RESUMEN
Although numerous environmental exposures have been suggested as triggers for preclinical autoimmunity, only a few have been confidently linked to autoimmune diseases. For disease-associated exposures, the lung is a common site where chronic exposure results in cellular toxicity, tissue damage, inflammation, and fibrosis. These features are exacerbated by exposures to particulate material, which hampers clearance and degradation, thus facilitating persistent inflammation. Coincident with exposure and resulting pathological processes is the posttranslational modification of self-antigens, which, in concert with the formation of tertiary lymphoid structures containing abundant B cells, is thought to promote the generation of autoantibodies that in some instances demonstrate major histocompatibility complex restriction. Under appropriate gene-environment interactions, these responses can have diagnostic specificity. Greater insight into the molecular and cellular requirements governing this process, especially those that distinguish preclinical autoimmunity from clinical autoimmunedisease, may facilitate determination of the significance of environmental exposures in human autoimmune disease.
Asunto(s)
Enfermedades Autoinmunes , Autoinmunidad , Autoanticuerpos , Exposición a Riesgos Ambientales , Humanos , InflamaciónRESUMEN
Exposure to heavy metals may have toxic effects on several human organs causing morbidity and mortality. Metals may trigger or exacerbate autoimmunity in humans. Inbred mouse strains with certain H-2 haplotypes are susceptible to xenobiotic-induced autoimmunity; and their immune response to metals such as mercury, gold, and silver have been explored. Serum antinuclear antibodies (ANA), polyclonal B-cell activation, hypergammaglobulinemia and tissue immune complex deposition are the main features of metal-induced autoimmunity in inbred mice. However, inbred mouse strains do not represent the genetic heterogeneity in humans. In this study, outbred Swiss Webster (SW) mice exposed to gold or mercury salts showed immune and autoimmune responses. Intramuscular injection of 22.5 mg/kg.bw aurothiomalate (AuTM) induced IgG ANA in SW mice starting after 5 weeks that persisted until week 15 although with a lower intensity. This was accompanied by elevated serum levels of total IgG antibodies against chromatin and total histones. Exposure to gold led to development of serum IgG autoantibodies corresponding to H1 and H2A histones, and dsDNA. Both gold and mercury induced polyclonal B-cell activation. Eight mg/L mercuric chloride (HgCl2) in drinking water, caused IgG antinucleolar antibodies (ANoA) after 5 weeks in SW mice accompanied by immune complex deposition in kidneys and spleen. Serum IgG antibodies corresponding to anti-fibrillarin, and anti-PM/Scl-100 antibodies, were observed in mercury-exposed SW mice. Gold and mercury trigger systemic autoimmune response in genetically heterogeneous outbred SW mice and suggest them as an appropriate model to study xenobiotic-induced autoimmunity.
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Anticuerpos Antinucleares/sangre , Autoinmunidad/efectos de los fármacos , Linfocitos B/efectos de los fármacos , Tiomalato Sódico de Oro/toxicidad , Inmunoglobulina G/sangre , Activación de Linfocitos/efectos de los fármacos , Cloruro de Mercurio/toxicidad , Administración Oral , Animales , Complejo Antígeno-Anticuerpo , Linfocitos B/inmunología , Linfocitos B/metabolismo , Cromatina/inmunología , Proteínas Cromosómicas no Histona/inmunología , Exorribonucleasas/inmunología , Complejo Multienzimático de Ribonucleasas del Exosoma/inmunología , Femenino , Tiomalato Sódico de Oro/administración & dosificación , Histonas/inmunología , Inyecciones Intramusculares , Riñón/efectos de los fármacos , Riñón/inmunología , Cloruro de Mercurio/administración & dosificación , Ratones , Bazo/efectos de los fármacos , Bazo/inmunologíaRESUMEN
Type I IFN and nucleic acid-sensing TLRs are both strongly implicated in the pathogenesis of lupus, with most patients expressing IFN-induced genes in peripheral blood cells and with TLRs promoting type I IFNs and autoreactive B cells. About a third of systemic lupus erythematosus patients, however, lack the IFN signature, suggesting the possibility of type I IFN-independent mechanisms. In this study, we examined the role of type I IFN and TLR trafficking and signaling in xenobiotic systemic mercury-induced autoimmunity (HgIA). Strikingly, autoantibody production in HgIA was not dependent on the type I IFN receptor even in NZB mice that require type I IFN signaling for spontaneous disease, but was dependent on the endosomal TLR transporter UNC93B1 and the endosomal proton transporter, solute carrier family 15, member 4. HgIA also required the adaptor protein-3 complex, which transports TLRs from the early endosome to the late endolysosomal compartments. Examination of TLR signaling pathways implicated the canonical NF-κB pathway and the proinflammatory cytokine IL-6 in autoantibody production, but not IFN regulatory factor 7. These findings identify HgIA as a novel type I IFN-independent model of systemic autoimmunity and implicate TLR-mediated NF-κB proinflammatory signaling from the late endocytic pathway compartments in autoantibody generation.