RESUMEN
Passiflora alata Curtis (P. alata) leaves have anti-inflammatory properties; the present study aimed to investigate the anti-diabetogenic properties of P. alata aqueous leaf extract. HPLC analysis identified the phenolic compounds catechin, epicatechin and rutin. The aqueous extract was administered for 30weeks to non-obese diabetic (NOD) mice presenting a decrease of 28.6% in diabetes incidence and the number of inflammatory cells in pancreatic islets, when compared with the control group (water). The P. alata group presented an antioxidant effect and decreased lipid peroxidation in the serum of NOD mice. Increased numbers of insulin-positive cells were also observed in the pancreatic islets of the treated group. The diabetic group exhibited higher levels in the glucose tolerance test and glycemic index, in comparison to the P. alata-treated group and non-diabetic control BALB/c mice. In addition, the P. alata extract reduced the percentage and the proliferation index of NOD mice lymphocytes submitted to in vitro dose/response mitogenic stimulation assays. These results suggest that the aqueous extract of P. alata has anti-inflammatory properties, contributing to the protection of beta cells in pancreatic islets in NOD mice, and presents potential for use a supporting approach to treat type 1 diabetes.
Asunto(s)
Antiinflamatorios/uso terapéutico , Antioxidantes/uso terapéutico , Diabetes Mellitus Tipo 1/tratamiento farmacológico , Células Secretoras de Insulina/efectos de los fármacos , Passiflora/inmunología , Extractos Vegetales/uso terapéutico , Animales , Células Cultivadas , Citoprotección , Femenino , Humanos , Insulina/sangre , Células Secretoras de Insulina/fisiología , Peroxidación de Lípido/efectos de los fármacos , Activación de Linfocitos/efectos de los fármacos , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos NOD , Hojas de la PlantaRESUMEN
Beta cell destruction in type 1 diabetes (TID) is associated with cellular oxidative stress and mitochondrial pathway of cell death. The aim of this study was to determine whether oxidative stress and mitochondrial dysfunction are present in T1D model (non-obese diabetic mouse, NOD) and if they are related to the stages of disease development. NOD mice were studied at three stages: non-diabetic, pre-diabetic, and diabetic and compared with age-matched Balb/c mice. Mitochondria respiration rates measured at phosphorylating and resting states in liver and soleus biopsies and in isolated liver mitochondria were similar in NOD and Balb/c mice at the three disease stages. However, NOD liver mitochondria were more susceptible to calcium-induced mitochondrial permeability transition as determined by cyclosporine-A-sensitive swelling and by decreased calcium retention capacity in all three stages of diabetes development. Mitochondria H2O2 production rate was higher in non-diabetic, but unaltered in pre-diabetic and diabetic NOD mice. The global cell reactive oxygen species (ROS), but not specific mitochondria ROS production, was significantly increased in NOD lymphomononuclear and stem cells in all disease stages. In addition, marked elevated rates of 2',7'-dichlorodihydrofluorescein (H2DCF) oxidation were observed in pancreatic islets from non-diabetic NOD mice. Using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) and lipidomic approach, we identified oxidized lipid markers in NOD liver mitochondria for each disease stage, most of them being derivatives of diacylglycerols and phospholipids. These results suggest that the cellular oxidative stress precedes the establishment of diabetes and may be the cause of mitochondrial dysfunction that is involved in beta cell death.
Asunto(s)
Enfermedades Autoinmunes/metabolismo , Diabetes Mellitus Experimental/metabolismo , Mitocondrias/metabolismo , Proteínas de Transporte de Membrana Mitocondrial/metabolismo , Estrés Oxidativo , Animales , Femenino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos NOD , Poro de Transición de la Permeabilidad Mitocondrial , Permeabilidad , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Here, we report the occurrence of leukocytoclastic vasculitis as an outcome of type III allergy to insulin in a patient with type II diabetes mellitus. The diagnosis was made on the basis of anatomo-pathological examination of a skin biopsy.
