/genética , Inflamación/metabolismo , Molécula 1 de Adhesión Intercelular/genética , Molécula 1 de Adhesión Intercelular/metabolismo , Factor 1 Regulador del Interferón/genética , Factor 1 Regulador del Interferón/metabolismo , Interleucina-10/genética , Interleucina-10/metabolismo , Janus Quinasa 1/genética , Janus Quinasa 1/metabolismo , Masculino , Persona de Mediana Edad , Receptores de Citocinas/genética , Receptores de Citocinas/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Transcripción STAT1/genética , Factor de Transcripción STAT1/metabolismo , Proteína 3 Supresora de la Señalización de Citocinas/genética , Proteína 3 Supresora de la Señalización de Citocinas/metabolismoRESUMEN
Brucella abortus is an intracellular pathogen capable of surviving inside of macrophages. The success of B. abortus as a chronic pathogen relies on its ability to orchestrate different strategies to evade the adaptive CD4+ T cell responses that it elicits. Previously, we demonstrated that B. abortus inhibits the IFN-γ-induced surface expression of MHC class II (MHC-II) molecules on human monocytes, and this phenomenon correlated with a reduction in antigen presentation. However, the molecular mechanisms, whereby B. abortus is able to down-regulate the expression of MHC-II, remained to be elucidated. In this study, we demonstrated that B. abortus infection inhibits the IFN-γ-induced transcription of MHC-II, transactivator (CIITA) and MHC-II genes. Accordingly, we observed that the synthesis of MHC-II proteins was also diminished. B. abortus was not only able to reduce the expression of mature MHC-II, but it also inhibited the expression of invariant chain (Ii)-associated immature MHC-II molecules. Outer membrane protein 19 (Omp19), a prototypical B. abortus lipoprotein, diminished the expression of MHC-II and CIITA transcripts to the same extent as B. abortus infection. IL-6 contributes to these down-regulatory phenomena. In addition, B. abortus and its lipoproteins, through IL-6 secretion, induced the transcription of the negative regulators of IFN-γ signaling, suppressor of cytokine signaling (SOCS)-1 and -3, without interfering with STAT1 activation. Yet, B. abortus lipoproteins via IL-6 inhibit the expression of IFN regulatory factor 1 (IRF-1), a critical regulatory transcription factor for CIITA induction. Overall, these results indicate that B. abortus inhibits the expression of MHC-II molecules at very early points in their synthesis and in this way, may prevent recognition by T cells establishing a chronic infection.
Asunto(s)
Brucella abortus/fisiología , Regulación hacia Abajo , Antígenos de Histocompatibilidad Clase II/metabolismo , Factor 1 Regulador del Interferón/metabolismo , Interleucina-6/metabolismo , Proteínas Nucleares/antagonistas & inhibidores , Transactivadores/antagonistas & inhibidores , Antígenos Bacterianos/inmunología , Proteínas de la Membrana Bacteriana Externa/inmunología , Brucelosis/inmunología , Brucelosis/microbiología , Brucelosis/patología , Catepsinas/metabolismo , Línea Celular , Antígenos HLA-DR/inmunología , Humanos , Interferón gamma/metabolismo , Espacio Intracelular/metabolismo , Lipoproteínas/inmunología , Lipoproteínas/metabolismo , Modelos Biológicos , Monocitos/microbiología , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Fosforilación , Factor de Transcripción STAT1/metabolismo , Proteína 1 Supresora de la Señalización de Citocinas/metabolismo , Proteína 3 Supresora de la Señalización de Citocinas/metabolismo , Transactivadores/genética , Transactivadores/metabolismo , Transcripción GenéticaRESUMEN
Interferon regulatory factor 1 (IRF1) is functionally diverse in the regulation of immune response and is considered to be an important candidate gene for studying disease susceptibility in mammals. In this paper, we characterized the whole sequence of the IRF1 gene in river buffalo (Bubalus bubalis) and compared genomic and the amino acid sequences between different species. The buffalo IRF1 gene was 7099 bp long and organized into 10 exons and nine introns. Its molecular structure showed exactly the same number of exons (10) and introns (nine) in bovids, mice, horses, humans, and chickens. However, rats did not have exon 5, but had the largest exon 4, which suggests that exon 5 was incorporated into exon 4. The coding and the amino acid sequences of the gene showed that identity varied from 73 to 99% at the coding sequence level and from 61 to 100% at the amino acid level when compared with other mammals and chickens. Comparative analysis of the gene sequence between two different buffalo breeds, Murrah and Mediterranean, revealed six potential SNPs that are primarily located in the 5' and 3'UTRs.