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Endosomas/metabolismo , Lupus Eritematoso Sistémico/inmunología , Proteínas de Transporte de Membrana/metabolismo , Receptores Toll-Like/metabolismo , Animales , Autoanticuerpos/metabolismo , Enfermedades Autoinmunes/inducido químicamente , Autoinmunidad , Células Cultivadas , Femenino , Humanos , Interferón Tipo I/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Lisosomas/metabolismo , Masculino , Proteínas de Transporte de Membrana/genética , Mercurio , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos NZB , Ratones Noqueados , FN-kappa B/genética , FN-kappa B/metabolismo , Transporte de Proteínas , Receptor de Interferón alfa y beta/genética , Transducción de Señal , Receptores Toll-Like/genética , XenobióticosRESUMEN
BACKGROUND: Urinary bladder carcinoma stage T1 is an unpredictable disease that in some cases has a good prognosis with only local or no recurrence, but in others can appear as a more aggressive tumor with progression to more advanced stages. The aim here was to investigate stage T1 tumors regarding MDM2 promoter SNP309 polymorphism, mutations in the p53 gene, and expression of p53 and p16 measured by immunohistochemistry, and subsequently relate these changes to tumor recurrence and progression. We examined a cohort of patients with primary stage T1 urothelial carcinoma of the bladder and their tumors. METHODS: After re-evaluation of the original slides and exclusions, the study population comprised 141 patients, all with primary stage T1 urothelial carcinoma of the bladder. The hospital records were screened for clinical parameters and information concerning presence of histologically proven recurrence and progression. The paraffin-embedded tumor material was evaluated by immunohistochemistry. Any mutations found in the p53 gene were studied by single-strand conformation analysis and Sanger sequencing. The MDM2 SNP309 polymorphism was investigated by pyrosequencing. Multivariate analyses concerning association with prognosis were performed, and Kaplan-Meier analysis was conducted for a combination of changes and time to progression. RESULTS: Of the 141 patients, 82 had at least one MDM2 SNP309 G allele, and 53 had a mutation in the p53 gene, but neither of those anomalies was associated with a worse prognosis. A mutation in the p53 gene was associated with immunohistochemically visualized p53 protein expression at a cut-off value of 50%. In the group with p53 mutation Kaplan-Meier analysis showed higher rate of progression and shorter time to progression in patients with immunohistochemically abnormal p16 expression compared to them with normal p16 expression (p = 0.038). CONCLUSIONS: MDM2 SNP309 promoter polymorphism and mutations in p53 were not associated with worse prognosis in this cohort of patients with primary stage T1 urinary bladder carcinoma. However, patients with abnormal p16 expression and a mutated p53 gene had a higher rate of and a shorter time to progression, and p53 gene mutation was associated with an abnormal immunohistochemistry for p53 at a cut-off of 50%.
Asunto(s)
Carcinoma de Células Transicionales/genética , Genes p53/genética , Proteínas Proto-Oncogénicas c-mdm2/genética , Proteína p53 Supresora de Tumor/metabolismo , Neoplasias de la Vejiga Urinaria/genética , Adulto , Anciano , Anciano de 80 o más Años , Carcinoma de Células Transicionales/patología , Estudios de Cohortes , Inhibidor p16 de la Quinasa Dependiente de Ciclina , Inhibidor p21 de las Quinasas Dependientes de la Ciclina/metabolismo , Femenino , Genotipo , Humanos , Inmunohistoquímica , Estimación de Kaplan-Meier , Masculino , Persona de Mediana Edad , Proteínas de Neoplasias/metabolismo , Estadificación de Neoplasias , Reacción en Cadena de la Polimerasa , Polimorfismo de Nucleótido Simple , Pronóstico , Regiones Promotoras Genéticas , Modelos de Riesgos Proporcionales , Proteína de Retinoblastoma/metabolismo , Neoplasias de la Vejiga Urinaria/patologíaRESUMEN
Exposure to mercury (Hg) and silver (Ag) has been shown to induce autoimmune diseases in genetically susceptible rodents. Here, A.SW mice were initially exposed to HgCl2, AgNO3 or tap water (control) for 3 weeks. After 13 weeks of stoppage, all mice had secondary exposure to 203HgCl2. After secondary exposure, higher and earlier ANoA titers were observed in mice initially exposed to Hg or Ag compared to control. Further, mice initially exposed to Ag showed higher total IgG1 and IgG2a, Whole Body Retention and lymph nodes and spleen accumulation of Hg compared to mice initially exposed to Hg and controls. These findings showed an earlier and stronger immunological response in A.SW mice compared with control, following re-exposure to heavy metals indicating an immunological memory. Additionally, secondary exposure to a different heavy metal may aggravate the effects of exposure of at least one of the metals indicating cross-reactivity.