Asunto(s)
Hipersensibilidad/complicaciones , Enfermedades del Complejo Inmune/complicaciones , Insulina/inmunología , Vasculitis Leucocitoclástica Cutánea/diagnóstico , Vasculitis Leucocitoclástica Cutánea/etiología , Adulto , Diabetes Mellitus Tipo 2/complicaciones , Diabetes Mellitus Tipo 2/inmunología , Humanos , MasculinoRESUMEN
Recently we reported that monocyte phagocytosis and chemotaxis are impaired in X-linked agammaglobulinaemia (XLA) and common variable immunodeficiency (CVI) patients. Few data exist on the in vivo expression of receptors for the constant region of immunoglobulin (IgG) (Fc gammaR) and complement receptors (CR) in these patients. The objective of this study was to investigate the expression of Fc gammaR and CR on monocytes from XLA and CVI patients and compare it to that of healthy controls. Whole blood samples were obtained from 10 patients with XLA, 12 with CVI and 18 healthy controls. Monocyte phenotype was determined by flow cytometry with gating on CD14+ cells. Surface expression of Fc gammaRI (CD64), Fc gammaRII (CD32) and Fc gammaRIII (CD16), CR1 (CD35) and CR3 (CD11b and CD18) was measured by determination of the proportion of CD14+ cells positive for each receptor and by receptor density. Compared to controls, a significantly higher percentage of CD16 and CD35+ monocytes from XLA (P = 0.002 and P = 0.007, respectively) were observed. The relative fluorescence intensity (RFI) expression of Fc gammaRII (CD32) and Fc gammaRIII (CD16) were significantly lower on CVI monocytes compared to controls (P = 0.001 and P = 0.035, respectively). XLA patients, who have a reduction of Bruton's tyrosine kinase (Btk), showed normal or increased percentages of monocytes expressing Fc gamma and complement receptors. CVI patients, who have normal expression of Btk, showed reduced expression of CD16 and CD32 on monocytes. Inefficient chemotaxis and phagocytosis, reported previously in XLA patients, could be due to defects of cytoplasmatic transduction mechanisms.
Asunto(s)
Agammaglobulinemia/inmunología , Inmunodeficiencia Variable Común/inmunología , Monocitos/inmunología , Receptores de Complemento/sangre , Receptores de IgG/sangre , Adolescente , Adulto , Agammaglobulinemia/genética , Antígenos CD/sangre , Niño , Preescolar , Femenino , Proteínas Ligadas a GPI , Enfermedades Genéticas Ligadas al Cromosoma X/inmunología , Humanos , Inmunofenotipificación , MasculinoRESUMEN
We have described previously the prophylactic and therapeutic effect of a DNA vaccine encoding the Mycobacterium leprae 65 kDa heat shock protein (DNA-HSP65) in experimental murine tuberculosis. However, the high homology of this protein to the corresponding mammalian 60 kDa heat shock protein (Hsp60), together with the CpG motifs in the plasmid vector, could trigger or exacerbate the development of autoimmune diseases. The non-obese diabetic (NOD) mouse develops insulin-dependent diabetes mellitus (IDDM) spontaneously as a consequence of an autoimmune process that leads to destruction of the insulin-producing beta cells of the pancreas. IDDM is characterized by increased T helper 1 (Th1) cell responses toward several autoantigens, including Hsp60, glutamic acid decarboxylase and insulin. In the present study, we evaluated the potential of DNA-HSP65 injection to modulate diabetes in NOD mice. Our results show that DNA-HSP65 or DNA empty vector had no diabetogenic effect and actually protected NOD mice against the development of severe diabetes. However, this effect was more pronounced in DNA-HSP65-injected mice. The protective effect of DNA-HSP65 injection was associated with a clear shift in the cellular infiltration pattern in the pancreas. This change included reduction of CD4(+) and CD8(+) T cells infiltration, appearance of CD25(+) cells influx and an increased staining for interleukin (IL)-10 in the islets. These results show that DNA-HSP65 can protect NOD mice against diabetes and can therefore be considered in the development of new immunotherapeutic strategies.