Asunto(s)
Búfalos/genética , Factor 1 Regulador del Interferón/genética , Animales , Cromosomas Artificiales Bacterianos , Exones , Especiación Genética , Intrones , Filogenia , Análisis de Secuencia de ADN/métodos , Análisis de Secuencia de ADN/veterinaria , Análisis de Secuencia de Proteína/métodos , Análisis de Secuencia de Proteína/veterinariaRESUMEN
The bovine viral diarrhea virus (BVDV-1) is a pathogen responsible for high economic losses in the cattle industry worldwide. This virus has the capacity to modulate the immune system of several higher vertebrates, but there is little information available on the cell infection mechanism. To further investigate the effects of BVDV-1 on the activation of the immune response, the Madin-Darby bovine kidney cell line was infected with the cytopathic CH001 field isolate of BVDV-1, and the proinflammatory and antiviral cytokine expression profiles were analyzed. The results showed that BVDV-1 was able to induce the production of BCL3, IL-1ß, IL-8, IL-15, IL-18, Mx-1, IRF-1, and IRF-7 in a way similar to polyinosinic-polycytidylic acid. Interestingly, all BVDV-1 activities were blocked by pharmacological inhibitors of the NF-κB signaling pathway. These results, together with in silico analyses showing the presence of several regulatory consensus target motifs, suggest that BVDV-1 regulates gene expression in bovines through the activation of several key transcription factors. Collectively, these data identified BVDV-1 as a viral regulator of immune marker expression, even from early infection. Additionally, this is the first report to find BVDV-1 modulating the activation of cytokine production and transcriptions factors mainly through the NF-κB pathway in vertebrates.
Asunto(s)
Virus de la Diarrea Viral Bovina/efectos de los fármacos , Células Epiteliales/efectos de los fármacos , Interleucinas/antagonistas & inhibidores , FN-kappa B/antagonistas & inhibidores , Nitrilos/farmacología , Sulfonas/farmacología , Animales , Proteínas del Linfoma 3 de Células B , Biomarcadores/metabolismo , Bovinos , Línea Celular , Virus de la Diarrea Viral Bovina/crecimiento & desarrollo , Virus de la Diarrea Viral Bovina/inmunología , Células Epiteliales/inmunología , Células Epiteliales/patología , Células Epiteliales/virología , Regulación de la Expresión Génica , Interacciones Huésped-Patógeno , Factor 1 Regulador del Interferón/genética , Factor 1 Regulador del Interferón/inmunología , Factor 7 Regulador del Interferón/genética , Factor 7 Regulador del Interferón/inmunología , Interleucinas/genética , Interleucinas/inmunología , Proteínas de Resistencia a Mixovirus/genética , Proteínas de Resistencia a Mixovirus/inmunología , FN-kappa B/genética , FN-kappa B/inmunología , Poli I-C/farmacología , Proteínas Proto-Oncogénicas/genética , Proteínas Proto-Oncogénicas/inmunología , Transducción de Señal , Factores de Transcripción/genética , Factores de Transcripción/inmunologíaRESUMEN
Macrophage tumoricidal activity relies, mainly, on the release of Tumor Necrosis Factor alpha (TNFα) and/or on reactive oxygen or nitrogen intermediates. In the present work, we investigated the cytotoxic activity of resident peritoneal macrophages against L929 fibrosarcoma cell line in vitro and in vivo. Resident macrophages lysed L929 cells in a mechanism independent of TNFα and cell-to-cell contact. The cytotoxic activity was largely dependent on nitric oxide (NO) release since treatment with L-NAME (NOS inhibitor) inhibited L929 cells killing. Macrophages from mice with targeted deletion of inducible NO synthase (iNOS) together with L929 cells produced less NO and displayed lower, but still significant, tumoricidal activity. Notably, NO production and tumor lysis were abolished in co-cultures with macrophages deficient in Interferon Regulatory Factor, IRF-1. Importantly, the in vitro findings were reproduced in vivo as IRF-1 deficient animals inoculated i.p with L929 cells were extremely susceptible to tumor growth and their macrophages did not produce NO, while WT mice killed L929 tumor cells and their macrophages produced high levels of NO. Our results indicate that IRF-1 is a master regulator of bi-directional interaction between macrophages and tumor cells. Overall, IRF-1 was essential for NO production by co-cultures and macrophage tumoricidal activity in vitro as well as for the control of tumor growth in vivo.