Asunto(s)
Enfermedades Autoinmunes , Mercurio , Ratones , Animales , Autoinmunidad , Enfermedades Autoinmunes/inducido químicamente , Enfermedades Autoinmunes/genética , Predisposición Genética a la Enfermedad , Inmunoglobulina G/farmacología , AceleraciónRESUMEN
IFN-γ is essential for idiopathic and murine mercury-induced systemic autoimmunity (mHgIA), and heterozygous IFN-γ(+/-) mice also exhibit reduced disease. This suggests that blocking specific IFN-γ-related pathways that may only partially inhibit IFN-γ production or function will also suppress autoimmunity. To test this hypothesis, mice deficient in genes regulating IFN-γ expression (Casp1, Nlrp3, Il12a, Il12b, Stat4) or function (Ifngr1, Irf1) were examined for mHgIA susceptibility. Absence of either Ifngr1 or Irf1 resulted in a striking reduction of disease, while deficiency of genes promoting IFN-γ expression had modest to no effect. Furthermore, both Irf1- and Ifng-deficiency only modestly reduced the expansion of CD44(hi) and CD44(hi)CD55(lo) CD4(+) T cells, indicating that they are not absolutely required for T cell activation. Thus, there is substantial redundancy in genes that regulate IFN-γ expression in contrast to those that mediate later signaling events. These findings have implications for the therapeutic targeting of IFN-γ pathways in systemic autoimmunity.
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Enfermedades Autoinmunes/genética , Autoinmunidad/genética , Regulación de la Expresión Génica/inmunología , Interferón gamma/genética , Transducción de Señal/inmunología , Linfocitos T/inmunología , Animales , Antígenos CD/genética , Antígenos CD/inmunología , Enfermedades Autoinmunes/inducido químicamente , Enfermedades Autoinmunes/metabolismo , Enfermedades Autoinmunes/patología , Proliferación Celular , Eliminación de Gen , Heterocigoto , Inyecciones Subcutáneas , Interferón gamma/inmunología , Interferón gamma/metabolismo , Cloruro de Mercurio , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Transducción de Señal/genética , Linfocitos T/patologíaRESUMEN
Epidermolysis bullosa acquisita is a prototypical organ-specific autoimmune disease caused by autoantibodies against type VII collagen of the dermal-epidermal junction. Although mechanisms of autoantibody-induced blister formation were extensively characterized, the initiation of autoantibody production in autoimmune blistering diseases is still poorly defined. In the current study, we addressed the role of T cells for the production of blister-inducing autoantibodies in mice immunized with type VII collagen. To detect autoreactive type VII collagen-specific T cells, lymph node cells from immunized SJL mice were stimulated in vitro with recombinant Ag, and their proliferation was measured by radioactive thymidine incorporation and flow cytometry analysis of CFSE-labeled cells. Interestingly, using synthetic peptides of the immunogen, partly different T and B cell epitopes in mice immunized with type VII collagen were demonstrated. In contrast to wild-type mice, immunization with type VII collagen of SJL athymic nude mice lacking T cells did not induce an autoimmune response and blistering phenotype. Importantly, SJL nude mice repleted with T cells from immunized wild-type mice showed a robust and durable autoantibody production resulting in subepidermal blistering disease in the recipients. Our present results demonstrate that T cells are required for the initiation of autoimmunity against type VII collagen in experimental epidermolysis bullosa acquisita and provide a basis for developing T cell-directed immunomodulatory strategies for this and related autoimmune diseases.
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Autoanticuerpos/biosíntesis , Vesícula/inmunología , Colágeno Tipo VII/inmunología , Epidermólisis Ampollosa Adquirida/inmunología , Subgrupos de Linfocitos T/inmunología , Secuencia de Aminoácidos , Animales , Autoanticuerpos/efectos adversos , Vesícula/patología , Células Cultivadas , Colágeno Tipo VII/administración & dosificación , Modelos Animales de Enfermedad , Epidermólisis Ampollosa Adquirida/patología , Epítopos de Linfocito T/inmunología , Epítopos de Linfocito T/metabolismo , Activación de Linfocitos/inmunología , Ratones , Ratones Endogámicos , Ratones Desnudos , Datos de Secuencia Molecular , Subgrupos de Linfocitos T/metabolismo , Subgrupos de Linfocitos T/patologíaRESUMEN
OBJECTIVE: The HER2 receptor is involved in pathways essential for cell proliferation, and is an important predictive and prognostic factor in breast cancer. HER2 probably plays a critical role in many types of cancer, including urothelial carcinoma of the bladder (UCB). Stage T1 UCB exhibits heterogeneous clinical behaviour, and the frequency of HER2 expression in such disease has not been thoroughly examined. The aim of this study was to use an immunohistochemical technique to evaluate the frequency of HER2 expression in a defined population-based cohort of patients registered as having primary stage T1 UCB. MATERIAL AND METHODS: The initial study population comprised 285 patients registered as having primary stage T1 UCB. The original histological specimens were re-evaluated with regard to T stage and World Health Organization grade. Hospital records provided information on tumour size, multiplicity, possible presence of histologically proven recurrence and progression. The patients were followed for at least 5 years or until death. In tumours still considered stage T1 after re-evaluation, HER2 was investigated by immunohistochemistry of paraffin-embedded material and scored according to the guidelines used in breast cancer. RESULTS: After histopathological re-evaluation, 201 patients were still T1 UCB and could be investigated regarding HER2 expression. HER2 overexpression was observed in 25 of those patients (12.4%). HER2 status was not significantly associated with recurrence or progression. CONCLUSIONS: HER2 was overexpressed in 12.4% of the present cohort of patients with primary stage T1 UCB. There was no significant association between tumour HER2 status and prognosis.