Asunto(s)
Diabetes Mellitus Tipo 1/prevención & control , Vacunas contra la Tuberculosis/inmunología , Animales , Anticuerpos Antibacterianos/biosíntesis , Autoantígenos/inmunología , Proteínas Bacterianas/inmunología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Chaperonina 60 , Chaperoninas/inmunología , Progresión de la Enfermedad , Inmunoglobulina G/biosíntesis , Interleucina-10/metabolismo , Subunidad alfa del Receptor de Interleucina-2/análisis , Islotes Pancreáticos/inmunología , Ratones , Ratones Endogámicos NOD , Factor de Necrosis Tumoral alfa/metabolismo , Vacunas de ADN/inmunologíaRESUMEN
BACKGROUND: Frequent exposure to latex causes various reactions such as respiratory symptoms and anaphylactic shock. In these cases, proteins found in natural latex are responsible for the serious systemic antilatex-mediated immediate hypersensitive reactions. METHODS: Cross-sectional descriptive survey focusing on 96 Brazilian health care workers (HCW) in the neonatal intensive care unit at CAISM, State University of Campinas UNICAMP, Brazil. All subjects were interviewed, donated blood samples for the latex-specific immunoglobulin E measurement and underwent the skin prick test (SPT) with an antigen extracted from latex gloves. RESULTS: The prevalence of latex positive SPT was 8%. There were eight SPT positive and only one serologic test was in agreement with the SPT. Overall, there was evidence of an association between the latex SPT and reported eczema (P = 0.01); food allergy (P = 0.009) with pineapple (P = 0.01). CONCLUSIONS: These results suggest that the identification of reactions of immediate hypersensitivity mediated by antilatex antibodies in HCW should be encouraged to prevent occupational exposure to latex products.
Asunto(s)
Hipersensibilidad al Látex/diagnóstico , Hipersensibilidad al Látex/epidemiología , Exposición Profesional/efectos adversos , Adulto , Alérgenos/farmacología , Antígenos/análisis , Brasil/epidemiología , Estudios Transversales , Femenino , Humanos , Recién Nacido , Unidades de Cuidado Intensivo Neonatal , Látex/inmunología , Masculino , Persona de Mediana Edad , Personal de Hospital , Prevalencia , Probabilidad , Medición de Riesgo , Pruebas Cutáneas , Estadísticas no ParamétricasRESUMEN
Affinity diacetylene liposomes were prepared with 10,12-tricosadiynoic acid and cardiolipin as the affinity ligand for anticardiolipin antibodies at a molar ratio of 80:20. Polymerization was carried out under UV irradiation, and the color transitions were monitored by visible absorption spectroscopy. Peaks at 635 nm (blue form), 540 nm (purple form), and 480 nm (red form) were observed as a function of time. These polymerized liposomes were used in a noncompetitive immunoassay for detection of anticardiolipin antibodies. Color changes were observed when reference serum containing specific immunoglobulin G, IgG, was added to polymerized liposome dispersions. The colorimetric signal due to IgG adsorption on the liposome surface was quantified as a colorimetric response defined as the change in percentage of blue color related to the initial percentage of blue color in the solutions. The colorimetric response was 10 times higher for specific IgG compared with nonspecific ones. These results suggest the unique potentialities of affinity diacetylene polymerized liposomes in the development of biosensors for diagnosis of autoimmune diseases.
Asunto(s)
Acetileno/química , Autoanticuerpos/análisis , Liposomas/síntesis química , Polímeros/síntesis química , Acetileno/análogos & derivados , Autoanticuerpos/inmunología , Cardiolipinas/química , Cardiolipinas/inmunología , Ácidos Grasos Insaturados/química , Ácidos Grasos Insaturados/inmunología , Inmunoglobulina G/análisis , Inmunoglobulina G/inmunología , Ligandos , Liposomas/química , Liposomas/inmunología , Estructura Molecular , Polímeros/química , Espectrofotometría Ultravioleta , Factores de TiempoRESUMEN
Tumor necrosis factor-alpha (TNF-alpha) is one of the most important proinflammatory cytokines which plays a central role in host defense and in the acute inflammatory response related to tissue injury. The major source of TNF-alpha are immune cells such as neutrophils and macrophages. We tested the hypothesis that pentoxifylline, a methylxanthine derivative, down-regulates proinflammatory cytokine expression during acute lung injury in rats. Male Wistar rats weighing 250 to 450 g were anesthetized ip with 50 mg/kg sodium thiopental and randomly divided into three groups: group 1 (N = 7): tidal volume (V T) = 7 ml/kg, respiratory rate (RR) = 50 breaths/min and normal saline infusion; group 2 (N = 7): V T = 42 ml/kg, RR = 9 