Asunto(s)
Comunicación Celular/inmunología , Fibrosarcoma/inmunología , Factor 1 Regulador del Interferón/inmunología , Macrófagos Peritoneales/inmunología , Óxido Nítrico/inmunología , Animales , Línea Celular Tumoral , Femenino , Fibrosarcoma/patología , Macrófagos Peritoneales/patología , Masculino , Ratones , Ratones NoqueadosRESUMEN
Brucella species can cause brucellosis, a zoonotic disease that causes serious livestock economic losses and represents a public health threat. The mechanism of virulence of Brucella spp. is not yet fully understood. Therefore, it is crucial to identify new molecules that serve as virulence factors to better understand this host-pathogen interplay. Here, we evaluated the role of the Brucella membrane fusogenic protein (Mfp) and outer membrane protein 19 (Omp19) in bacterial pathogenesis. In this study, we showed that B. abortus Δmfp::kan and Δomp19::kan deletion mutant strains have reduced persistence in vivo in C57BL/6 and interferon regulatory factor 1 (IRF-1) knockout (KO) mice. Additionally, 24 h after macrophage infection with a Δmfp::kan or Δomp19::kan strain expressing green fluorescent protein (GFP) approximately 80% or 65% of Brucella-containing vacuoles (BCVs) retained the late endosomal/lysosomal marker LAMP-1, respectively, whereas around 60% of BCVs containing wild-type S2308 were found in LAMP-1-negative compartments. B. abortus Δomp19::kan was attenuated in vivo but had a residual virulence in C57BL/6 and IRF-1 KO mice, whereas the Δmfp::kan strain had a lower virulence in these same mouse models. Furthermore, Δmfp::kan and Δomp19::kan strains were used as live vaccines. Challenge experiments revealed that in C57BL/6 and IRF-1 KO mice, the Δmfp::kan strain induced greater protection than the vaccine RB51 and protection similar that of vaccine S19. However, a Δomp19::kan strain induced protection similar to that of RB51. Thus, these results demonstrate that Brucella Mfp and Omp19 are critical for full bacterial virulence and that the Δmfp::kan mutant may serve as a potential vaccine candidate in future studies.
Asunto(s)
Antígenos Bacterianos/genética , Proteínas de la Membrana Bacteriana Externa/genética , Brucella abortus/inmunología , Brucella abortus/patogenicidad , Brucelosis/inmunología , Lipoproteínas/genética , Proteínas de la Fusión de la Membrana/genética , Factores de Virulencia/genética , Animales , Antígenos Bacterianos/inmunología , Proteínas de la Membrana Bacteriana Externa/inmunología , Vacuna contra la Brucelosis/inmunología , Brucella abortus/genética , Brucelosis/patología , Brucelosis/prevención & control , Eliminación de Gen , Proteínas Fluorescentes Verdes/biosíntesis , Factor 1 Regulador del Interferón/genética , Lipoproteínas/inmunología , Proteínas de Membrana de los Lisosomas/metabolismo , Macrófagos/inmunología , Macrófagos/microbiología , Proteínas de la Fusión de la Membrana/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Vacunación , Factores de Virulencia/inmunologíaRESUMEN
UNLABELLED: Plasmacytoid dendritic cells (pDC) constitute the body's principal source of type I interferon (IFN) and are comparatively abundant in the liver. Among various cytokines implicated in liver ischemia and reperfusion (I/R) injury, type I IFNs have been described recently as playing an essential role in its pathogenesis. Moreover, type I IFNs have been shown to up-regulate hepatocyte expression of IFN regulatory factor 1 (IRF-1), a key transcription factor that regulates apoptosis and induces liver damage after I/R. Our aim was to ascertain the capacity of IFN-α released by liver pDC to induce liver damage through hepatic IRF-1 up-regulation after I/R injury. Our findings show that liver pDC mature and produce IFN-α in response to liver I/R. Liver pDC isolated after I/R induced elevated levels of IRF-1 production by hepatocytes compared with liver pDC isolated from sham-operated mice. Notably, hepatic IRF-1 expression was reduced significantly by neutralizing IFN-α. In vivo, IFN-α neutralization protected the liver from I/R injury by reducing hepatocyte apoptosis. This was associated with impaired expression of IRF-1 and proapoptotic molecules such as Fas ligand, its receptor (Fas) and death receptor 5, which are regulated by IRF-1. Furthermore, pDC-depleted mice failed to up-regulate hepatic IFN-α and displayed less liver injury associated with reduced levels of hepatic interleukin (IL)-6, tumor necrosis factor-α, and hepatocyte apoptosis after I/R compared with controls. CONCLUSION: these data support the hypothesis that IFN-α derived from liver pDC plays a key role in the pathogenesis of liver I/R injury by enhancing apoptosis as a consequence of induction of hepatocyte IRF-1 expression.