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Carcinoma/metabolismo , Carcinoma/patología , Progresión de la Enfermedad , Recurrencia Local de Neoplasia/metabolismo , Receptor ErbB-2/metabolismo , Neoplasias de la Vejiga Urinaria/metabolismo , Neoplasias de la Vejiga Urinaria/patología , Anciano , Femenino , Humanos , Modelos Logísticos , Estudios Longitudinales , Masculino , Estadificación de Neoplasias , Carga TumoralRESUMEN
Mercury (Hg) exposure from dental amalgam fillings and thimerosal in vaccines is not a major health hazard, but adverse health effects cannot be ruled out in a small and more susceptible part of the exposed population. Individual differences in toxicokinetics may explain susceptibility to mercury. Inbred, H-2-congenic A.SW and B10.S mice and their F1- and F2-hybrids were given HgCl2 with 2.0 mg Hg/L drinking water and traces of (203)Hg. Whole-body retention (WBR) was monitored until steady state after 5 weeks, when the organ Hg content was assessed. Despite similar Hg intake, A.SW males attained a 20-30% significantly higher WBR and 2- to 5-fold higher total renal Hg retention/concentration than A.SW females and B10.S mice. A selective renal Hg accumulation but of lower magnitude was seen also in B10.S males compared with females. Differences in WBR and organ Hg accumulation are therefore regulated by non-H-2 genes and gender. Lymph nodes lacked the strain- and gender-dependent Hg accumulation profile of kidney, liver and spleen. After 15 days without Hg A.SW mice showed a 4-fold higher WBR and liver Hg concentration, but 11-fold higher renal Hg concentration, showing the key role for the kidneys in explaining the slower Hg elimination in A.SW mice. The trait causing higher mercury accumulation was not dominantly inherited in the F1 hybrids. F2 mice showed a large inter-individual variation in Hg accumulation, showing that multiple genetic factors influence the Hg toxicokinetics in the mouse. The genetically heterogeneous human population may therefore show a large variation in mercury toxicokinetics.
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Cloruro de Mercurio/farmacocinética , Cloruro de Mercurio/toxicidad , Animales , Femenino , Expresión Génica/efectos de los fármacos , Glutatión/metabolismo , Glutatión Transferasa/metabolismo , Semivida , Masculino , Metalotioneína/biosíntesis , Metalotioneína/genética , Ratones , Caracteres Sexuales , Especificidad de la Especie , Distribución Tisular , Abastecimiento de Agua/análisisRESUMEN
Susceptibility to most autoimmune diseases is dependent on polygenic inheritance, environmental factors, and poorly defined stochastic events. One of the significant challenges facing autoimmune disease research is in identifying the specific events that trigger loss of tolerance and autoimmunity. Although many intrinsic factors, including age, sex, and genetics, contribute to autoimmunity, extrinsic factors such as drugs, chemicals, microbes, or other environmental factors can also act as important initiators. This review explores how certain extrinsic factors, namely, drugs and chemicals, can promote the development of autoimmunity, focusing on a few better characterized agents that, in most instances, have been shown to produce autoimmune manifestations in human populations. Mechanisms of autoimmune disease induction are discussed in terms of research obtained using specific animal models. Although a number of different pathways have been delineated for drug/chemical-induced autoimmunity, some similarities do exist, and a working model is proposed.
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Enfermedades Autoinmunes/inducido químicamente , Autoinmunidad , Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos , Animales , Autoinmunidad/efectos de los fármacos , HumanosRESUMEN
INTRODUCTION: AA amyloidosis develops as a result of prolonged inflammation and is characterized by deposits of N-terminal proteolytic fragments of the acute phase reactant serum amyloid A (SAA). Macrophages are usually found adjacent to amyloid, suggesting their involvement in the formation and/or degradation of the amyloid fibrils. Furthermore, accumulating evidence suggests that lipid membranes accelerate the fibrillation of different amyloid proteins. METHODS: Using an experimental mouse model of AA amyloidosis, we compared the amyloidogenic effect of liposomes and/or amyloid-enhancing factor (AEF). Inflammation was induced by subcutaneous injection of silver nitrate followed by intravenous injection of liposomes and/or AEF to accelerate amyloid formation. RESULTS: We showed that liposomes accelerate amyloid formation in inflamed mice, but the amyloidogenic effect of liposomes was weaker compared with AEF. Regardless of the induction method, amyloid deposits were mainly found in the marginal zones of the spleen and coincided with the depletion of marginal zone macrophages, while red pulp macrophages and metallophilic marginal zone macrophages proved insensitive to amyloid deposition. CONCLUSIONS: We conclude that increased intracellular lipid content facilitates AA amyloid fibril formation and show that the mouse model of AA amyloidosis is a suitable system for further mechanistic studies.