breaths/min and normal saline infusion; group 3 (N = 7): V T = 42 ml/kg, RR = 9 breaths/min and pentoxifylline infusion. The animals were ventilated with an inspired oxygen fraction of 1.0, a positive end-expiratory pressure of 3 cmH2O, and normal saline or pentoxifylline injected into the left femoral vein. The mRNA of TNF-alpha rapidly increased in the lung tissue within 180 min of ventilation with a higher V T with normal saline infusion. The concentrations of inflammatory mediators were decreased in plasma and bronchoalveolar lavage (BAL) in the presence of higher V T with pentoxifylline infusion (TNF-alpha: plasma, 102.2 ± 90.9 and BAL, 118.2 ± 82.1; IL-1ß: plasma, 45.2 ± 42.7 and BAL, 50.2 ± 34.9, P < 0.05). We conclude that TNF-alpha produced by neutrophil influx may function as an alert signal in host defense to induce production of other inflammatory mediators
Asunto(s)
Animales , Masculino , Ratas , Interleucina-1 , Pentoxifilina , Inhibidores de Fosfodiesterasa , Síndrome de Dificultad Respiratoria/tratamiento farmacológico , Factor de Necrosis Tumoral alfa , Análisis de los Gases de la Sangre , Lavado Broncoalveolar , Mediciones del Volumen Pulmonar , Ratas Wistar , Respiración Artificial , Volumen de Ventilación Pulmonar , Factor de Necrosis Tumoral alfaRESUMEN
Tumor necrosis factor-alpha (TNF-alpha) is one of the most important proinflammatory cytokines which plays a central role in host defense and in the acute inflammatory response related to tissue injury. The major source of TNF-alpha are immune cells such as neutrophils and macrophages. We tested the hypothesis that pentoxifylline, a methylxanthine derivative, down-regulates proinflammatory cytokine expression during acute lung injury in rats. Male Wistar rats weighing 250 to 450 g were anesthetized ip with 50 mg/kg sodium thiopental and randomly divided into three groups: group 1 (N = 7): tidal volume (V T) = 7 ml/kg, respiratory rate (RR) = 50 breaths/min and normal saline infusion; group 2 (N = 7): V T = 42 ml/kg, RR = 9 breaths/min and normal saline infusion; group 3 (N = 7): V T = 42 ml/kg, RR = 9 breaths/min and pentoxifylline infusion. The animals were ventilated with an inspired oxygen fraction of 1.0, a positive end-expiratory pressure of 3 cmH2O, and normal saline or pentoxifylline injected into the left femoral vein. The mRNA of TNF-alpha rapidly increased in the lung tissue within 180 min of ventilation with a higher V T with normal saline infusion. The concentrations of inflammatory mediators were decreased in plasma and bronchoalveolar lavage (BAL) in the presence of higher V T with pentoxifylline infusion (TNF-alpha: plasma, 102.2+/-90.9 and BAL, 118.2+/-82.1; IL-1 : plasma, 45.2+/-42.7 and BAL, 50.2+/-34.9, P < 0.05). We conclude that TNF-alpha produced by neutrophil influx may function as an alert signal in host defense to induce production of other inflammatory mediators.
Asunto(s)
Interleucina-1/sangre , Pentoxifilina/farmacología , Inhibidores de Fosfodiesterasa/farmacología , Síndrome de Dificultad Respiratoria/tratamiento farmacológico , Factor de Necrosis Tumoral alfa/análisis , Animales , Análisis de los Gases de la Sangre , Lavado Broncoalveolar , Mediciones del Volumen Pulmonar , Masculino , Ratas , Ratas Wistar , Respiración Artificial , Volumen de Ventilación Pulmonar , Factor de Necrosis Tumoral alfa/efectos de los fármacosRESUMEN
The role of the basement membrane as an antigenic structure in autoimmune diseases is controversial. To determine possible structural changes in the endocrine pancreatic basement membrane (PBM) in autoimmune diabetes, we studied the expression of laminin in the islets of 42 NOD mice, aged between 4 to 42 weeks, as an animal model of spontaneous diabetes. Insular lymphocytic inflammatory infiltration of variable intensity was present in 24 of these mice. An immunohistochemical staining using the streptavidin-biotin-peroxidase technique was performed on paraffin-embedded tissue sections, with a polyclonal antilaminin antibody. Staining for laminin was restricted to the basement membrane. In islets with no inflammatory infiltration, laminin was observed as a thin, continuous and uniform brown layer, covering the pericapsular basement membrane of the islets and their capillaries. The continuity of the PBM was lost in the islets with insulitis and the immunostaining showed clearcut interruption and destruction, particularly when the islets were in contact with inflammatory infiltrate. Our findings suggest that the loss of integrity of the PBM in islets with inflammatory infiltrate could facilitate antigenic exposure contributing towards the start o f autoimmune DM in NODmice.