Asunto(s)
Células Dendríticas/inmunología , Factor 1 Regulador del Interferón/inmunología , Interferón-alfa/inmunología , Hepatopatías/inmunología , Daño por Reperfusión/inmunología , Animales , Apoptosis/inmunología , Células Cultivadas , Células Dendríticas/metabolismo , Modelos Animales de Enfermedad , Hepatocitos/citología , Hepatocitos/inmunología , Hepatocitos/metabolismo , Factor 1 Regulador del Interferón/metabolismo , Interferón-alfa/metabolismo , Interleucina-6/inmunología , Interleucina-6/metabolismo , Hepatopatías/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Daño por Reperfusión/patología , Factor de Necrosis Tumoral alfa/inmunología , Factor de Necrosis Tumoral alfa/metabolismo , Regulación hacia Arriba/inmunologíaRESUMEN
The role played by prolactin (PRL) in fish immunity is scant. We report here that stimulation of the Atlantic salmon monocytic cell line SHK-1 with native salmon PRL resulted in activation of the respiratory burst and induction of the expression of the genes encoding the phagocyte NADPH oxidase components p47phox, p67phox and gp91phox, and the transcription factor IFN regulatory factor-1 (IRF-1). Interestingly, the pharmacologic inhibition of the Jak/Stat signaling pathway with AG490 blocked reactive oxygen species (ROS) production, and the induction of genes encoding the NADPH oxidase components and IRF-1 in PRL-activated SHK-1 cells. In addition, PRL promoted the phosphorylation of Stat and induced the DNA binding activity of IRF-1. These results, together with the presence of several consensus target motifs for Stat and IRF-1 in the promoter of the tilapia p47phox gene, suggest that PRL regulates p47phox gene expression in fish through the activation of these two key transcription factors. Taken together, our results demonstrate that PRL induces the expression of the genes encoding the major phagocyte NADPH oxidase components and ROS production in fish macrophages via the JAK2/Stat/IRF-1 signaling pathway.
Asunto(s)
Proteínas de Peces/inmunología , Monocitos/inmunología , NADPH Oxidasas/metabolismo , Prolactina/inmunología , Animales , Línea Celular , Regulación de la Expresión Génica/efectos de los fármacos , Factor 1 Regulador del Interferón/genética , Factor 1 Regulador del Interferón/metabolismo , Quinasas Janus/metabolismo , Monocitos/efectos de los fármacos , NADPH Oxidasas/genética , Motivos de Nucleótidos/genética , Especies Reactivas de Oxígeno/metabolismo , Factores de Transcripción STAT/metabolismo , Salmo salar , Transducción de Señal/efectos de los fármacos , Tilapia , Activación Transcripcional , Tirfostinos/farmacologíaRESUMEN
BACKGROUND: Recently, attenuation of anti-inflammatory and increase of pro-inflammatory mediators was demonstrated in individuals with Down syndrome (DS) in comparison with euploid patients during periodontal disease (PD), suggesting a shift to a more aggressive inflammation in DS. AIM: To determine the influence of DS in the modulation of interferons (IFNs) signaling pathway in PD. MATERIALS AND METHODS: Clinical periodontal assessment was performed and gingival tissue samples obtained from a total of 51 subjects, including 19 DS individuals with PD, 20 euploid individuals with PD and 12 euploid individuals without PD. Expression levels of interferon-gamma (IFNG) and interferon-alpha (IFNA), and their receptors IFNGR1, IFNGR2, IFNAR1 and IFNAR2, the signaling intermediates Janus kinase 1 (JAK1), signal transducer and activator of transcription 1 (STAT1) and interferon regulatory factor 1 (IRF1) were determined using real time quantitative polymerase chain reaction (qPCR). RESULTS: Clinical signs of periodontal disease were markedly more severe in DS and euploid patients with PD in comparison to euploid and periodontally healthy patients. There was no difference on mRNA levels of IFNA, IFNG, INFGR2, IFNAR1 and IFNAR2 between DS and euploid individuals, even though some of these genes are located on chromosome 21. STAT1 and IRF1 mRNA levels were significantly lower in DS patients in comparison with euploid individuals with PD. In euploid individuals, PD was associated with an increased expression of IFNGR1, IFNGR2, IFNAR1, STAT1 and IRF1. CONCLUSIONS: Reduced expression of STAT1 and IRF1 genes indicate an impaired activation of IFNs signaling in individuals with DS and PD. Expression of IFNA, IFNG and IFN receptors was not altered in DS patients, indicating that indirect mechanisms are involved in the reduced activation of IFN signaling.