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Amiloidosis/metabolismo , Inflamación , Lípidos , Macrófagos/metabolismo , Proteína Amiloide A Sérica/metabolismo , Amiloidosis/patología , Animales , Modelos Animales de Enfermedad , Femenino , Macrófagos/inmunología , Ratones , Agregación Patológica de ProteínasRESUMEN
BACKGROUND: Human exposure to mercury leads to a variety of pathologies involving numerous organ systems including the immune system. A paucity of epidemiological studies and suitable diagnostic criteria, however, has hampered collection of sufficient data to support a causative role for mercury in autoimmune diseases. Nevertheless, there is evidence that mercury exposure in humans is linked to markers of inflammation and autoimmunity. This is supported by experimental animal model studies, which convincingly demonstrate the biological plausibility of mercury as a factor in the pathogenesis of autoimmune disease. SCOPE OF THE REVIEW: In this review, we focus on ability of mercury to elicit inflammatory and autoimmune responses in both humans and experimental animal models. MAJOR CONCLUSIONS: Although subtle differences exist, the inflammatory and autoimmune responses elicited by mercury exposure in humans and experimental animal models show many similarities. Proinflammatory cytokine expression, lymphoproliferation, autoantibody production, and nephropathy are common outcomes. Animal studies have revealed significant strain dependent differences in inflammation and autoimmunity suggesting genetic regulation. This has been confirmed by the requirement for individual genes as well as genome wide association studies. Importantly, many of the genes required for mercury-induced inflammation and autoimmunity are also required for idiopathic systemic autoimmunity. A notable difference is that mercury-induced autoimmunity does not require type I IFN. This observation suggests that mercury-induced autoimmunity may arise by both common and specific pathways, thereby raising the possibility of devising criteria for environmentally associated autoimmunity. GENERAL SIGNIFICANCE: Mercury exposure likely contributes to the pathogenesis of autoimmunity.
Asunto(s)
Enfermedades Autoinmunes , Autoinmunidad/efectos de los fármacos , Regulación de la Expresión Génica , Mercurio/toxicidad , Animales , Enfermedades Autoinmunes/inducido químicamente , Enfermedades Autoinmunes/genética , Enfermedades Autoinmunes/patología , Citocinas/genética , Citocinas/inmunología , Modelos Animales de Enfermedad , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/inmunología , Estudio de Asociación del Genoma Completo , Humanos , Inflamación/inducido químicamente , Inflamación/inmunología , Inflamación/patologíaRESUMEN
Systemic autoimmune rheumatic disorders (SARD) represent important causes of morbidity and mortality in humans. The mechanisms triggering autoimmune responses are complex and involve a network of genetic factors. Mercury-induced autoimmunity (HgIA) in mice is an established model to study the mechanisms of the development of antinuclear antibodies (ANA), which is a hallmark in the diagnosis of SARD. A.SW mice with HgIA show a significantly higher titer of antinucleolar antibodies (ANoA) than the B10.S mice, although both share the same MHC class II (H-2). We applied a genome-wide association study (GWAS) to their Hg-exposed F2 offspring to investigate the non-MHC genes involved in the development of ANoA. Quantitative trait locus (QTL) analysis showed a peak logarithm of odds ratio (LOD) score of 3.05 on chromosome 3. Microsatellites were used for haplotyping, and fine mapping was conducted with next generation sequencing. The candidate genes Bank1 (B-cell scaffold protein with ankyrin repeats 1) and Nfkb1 (nuclear factor kappa B subunit 1) were identified by additional QTL analysis. Expression of the Bank1 and Nfkb1 genes and their downstream target genes involved in the intracellular pathway (Tlr9, Il6, Tnf) was investigated in mercury-exposed A.SW and B10.S mice by real-time PCR. Bank1 showed significantly lower gene expression in the A.SW strain after Hg-exposure, whereas the B10.S strain showed no significant difference. Nfkb1, Tlr9, Il6 and Tnf had significantly higher gene expression in the A.SW strain after Hg-exposure, while the B10.S strain showed no difference. This study supports the roles of Bank1 (produced mainly in B-cells) and Nfkb1 (produced in most immune cells) as key regulators of ANoA development in HgIA.