Asunto(s)
Membrana Basal/patología , Diabetes Mellitus Tipo 1/patología , Islotes Pancreáticos/patología , Ratones Endogámicos NOD , Pancreatitis/patología , Animales , Membrana Basal/metabolismo , Diabetes Mellitus Tipo 1/complicaciones , Diabetes Mellitus Tipo 1/metabolismo , Femenino , Técnicas para Inmunoenzimas , Islotes Pancreáticos/metabolismo , Laminina/metabolismo , Masculino , Ratones , Pancreatitis/etiología , Pancreatitis/metabolismoRESUMEN
Insulin-dependent diabetes mellitus is caused by autoimmune destruction of pancreatic beta cells. Non-obese diabetic (NOD) mice spontaneously develop diabetes similar to the human disease. Cytokines produced by islet-infiltrating mononuclear cells may be directly cytotoxic and can be involved in islet destruction coordinated by CD4+ and CD8+ cells. We utilized a semiquantitative RT-PCR assay to analyze in vitro the mRNA expression of TNF-alpha and IFN-gamma cytokine genes in isolated islets (N = 100) and spleen cells (5 x 10(5) cells) from female NOD mice during the development of diabetes and from female CBA-j mice as a related control strain that does not develop diabetes. Cytokine mRNAs were measured at 2, 4, 8, 14 and 28 weeks of age from the onset of insulitis to the development of overt diabetes. An increase in IFN-gamma expression in islets was observed for females aged 28 weeks (149 +/- 29 arbitrary units (AU), P<0.05, Student t-test) with advanced destructive insulitis when compared with CBA-j mice, while TNF-alpha was expressed in both NOD and CBA-j female islets at the same level at all ages studied. In contrast, TNF-alpha in spleen was expressed at higher levels in NOD females at 14 weeks (99 +/- 8 AU, P<0.05) and 28 weeks (144 +/- 17 AU, P<0.05) of age when compared to CBA-j mice. The data suggest that IFN-gamma and TNF-alpha expression in pancreatic islets of female NOD mice is associated with beta cell destruction and overt diabetes.
Asunto(s)
Diabetes Mellitus Tipo 1/metabolismo , Interferón gamma/metabolismo , Islotes Pancreáticos/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Factores de Edad , Animales , Femenino , Expresión Génica , Interferón gamma/genética , Cinética , Ratones , Ratones Endogámicos CBA , Ratones Endogámicos NOD , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Bazo/citología , Bazo/metabolismo , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
Insulin-dependent diabetes mellitus is caused by autoimmune destruction of pancreatic ß cells. Non-obese diabetic (NOD) mice spontaneously develop diabetes similar to the human disease. Cytokines produced by islet-infiltrating mononuclear cells may be directly cytotoxic and can be involved in islet destruction coordinated by CD4+ and CD8+ cells. We utilized a semiquantitative RT-PCR assay to analyze in vitro the mRNA expression of TNF-alpha and IFN-gamma cytokine genes in isolated islets (N = 100) and spleen cells (5 x 10(5) cells) from female NOD mice during the development of diabetes and from female CBA-j mice as a related control strain that does not develop diabetes. Cytokine mRNAs were measured at 2, 4, 8, 14 and 28 weeks of age from the onset of insulitis to the development of overt diabetes. An increase in IFN-gamma expression in islets was observed for females aged 28 weeks (149 ± 29 arbitrary units (AU), P<0.05, Student t-test) with advanced destructive insulitis when compared with CBA-j mice, while TNF-alpha was expressed in both NOD and CBA-j female islets at the same level at all ages studied. In contrast, TNF-alpha in spleen was expressed at higher levels in NOD females at 14 weeks (99 ± 8 AU, P<0.05) and 28 weeks (144 ± 17 AU, P<0.05) of age when compared to CBA-j mice. The data suggest that IFN-gamma and TNF-alpha expression in pancreatic islets of female NOD mice is associated with ß cell destruction and overt diabetes
Asunto(s)
Animales , Femenino , Ratones , Diabetes Mellitus Tipo 1 , Interferón gamma , Islotes Pancreáticos , Factor de Necrosis Tumoral alfa , Factores de Edad , Expresión Génica , Interferón gamma , Cinética , Ratones Endogámicos NOD , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , ARN Mensajero , Bazo , Factor de Necrosis Tumoral alfaRESUMEN