Asunto(s)
Síndrome de Down/genética , Regulación de la Expresión Génica , Interferón-alfa/metabolismo , Interferón gamma/metabolismo , Periodontitis/genética , Adulto , Síndrome de Down/complicaciones , Síndrome de Down/metabolismo , Femenino , Humanos , Factor 1 Regulador del Interferón/metabolismo , Janus Quinasa 1/metabolismo , Masculino , Persona de Mediana Edad , Periodontitis/complicaciones , Periodontitis/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Receptor de Interferón alfa y beta/análisis , Receptor de Interferón alfa y beta/genética , Receptor de Interferón alfa y beta/metabolismo , Receptores de Interferón/genética , Receptores de Interferón/metabolismo , Factor de Transcripción STAT1/metabolismo , Transducción de Señal , Adulto Joven , Receptor de Interferón gammaRESUMEN
Recent investigations postulate a participation of the interferon regulatory factor-1 (IRF-1) in the development of myelodysplasia (MDS) and in the pathogenesis of autoimmune manifestations (AIMs) in patients with this disease. The aim of this prospective study was to compare the IRF-1 immunoexpression in MDS patients with or without AIMs and to investigate its prognostic relevance. Fifty consecutive MDS patients entered this prospective study. There was no difference in overall survival between patients with or without autoimmune manifestations. In a multivariate Cox regression "IRF-1 expression in immature myeloid cells", Hb, and the IPSS risk group stratification were independent prognostic parameters. Bootstrap resampling confirmed these data. In a multivariate logistic regression older patients with, higher platelet count and increased IRF-1 expression had a higher risk to develop autoimmune-like phenomena. Thus our study shows that IRF-1 plays an ambiguous role in MDS patients. Whereas high levels of IRF-1 in myeloid cells are a favorable prognostic factor for overall survival, they increase the probability of the manifestation of autoimmune phenomena, with a diminished quality of life.
Asunto(s)
Enfermedades Autoinmunes/genética , Factor 1 Regulador del Interferón/genética , Síndromes Mielodisplásicos/genética , Anciano , Anciano de 80 o más Años , Femenino , Estudios de Seguimiento , Regulación de la Expresión Génica , Humanos , Factor 1 Regulador del Interferón/metabolismo , Cariotipificación , Masculino , Persona de Mediana Edad , Síndromes Mielodisplásicos/mortalidad , Síndromes Mielodisplásicos/fisiopatología , Síndromes Mielodisplásicos/psicología , Células Mieloides/fisiología , Estudios Prospectivos , Calidad de Vida , Análisis de Regresión , Tasa de Supervivencia , Sobrevivientes , Adulto JovenRESUMEN
El lupus eritematoso sistémico es una enfermedad autoinmune caracterizada por la producción de autoanticuerpos y una diversidad de manifestaciones clínicas. Pese a que su etiología es desconocida, factores genéticos y factores ambientales contribuyen a la pérdida de la tolerancia. Todos los componentes clave del sistema inmune participan en los mecanismos inmunopatogénicos que subyacen a la enfermedad. Esta revisión describe el rol de estos componentes.
Systemic lupus erythematosus is a systemic autoimmune disease characterized by the production of autoantibodies and a diversity of clinical manifestations. Although the etiology of systemic lupus erythematosus is unknown, both genetic and environmental factors contribute to the loss of self-tolerance. All key components of the immune systems are involved in the underlying mechanisms of the disease. This review describes the role of these components.