Asunto(s)
Proteínas Adaptadoras Transductoras de Señales/metabolismo , Anticuerpos Antinucleares/inmunología , FN-kappa B/metabolismo , Proteínas Adaptadoras Transductoras de Señales/química , Proteínas Adaptadoras Transductoras de Señales/genética , Animales , Linfocitos B/citología , Linfocitos B/inmunología , Linfocitos B/metabolismo , Femenino , Regulación de la Expresión Génica/genética , Haplotipos , Espacio Intracelular/metabolismo , Ratones , Mutación Missense , Polimorfismo de Nucleótido Simple , Receptor Toll-Like 9/metabolismoRESUMEN
Inorganic mercury (mercuric chloride--HgCl(2)) induces in mice an autoimmune syndrome (HgIA) with T cell-dependent polyclonal B cell activation and hypergammaglobulinemia, dose- and H-2-dependent production of autoantibodies targeting the 34 kDa nucleolar protein fibrillarin (AFA), and systemic immune-complex deposits. The organic mercury species methylmercury (MeHg) and ethylmercury (EtHg--in the form of thimerosal) induce AFA, while the other manifestations of HgIA seen after treatment with HgCl(2) are present to varying extent. Since these organic Hg species are converted to the autoimmunogen Hg(2+) in the body, their primary autoimmunogen potential is uncertain and the subject of this study. A moderate dose of HgCl(2) (8 mg/L drinking water--internal dose 148 micro gHg/kg body weight [bw]/day) caused the fastest AFA response, while the induction was delayed after higher (25 mg/L) and lower (1.5 and 3 mg/L) doses. The lowest dose of HgCl(2) inducing AFA was 1.5 mg/L drinking water which corresponded to a renal Hg(2+) concentration of 0.53 micro g/g. Using a dose of 8 mg HgCl(2)/L this threshold concentration was reached within 24 h, and a consistent AFA response developed after 8-10 days. The time lag for the immunological part of the reaction leading to a consistent AFA response was therefore 7-9 days. A dose of thimerosal close to the threshold dose for induction of AFA (2 mg/L drinking water--internal dose 118 micro gHg/kg bw per day), caused a renal Hg(2+) concentration of 1.8 micro g/g. The autoimmunogen effect of EtHg might therefore be entirely due to Hg(2+) formed from EtHg in the body. The effect of organic and inorganic Hg species on T-helper type 1 and type 2 cells during induction of AFA was assessed as the presence and titre of AFA of the IgG1 and IgG2a isotype, respectively. EtHg induced a persistent Th1-skewed response irrespectively of the dose and time used. A low daily dose of HgCl(2) (1.5-3 mg/L) caused a Th1-skewed AFA response, while a moderate dose (8 mg/L) after 2 weeks resulted in a balanced or even Th2-skewed response. Higher daily doses of HgCl(2) (25 mg/L) caused a balanced Th2-Th1 response already from onset. In conclusion, while metabolically formed Hg(2+) might be the main AFA-inducing factor also after treatment with EtHg, the quality of the Hg-induced AFA response is modified by the species of Hg as well as the dose.
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Autoinmunidad/inmunología , Activación de Linfocitos/efectos de los fármacos , Cloruro de Mercurio/toxicidad , Linfocitos T Colaboradores-Inductores/efectos de los fármacos , Animales , Anticuerpos Antinucleares/inmunología , Proteínas Cromosómicas no Histona/sangre , Relación Dosis-Respuesta a Droga , Femenino , Técnica del Anticuerpo Fluorescente , Inmunoglobulina G/clasificación , Inmunoglobulina G/inmunología , Inmunoglobulina G/metabolismo , Riñón/química , Riñón/efectos de los fármacos , Activación de Linfocitos/inmunología , Cloruro de Mercurio/química , Cloruro de Mercurio/inmunología , Ratones , Proteínas Nucleares/sangre , Órbita/irrigación sanguínea , Linfocitos T Colaboradores-Inductores/inmunología , Células TH1/efectos de los fármacos , Células TH1/inmunología , Células Th2/efectos de los fármacos , Células Th2/inmunología , Timerosal/química , Timerosal/inmunología , Timerosal/toxicidad , Factores de TiempoRESUMEN
The oral thrombin inhibitor ximelagatran was withdrawn in the late clinical trial phase because it adversely affected the liver. In approximately 8% of treated patients, drug-induced liver injury (DILI) was expressed as transient alanine transaminase (ALT) elevations. No evidence of DILI had been revealed in the pre-clinical in vivo studies. A whole genome scan study performed on the clinical study material identified a strong genetic association between the major histocompatibility complex alleles for human leucocyte antigens (HLA) (HLA-DR7 and HLA-DQ2) and elevated ALT levels in treated patients. An immune-mediated pathogenesis was suggested. Here, we evaluated whether HLA transgenic mice models could be used to investigate whether the expression of relevant HLA molecules was enough to reproduce the DILI effects in humans. In silico modelling performed in this study revealed association of both ximelagatran (pro-drug) and melagatran (active drug) to the antigen-presenting groove of the homology modelled HLA-DR7 molecule suggesting "altered repertoire" as a key initiating event driving development of DILI in humans. Transgenic mouse strains (tgms) expressing HLA of serotype HLA-DR7 (HLA-DRB1*0701, -DRA*0102), and HLA-DQ2 (HLA-DQB1*0202,-DQA1*0201) were created. These two lines were crossed with a human (h)CD4 transgenic line, generating the two tgms DR7xhCD4 and DQ2xhCD4. To investigate whether the DILI effects observed in humans could be reproduced in tgms, the mice were treated for 28 days with ximelagatran. Results revealed no signs of DILI when biomarkers for liver toxicity were measured and histopathology was evaluated. In the ximelagatran case, presence of relevant HLA-expression in a pre-clinical model did not fulfil the prerequisite for reproducing DILI observed in patients. Nonetheless, for the first time an HLA-transgenic mouse model has been investigated for use in HLA-associated DILI induced by a low molecular weight compound. This study shows that mimicking of genetic susceptibility, expressed as DILI-associated HLA-types in mice, is not sufficient for reproducing the complex pathogenesis leading to DILI in man.