Nonobese diabetic (NOD) mice and a derived strain, NOD.H.2h4, have been used as a model for experimental spontaneous thyroiditis and thyroiditis induced by iodide excess after a goiter-inducing period. Some authors have proposed that iodide, given after methimazole or propylthiouracil, is capable of inducing apoptosis in thyroid cells and that anti-thyroid drugs can modulate the expression of apoptosis components such as Fas and its ligand (Fas-L). Here we evaluated the effect of potassium iodide (20 microg/animal for 4 days, i.p.) given to NOD mice at the 10th week of life after exposure to methimazole (1 mg/ml) in drinking water from the 4th to the 10th week of life. Fas, Fas-L and Bcl-w expression were analyzed semiquantitatively by RT-PCR immediately after potassium iodide administration (group MI44D) or at week 32 (MI32S). Control groups were added at 10 (C10) and 32 weeks (C32), as well as a group that received only methimazole (CM10). An increase in the expression of Fas-L and Bcl-w (P<0.01, ANOVA) was observed in animals of group MI44D, while Fas was expressed at higher levels (P = 0.02) in group C32 (72.89 +/- 47.09 arbitrary units) when compared to group C10 (10.8 +/- 8.55 arbitrary units). Thus, the analysis of Fas-L and Bcl-w expression in the MI44D group and Fas in group C32 allowed us to detect two different patterns of expression of these apoptosis components in thyroid tissue of NOD mice.
Asunto(s)
Apoptosis/efectos de los fármacos , Yoduro de Potasio/farmacología , ARN Mensajero/efectos de los fármacos , Glándula Tiroides/efectos de los fármacos , Tiroiditis/inmunología , Receptor fas/efectos de los fármacos , Animales , Antitiroideos/farmacología , Apoptosis/fisiología , Modelos Animales de Enfermedad , Electroforesis , Proteína Ligando Fas , Expresión Génica , Masculino , Glicoproteínas de Membrana , Metimazol/farmacología , Ratones , Ratones Endogámicos NOD , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Receptor fas/análisisRESUMEN
Nonobese diabetic (NOD) mice and a derived strain, NOD.H.2h4, have been used as a model for experimental spontaneous thyroiditis and thyroiditis induced by iodide excess after a goiter-inducing period. Some authors have proposed that iodide, given after methimazole or propylthiouracil, is capable of inducing apoptosis in thyroid cells and that anti-thyroid drugs can modulate the expression of apoptosis components such as Fas and its ligand (Fas-L). Here we evaluated the effect of potassium iodide (20 æg/animal for 4 days, ip) given to NOD mice at the 10th week of life after exposure to methimazole (1 mg/ml) in drinking water from the 4th to the 10th week of life. Fas, Fas-L and Bcl-w expression were analyzed semiquantitatively by RT-PCR immediately after potassium iodide administration (group MI44D) or at week 32 (MI32S). Control groups were added at 10 (C10) and 32 weeks (C32), as well as a group that received only methimazole (CM10). An increase in the expression of Fas-L and Bcl-w (P<0.01, ANOVA) was observed in animals of group MI44D, while Fas was expressed at higher levels (P = 0.02) in group C32 (72.89 ± 47.09 arbitrary units) when compared to group C10 (10.8 ± 8.55 arbitrary units). Thus, the analysis of Fas-L and Bcl-w expression in the MI44D group and Fas in group C32 allowed us to detect two different patterns of expression of these apoptosis components in thyroid tissue of NOD mice
Asunto(s)
Animales , Masculino , Ratones , Receptor fas , Apoptosis , Yoduro de Potasio , ARN Mensajero , Glándula Tiroides , Tiroiditis , Antitiroideos , Apoptosis , Modelos Animales de Enfermedad , Electroforesis , Expresión Génica , Glicoproteínas de Membrana , Metimazol , Ratones Endogámicos NOD , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Histidine was immobilized onto PEVA membrane to obtain an affinity support for human IgG removal from serum with a view to clinical apheresis for the treatment of autoimmune diseases. These membranes were able to remove in vitro several autoantibodies from the serum of SLE patients.