Asunto(s)
Humanos , Autoanticuerpos/inmunología , Lupus Eritematoso Sistémico/inmunología , Factor 1 Regulador del Interferón/inmunología , Inmunidad Innata , Linfocitos B/inmunología , Linfocitos T/inmunologíaRESUMEN
Deletions on chromosomes 5 and 7 are frequently seen in myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). It is assumed that these deletions indicate loss of tumor suppressor genes on these chromosomes and until these tumor suppressor genes are identified, the functional consequences of these deletions and the molecular basis of these myeloid disorders cannot be completely understood. We evaluated loss of heterozygosity (LOH) in 44 patients (18 MDS and 26 AML, diagnosed according to WHO classification criteria) at diagnosis, using a four-microsatellite marker panel: an intragenic marker on the 7th intron of gene IRF-1 of the 5q31.1 region and three markers located inside the 7q31.1 region and correlated the LOH with karyotype abnormalities. The microsatellites chosen corresponded to chromosome regions frequently deleted in MDS/AML. The samples with Q (peak area) less than or equal to 0.50 were indicative of LOH. The percent of informative samples (i.e., heterozygous) for the intragenic microsatellite in gene IRF-1 and in loci D7S486, D7S515 and D7S522 were 66.6, 73.7, 75.5, and 48.8%, respectively. Cytogenetic abnormalities by G-banding were found in 36% (16/44) of the patients (2 of 18 MDS and 14 of 26 AML patients). We found a significantly positive association of the occurrence of LOH with abnormal karyotype (P < 0.05; chi-square test) and there were cases with LOH but the karyotype was normal (by G-banding). These data indicate that LOH in different microsatellite markers is possibly an event previous to chromosomal abnormalities in these myeloid neoplasias.
Asunto(s)
Aberraciones Cromosómicas , Factor 1 Regulador del Interferón/genética , Leucemia Mieloide Aguda/genética , Pérdida de Heterocigocidad/genética , Síndromes Mielodisplásicos/genética , Marcadores Genéticos , Humanos , Repeticiones de Microsatélite/genética , Reacción en Cadena de la PolimerasaRESUMEN
Deletions on chromosomes 5 and 7 are frequently seen in myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). It is assumed that these deletions indicate loss of tumor suppressor genes on these chromosomes and until these tumor suppressor genes are identified, the functional consequences of these deletions and the molecular basis of these myeloid disorders cannot be completely understood. We evaluated loss of heterozygosity (LOH) in 44 patients (18 MDS and 26 AML, diagnosed according to WHO classification criteria) at diagnosis, using a four-microsatellite marker panel: an intragenic marker on the 7th intron of gene IRF-1 of the 5q31.1 region and three markers located inside the 7q31.1 region and correlated the LOH with karyotype abnormalities. The microsatellites chosen corresponded to chromosome regions frequently deleted in MDS/AML. The samples with Q (peak area) less than or equal to 0.50 were indicative of LOH. The percent of informative samples (i.e., heterozygous) for the intragenic microsatellite in gene IRF-1 and in loci D7S486, D7S515 and D7S522 were 66.6, 73.7, 75.5, and 48.8 percent, respectively. Cytogenetic abnormalities by G-banding were found in 36 percent (16/44) of the patients (2 of 18 MDS and 14 of 26 AML patients). We found a significantly positive association of the occurrence of LOH with abnormal karyotype (P < 0.05; chi-square test) and there were cases with LOH but the karyotype was normal (by G-banding). These data indicate that LOH in different microsatellite markers is possibly an event previous to chromosomal abnormalities in these myeloid neoplasias.
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Humanos , Aberraciones Cromosómicas , Factor 1 Regulador del Interferón/genética , Leucemia Mieloide Aguda/genética , Pérdida de Heterocigocidad/genética , Síndromes Mielodisplásicos/genética , Marcadores Genéticos , Repeticiones de Microsatélite/genética , Reacción en Cadena de la PolimerasaRESUMEN
Brucella species are important zoonotic pathogens affecting a wide variety of mammals. Therefore, the identification of new Brucella virulence factors is of great interest in understanding bacterial pathogenesis and immune evasion. In this study, we have identified Brucella abortus vacB gene that presents 2343 nucleotides and 781 amino acids and it shows 39% identity with Shigella flexneri vacB gene that encodes an exoribonuclease RNase R involved in bacterial virulence. Further, we have inactivated Brucella vacB by gene replacement strategy generating a deletion mutant strain. In order to test the role of Brucella vacB in pathogenesis, BALB/c and interferon regulatory factor-1 (IRF-1) knockout (KO) mice received Brucella vacB mutant, the virulent parental strain 2308 or the vaccine strain RB51 and the bacterial CFU numbers in spleens and mous survival were monitored. Our results demonstrated that the B. abortus DeltavacB mutant and the wild type strain 2308 showed similar CFU numbers in BALB/c mice. Additionally, IRF-1 KO mice that received either the vacB mutant or S2308 strain died in 12-14 days postinfection; in contrast, all animals that received the RB51 vaccine strain survived for 30 days postinoculation. In summary, this study reports that the vacB gene in B. abortus has no impact on bacterial pathogenesis.