Asunto(s)
Azetidinas/toxicidad , Bencilaminas/toxicidad , Enfermedad Hepática Inducida por Sustancias y Drogas , Modelos Animales de Enfermedad , Antígenos HLA-DQ , Antígeno HLA-DR7 , Animales , Línea Celular , Enfermedad Hepática Inducida por Sustancias y Drogas/inmunología , Femenino , Antígenos HLA-DQ/genética , Antígenos HLA-DQ/metabolismo , Antígeno HLA-DR7/genética , Antígeno HLA-DR7/metabolismo , Humanos , Linfocitos/metabolismo , Masculino , Ratones Endogámicos C57BL , Ratones Transgénicos , Simulación del Acoplamiento Molecular , FenotipoRESUMEN
BACKGROUND: Following human mercury (Hg) exposure, the metal accumulates in considerable concentrations in kidney, liver, and brain. Although the toxicokinetics of Hg have been studied extensively, factors responsible for interindividual variation in humans are largely unknown. Differences in accumulation of renal Hg between inbred mouse strains suggest a genetic interstrain variation regulating retention or/and excretion of Hg. A.SW, DBA/2 and BALB/C mouse strains accumulate higher amounts of Hg than B10.S. OBJECTIVES: We aimed to find candidate genes associated with regulation of renal Hg concentrations. METHODS: A.SW, B10.S and their F1 and F2 offspring were exposed for 6 weeks to 2.0 mg Hg/L drinking water. Genotyping with microsatellites was conducted on 84 F2 mice for genome-wide scanning with ion pair reverse-phase high-performance liquid chromatography (IP RP HPLC). Quantitative trait loci (QTL) were established. Denaturing HPLC was used to detect single nucleotide polymorphisms for haplotyping and fine mapping in 184 and 32 F2 mice, respectively. Candidate genes (Pprc1, Btrc and Nfkb2) verified by fine mapping and QTL were further investigated by real-time polymerase chain reaction. Genes enhanced by Pprc1 (Nrf1 and Nrf2) were included for gene expression analysis. RESULTS: Renal Hg concentrations differed significantly between A.SW and B10.S mice and between males and females within each strain. QTL analysis showed a peak logarithm of odds ratio score 5.78 on chromosome 19 (p = 0.002). Haplotype and fine mapping associated the Hg accumulation with Pprc1, which encodes PGC-1-related coactivator (PRC), a coactivator for proteins involved in detoxification. Pprc1 and two genes coactivated by Pprc1 (Nrf1 and Nrf2) had significantly lower gene expression in the A.SW strain than in the B10.S strain. CONCLUSIONS: This study supports Pprc1 as a key regulator for renal Hg excretion. CITATION: Alkaissi H, Ekstrand J, Jawad A, Nielsen JB, Havarinasab S, Soderkvist P, Hultman P. 2016. Genome-wide association study to identify genes related to renal mercury concentrations in mice. Environ Health Perspect 124:920-926; http://dx.doi.org/10.1289/ehp.1409284.
Asunto(s)
Estudio de Asociación del Genoma Completo , Riñón/metabolismo , Mercurio/toxicidad , Animales , Expresión Génica/efectos de los fármacos , Genotipo , Haplotipos , Humanos , Riñón/efectos de los fármacos , Ratones , Ratones Endogámicos BALB C , Polimorfismo de Nucleótido Simple , Sitios de Carácter Cuantitativo , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Based on in vitro studies and short-term in vivo studies, all mercurials were for a long time considered as prototypic immunosuppressive substances. Recent studies have confirmed that organic mercurials such as methyl mercury (MeHg) and ethyl mercury (EtHg) are much more potent immunosuppressors than inorganic mercury (Hg). However, Hg interacts with the immune system in the presence of a susceptible genotype to cause immunostimulation, antinucleolar antibodies targeting fibrillarin, and systemic immune-complex (IC) deposits, a syndrome called Hg-induced autoimmunity (HgIA). Recent studies in mice with a susceptible genotype has revealed that the immunosuppressive effect of MeHg and EtHg will within 1-3 weeks be superseded by immunostimulation causing an HgIA-like syndrome. At equimolar doses of Hg, MeHg has the weakest immunostimulating, autoimmunogen, and IC-inducing effect, while the effect of thimerosal is similar to that of inorganic mercury. The immunosuppression is caused by the organic mercurials per se. Since they undergo rapid transformation to inorganic Hg, studies are being undertaken to delineate the importance of the organic substances per se and the newly formed inorganic Hg for induction of autoimmunity.