Asunto(s)
Filtración/métodos , Histidina/química , Inmunoglobulina G/inmunología , Inmunoglobulina G/aislamiento & purificación , Polivinilos/química , Aminoácidos/química , Anticuerpos/aislamiento & purificación , Humanos , Inmunoglobulina G/sangre , Lupus Eritematoso Sistémico/sangre , Lupus Eritematoso Sistémico/inmunología , Lupus Eritematoso Sistémico/terapiaRESUMEN
Studies concerning the antigenicity of thyroglobulin fragments allow the characterization of the epitopes but do not consider the role of heavier antigenic fragments that could result in vivo from the action of endoproteases. Here we assess the relative importance of the fragments obtained from thyroglobulin by limited proteolysis with trypsin and compare by immunoblotting their reactivity to serum from patients with autoimmune (Graves' disease and Hashimoto's thyroiditis) and non-autoimmune (subacute thyroiditis) disease. The results showed no difference in frequency of recognition of any peptide by sera from patients with autoimmune thyroiditis. In contrast, sera from patients with subacute thyroiditis reacted more frequently with a peptide of 80 kDa. These results suggest the presence of antibody subpopulations directed at fragments produced in vivo by enzymatic cleavage of thyroglobulin. This fragment and antibodies to it may represent markers for subacute thyroiditis.
Asunto(s)
Anticuerpos/inmunología , Enfermedad de Graves/inmunología , Fragmentos de Péptidos/inmunología , Tiroglobulina/inmunología , Tiroiditis Autoinmune/inmunología , Tiroiditis Subaguda/inmunología , HumanosRESUMEN
Studies concerning the antigenicity of thyroglobulin fragments allow the characterization of the epitopes but do not consider the role of heavier antigenic fragments that could result in vivo from the action of endoproteases. Here we assess the relative importance of the fragments obtained from thyroglobulin by limited proteolysis with trypsin and compare by immunoblotting their reactivity to serum from patients with autoimmune (Graves' disease and Hashimoto's thyroiditis) and non-autoimmune (subacute thyroiditis) disease. The results showed no difference in frequency of recognition of any peptide by sera from patients with autoimmune thyroiditis. In contrast, sera from patients with subacute thyroiditis reacted more frequently with a peptide of 80 kDa. These results suggest the presence of antibody subpopulations directed at fragments produced in vivo by enzymatic cleavage of thyroglobulin. This fragment and antibodies to it may represent markers for subacute thyroiditis.
Asunto(s)
Humanos , Anticuerpos , Enfermedad de Graves/inmunología , Fragmentos de Péptidos/inmunología , Tiroglobulina/inmunología , Tiroiditis Autoinmune/inmunología , Tiroiditis Subaguda/inmunología , Hidrólisis , Immunoblotting , TripsinaRESUMEN
The basement membrane as an antigenic structure in autoimmune diseases has been a matter of controversy. The purpose of our study was to determine possible structural changes in the follicular basement membrane (FBM) in thyroid autoimmune and non-autoimmune diseases. Immunohistochemical staining for collagen IV and laminin showed that the continuity of the FBM was preserved in toxic adenoma (three cases), atoxic multinodular goiter (nine cases) as well as in autoimmune disease. Integrity of the FBM was observed in all 11 cases of Graves' disease and Hashimoto's thyroiditis (seven cases) studied. In some instances, the FBM was thinned in areas of contact with inflammatory infiltrate. We conclude that the auto-antibodies, and possibly other factors present in autoimmune thyroid diseases, do not significantly alter the integrity of the FBM.