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Proteínas Bacterianas/genética , Brucella abortus/patogenicidad , Exorribonucleasas/genética , Secuencia de Aminoácidos , Animales , Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Secuencia de Bases , Vacuna contra la Brucelosis/administración & dosificación , Brucella abortus/genética , Brucella abortus/metabolismo , Brucelosis/inmunología , Brucelosis/microbiología , Brucelosis/mortalidad , Brucelosis/prevención & control , Exorribonucleasas/química , Exorribonucleasas/metabolismo , Femenino , Eliminación de Gen , Humanos , Factor 1 Regulador del Interferón/genética , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Datos de Secuencia Molecular , Análisis de Secuencia de ADN , Bazo/microbiología , VirulenciaRESUMEN
Using DDRT-PCR, we compared the mRNA content of untreated and TNF-treated mouse embryonic fibroblasts (MEFs). Among differentially represented fragments, we identified and cloned a novel TNF-stimulated gene named Tsg-5. This gene, mapped to mouse chromosome 14, has three exons that can be alternatively spliced giving rise to two mRNA species, one spanning three exons and another that skips the second exon. Analysis of full-length Tsg-5 cDNA revealed a potential start codon within exon 2 encoding an ORF of 40 amino-acids. No homology with known mouse or human sequences, neither at the nucleotide nor at the amino-acid level could be found in public databases. In MEFs, Tsg-5 is induced by tumor necrosis factor-alpha (TNF) and IL-1 beta, albeit with distinct kinetics. TNF-induced Tsg-5 expression is NF-kappa B-dependent as it was inhibited by MG132, lactacystin, Bay 11-7083, and Bay 11-7085. Analysis of Tsg-5 expression in vivo revealed that the gene and its encoded polypeptide are constitutively expressed in the thymus and ovary, whereas, in LPS-treated mice, Tsg-5 mRNA can be detected in the spleen, lung, and brain. Our data suggest that Tsg-5 encodes a new, rare transcript, with a very tight regulation of expression and differential splicing.
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Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/aislamiento & purificación , Factor de Necrosis Tumoral alfa/fisiología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Células Cultivadas , Clonación Molecular , ADN Complementario/aislamiento & purificación , Proteínas de Unión al ADN/antagonistas & inhibidores , Proteínas de Unión al ADN/biosíntesis , Proteínas de Unión al ADN/deficiencia , Proteínas de Unión al ADN/fisiología , Femenino , Regulación de la Expresión Génica/inmunología , Factor 1 Regulador del Interferón , Ratones , Ratones Endogámicos BALB C , Ratones Noqueados , Datos de Secuencia Molecular , FN-kappa B/metabolismo , Especificidad de Órganos/genética , Especificidad de Órganos/inmunología , Fosfoproteínas/biosíntesis , Fosfoproteínas/deficiencia , Fosfoproteínas/genética , Reacción en Cadena de la PolimerasaRESUMEN
The induction of indoleamine 2,3-dioxygenase (INDO) expression and the tryptophan (Trp)-kynurenine (Kyn) metabolic pathway during in vivo infection with Toxoplasma gondii was investigated. Decreased levels of Trp and increased formation of Kyn were observed in the lungs, brain, and serum from mice infected with T. gondii. Maximal INDO mRNA expression and enzyme activity were detected in the lungs at 10 to 20 days postinfection. Further, the induction of INDO mRNA expression, Trp degradation and Kyn formation were completely absent in tissues from mice deficient in IFN-gamma (IFN-gamma(-/-)) or IFN regulatory factor -1 (IRF-1(-/-)). These findings indicate the important role of endogenous IFN-gamma and IRF-1 in the in vivo induction of the Trp-Kyn metabolic pathway during acute infection with T. gondii. In contrast, expression of INDO mRNA and its activity was preserved in the tissues of TNF-receptor p55- or inducible nitric oxide synthase-deficient mice infected with T. gondii. Together with the results showing the extreme susceptibility of the IFN-gamma(-/-) and the IRF-1(-/-) mice to infection with T. gondii, our results indicate a possible involvement of INDO and Trp degradation in host resistance to early infection with this parasite.