Asunto(s)
Enfermedades Autoinmunes , Mercurio/farmacología , Compuestos Organomercuriales/farmacología , Animales , Enfermedades Autoinmunes/inducido químicamente , Enfermedades Autoinmunes/inmunología , Enfermedades Autoinmunes/metabolismo , RatonesRESUMEN
Systemic lupus erythematosus is a multigenic disorder of unknown etiology. To investigate the role of specific genes in lupus, we have examined the effects of single gene deletions on mercury-induced autoimmunity. Deficiency of certain genes abrogated induction of autoimmunity, while absence of others had little effect. The most interesting observations were obtained with genes related to interferon-gamma. Genes involved in upregulation of IFN-gamma expression did not significantly influence autoimmunity whereas absence of IFN-gamma or IFN-gamma receptor led to greatly reduced autoantibody responses and immunopathology. Absence of IRF-1, a gene expressed in response to IFN-gamma, resulted in selective retention of anti-chromatin autoantibodies demonstrating that specific defects in signaling pathways and gene expression subsequent to IFN-gamma/IFN-gamma receptor interaction influence specific disease parameters. These studies show that single gene deletions can have various outcomes ranging from no effect, suppression of one or more features of disease, to suppression of all features of disease, and that all three outcomes can be observed in the IFN-gamma pathway. IFN-gamma influences the expression and function of other lupus relevant genes such as IL-6 and beta2microglobulin, therefore the effects of these gene deletions on disease expression may also reflect responses downstream of IFN-gamma function.
Asunto(s)
Enfermedades Autoinmunes/genética , Enfermedades Autoinmunes/inmunología , Animales , Enfermedades Autoinmunes/inducido químicamente , Enfermedades Autoinmunes/etiología , Lupus Eritematoso Sistémico/inducido químicamente , Lupus Eritematoso Sistémico/etiología , Lupus Eritematoso Sistémico/genética , Lupus Eritematoso Sistémico/inmunologíaRESUMEN
Methylmercury (MeHg) is a common environmental pollutant due to both natural and anthropogenic sources. Although the central nervous system (CNS) is considered the critical organ for the toxic effect of MeHg, it has recently been suggested that the immune system might be at least as sensitive as the CNS. We have examined the effects of MeHg on the immune system in genetically metal-susceptible mice. Subcutaneous (sc) injections of 2 mg MeHg/kg body weight (bw) every third day (internal dose ca. 540 microg Hg/kg bw/day) to A.SW mice of the H-2(s) haplotype, caused during the first week a 47 and 9% reduction of B- and T-cells, respectively, which indicates immunosuppression. Subsequently, an autoimmune syndrome developed which shared certain features with the syndrome induced by inorganic mercury in H-2(s) mice, including antibodies targeting the 34 kDa nucleolar protein fibrillarin, increased expression of IL-4 mRNA, increase of Th2-type of immunoglobulins (IgE and IgG1), and increased MHC class II expression on B-cells. However, the response using MeHg was attenuated compared with even lower doses of Hg in the form of inorganic mercury, and specifically lacked the increased expression of IL-2 and IFN-gamma mRNA, the polyclonal B-cell activation (PBA), and the systemic immune-complex (IC) deposits which are induced by inorganic mercury. Increasing the dose of MeHg increased the titre of anti-nucleolar antibodies and shortened the induction time, but did not lead to stronger immunostimulation or systemic IC-deposits. The kidney and liver selectively accumulated MeHg, while the blood, spleen and lymph nodes showed lower levels of MeHg. The accumulation of MeHg and Hg(2+) increased throughout the 30-day period. The fraction of Hg(2+) in the kidney varied between 4 and 22%, and the lymph nodes showed a maximum of 30% Hg(2+). We conclude first that MeHg has quantitatively different effect on the immune system compared with inorganic mercury, and secondly that an initial immunosuppression induced by a xenobiotic does not preclude subsequent immunostimulation and autoimmunity.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Contaminantes Ambientales/toxicidad , Inmunosupresores/toxicidad , Compuestos de Metilmercurio/toxicidad , Animales , Anticuerpos Antinucleares/sangre , Complejo Antígeno-Anticuerpo/metabolismo , Enfermedades Autoinmunes/inducido químicamente , Enfermedades Autoinmunes/genética , Linfocitos B/efectos de los fármacos , Linfocitos B/inmunología , Linfocitos B/metabolismo , Recuento de Células , Citocinas/biosíntesis , Relación Dosis-Respuesta a Droga , Contaminantes Ambientales/farmacocinética , Predisposición Genética a la Enfermedad , Inmunoglobulinas/sangre , Inmunosupresores/farmacocinética , Ganglios Linfáticos/efectos de los fármacos , Ganglios Linfáticos/metabolismo , Compuestos de Metilmercurio/farmacocinética , Ratones , Proyectos Piloto , Bazo/efectos de los fármacos , Bazo/inmunología , Bazo/metabolismo , Síndrome , Linfocitos T/efectos de los fármacosRESUMEN
A complete undergraduate medical programme in Linköping started 1986. The curriculum was innovative applying problem-based learning, community-orientation, and multi-professional training. After almost 20 years, a revision is implemented to vitalise the original educational principles. A curriculum committee coordinates seven multi-disciplinary theme-groups, mainly based on organ systems, responsible for planning and implementation of their parts during the whole curriculum. Critical appraisal, professional development, and population health are strengthened. Problem based learning is improved by using web-based scenarios and information technology. Phase I (2 semesters) focuses on basic concepts in basic science in relevant contexts, and Phase II (3 semesters) on normal structure, pathophysiology, diagnostic methods, and treatment. Phase III starts with a semester for a student research project and an elective period. The following five semesters deal with clinical medicine in hospitals and health centres with clerkships in four week periods changing with two week theoretical blocks related to the themes.