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Proteínas de Unión al ADN/fisiología , Expresión Génica , Interferón gamma/fisiología , Quinurenina/biosíntesis , Fosfoproteínas/fisiología , Toxoplasmosis/metabolismo , Triptófano Oxigenasa/genética , Triptófano/metabolismo , Enfermedad Aguda , Animales , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/inmunología , Susceptibilidad a Enfermedades , Factor 1 Regulador del Interferón , Interferón gamma/genética , Interferón gamma/inmunología , Cinética , Quinurenina/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Fosfoproteínas/genética , Fosfoproteínas/inmunología , ARN Mensajero , Toxoplasma/inmunología , Toxoplasmosis/enzimología , Toxoplasmosis/inmunología , Triptófano/inmunología , Triptófano Oxigenasa/inmunología , Triptófano Oxigenasa/metabolismoRESUMEN
Prolactin (PRL) is a versatile hormone that is produced by the anterior pituitary gland and various extrapituitary sites including immune cells. Furthermore, PRL has widespread influences on proliferation and differentiation of a variety of cells in the immune system and is, in effect, a cytokine. PRL-receptors (PRL-R) are distributed throughout the immune system and are included as members of the cytokine receptor superfamily. PRL-R signal transduction is mediated by a complex array of signaling molecules of which JAK2, Stat1 and Stat5 pathway have been well studied. In PRL-stimulated T cells, the transcription factor gene, interferon regulatory factor-1 provides a mechanism whereby PRL can regulate the immune response. The human PRL gene is situated on the short arm of chromosome 6 close to the major histocompatibility complex. Polymorphisms of the human PRL gene have implications for production of lymphocyte PRL in SLE. Mild and moderate hyperprolactinemia (HPRL) has been demonstrated in 20-30% of SLE patients and is associated with active disease. HPRL may have a role in lupus nephritis and central nervous system involvement of SLE patients. HPRL stimulated the production of autoantibodies. These evidences support the important role of PRL in autoimmunity and autoimmune diseases, mainly SLE.
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Autoinmunidad , Proteínas de la Leche , Prolactina/inmunología , Proteínas Proto-Oncogénicas , Animales , Proteínas de Unión al ADN/fisiología , Humanos , Hiperprolactinemia/complicaciones , Hiperprolactinemia/inmunología , Inmunogenética , Factor 1 Regulador del Interferón , Janus Quinasa 2 , Lupus Eritematoso Sistémico/complicaciones , Lupus Eritematoso Sistémico/etiología , Lupus Eritematoso Sistémico/inmunología , Fosfoproteínas/fisiología , Prolactina/genética , Prolactina/fisiología , Proteínas Tirosina Quinasas/fisiología , Receptores de Prolactina/fisiología , Factor de Transcripción STAT5 , Transducción de Señal , Transactivadores/fisiologíaRESUMEN
Syngeneic IFN-gamma(-/-) and IRF-1(-/-) mice are very sensitive to B16F10-Nex2 murine melanoma cells implanted subcutaneously. In contrast, IFN-gamma-R(-/-) (GRKO) mice are remarkably resistant to tumor development. Only 0-30% of these animals, challenged with a high dose of melanoma cells (5 x 10(5)), developed tumors at a late stage. The hypothesis of interferon gamma (IFN-gamma) accumulation and consequent cytotoxicity to implanted tumor cells was confirmed in vitro and ex vivo. IFN-gamma reduced tumor-cell growth in vitro in 60-81%, added alone or with LPS. Splenocytes and peritoneal macrophages from naïve GRKO mice activated with anti-CD3 and interleukin-12 (IL-12), respectively, accumulated IFN-gamma at levels 10-fold those of the wild-type. Supernatants of IL-12-activated macrophages from GRKO mice were toxic to B16F10-Nex2 cells, an effect reversible by anti-IFN-gamma antibody treatment. IL-12-activated macrophages from iNOS(-/-) mice were still highly cytotoxic to B16F10-Nex2 cells, but IL-12-activated macrophages from IFN-gamma-deficient mice were not inhibitory. In vivo, a single injection of anti-IFN-gamma antibody 18 h after tumor-cell challenge in GRKO mice rendered all animals susceptible to B16F10-Nex2 melanoma development. No tumors developed in the untreated GRKO mice during up to 45 days of observation. This model can be useful in understanding immune responses that involve IFN-gamma as a direct cytotoxic factor.