RESUMEN
OBJECTIVE: The purpose of this study is to evaluate the use of diffusion-weighted magnetic resonance imaging (DWMRI) in the assessment of graft rejection after liver transplantation (LT). METHODS: From June 2017 to January 2018, 32 patients were included in the study with a mean age of 52.3 years. All patients underwent LT. The DWMRI was performed using the apparent diffusion coefficient map and measuring the different b-values (b-400, b-600, b-800, and b-1000). These measurements were compared with the histopathology results. Statistical analysis included t test, analysis of variance, and area under the curve for receiver operating characteristic (ROC). RESULTS: There were 17 patients without rejection and 15 patients with liver graft rejection diagnosed by histopathology. The mean (SD) results between the nonrejection and rejection groups were as follows: b-400 = 1.568 (0.265) vs 1.519 (0.119) (P = .089), b-600 = 1.380 (0.181) vs 1.284 (0.106) (P = .039), b-800 = 1.262 (0.170) vs 1.170 (0.086) (P = .035), b-1000 = 1.109 (0.129) vs 1.098 (0.078) (P = .095); B-values × 10-3 mm2/s. Only b-600 (P = .04) and b-800 (P = .04) values have significant differences between the 2 groups. B-600 showed 90.48% sensitivity and 83.33% specificity (ROC area under the curve = 0.784; P < .001), and b-800 showed 90.38% sensitivity and 83.03% specificity (ROC area under the curve = 0.816; P < .001). The values obtained with the apparent diffusion coefficient in b-800 were clearly differentiated between the mild, moderate, and severe degrees of rejection (P < .001). CONCLUSION: Measurement of b-600 and b-800 values using DWMRI may be used for the diagnosis of graft rejection after LT.
Asunto(s)
Imagen de Difusión por Resonancia Magnética/métodos , Rechazo de Injerto/diagnóstico por imagen , Trasplante de Hígado/efectos adversos , Hígado/diagnóstico por imagen , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Curva ROC , Sensibilidad y EspecificidadRESUMEN
Cullin 4B (CUL4B), a member of the cullin protein family, is a scaffold protein of the CUL4B-RING-E3 ligase complex that ubiquitinates intracellular proteins.CUL4B's targets include cell cycle-regulated proteins and DNA replication-related molecules. In this study, we generated myeloid-specific Cul4b-deficient mice (Cul4b(f/y);LysM-Cre(KI/KI)) to investigate the influence of Cul4b deficiency on innate immunity, especially on the function of macrophages. Our results show that an intraperitoneal injection of lipopolysaccharide (LPS) led to a significant decrease in body weights and increased leukocyte infiltrates with increased chemokines in the peritoneal cavity of Cul4b(f/y);LysM-Cre(KI/KI) mice. However, the proinflammatory cytokines, IL-6 and TNF-α did not increase in LPS-injected Cul4b(f/y);LysM-Cre(KI/KI) mice. Furthermore, bone marrow-derived macrophages from Cul4b(f/y);LysM-Cre(KI/KI) mice secreted higher levels of chemokines but lower levels of TNF-α and IL-6 upon LPS stimulation. Of note, increased proliferation of Cul4b-deficient macrophages was also observed. These results show that myeloid-specific Cul4b deficiency worsens LPS-induced peritonitis. In addition, Cul4b deficiency leads to enhanced DNA replication and proliferation, increased production of chemokines but a decreased production of proinflammatory cytokines of macrophages. Our data highlight a new role of cullin family, CUL4B, in the immune system.
Asunto(s)
Proteínas Cullin/inmunología , Inmunidad Innata/inmunología , Macrófagos/inmunología , Peritonitis/inmunología , Animales , Peso Corporal/inmunología , Células de la Médula Ósea/inmunología , Células de la Médula Ósea/metabolismo , Ciclo Celular/genética , Ciclo Celular/inmunología , Línea Celular Tumoral , Proliferación Celular/genética , Quimiocinas/inmunología , Quimiocinas/metabolismo , Proteínas Cullin/genética , Proteínas Cullin/metabolismo , Expresión Génica/inmunología , Inmunidad Innata/genética , Inmunoensayo , Interleucina-6/genética , Interleucina-6/inmunología , Interleucina-6/metabolismo , Leucocitos/inmunología , Leucocitos/metabolismo , Lipopolisacáridos , Macrófagos/metabolismo , Ratones , Ratones Noqueados , Peritonitis/inducido químicamente , Peritonitis/genética , Fagocitosis/genética , Fagocitosis/inmunología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunología , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Although primary biliary cirrhosis (PBC) is considered a model autoimmune disease, it has not responded therapeutically to traditional immunosuppressive agents. In addition, PBC may recur following liver transplantation, despite the absence of major histocompatibility complex (MHC) matching, in sharp contrast to the well-known paradigm of MHC restriction. We have suggested previously that invariant natural killer T (iNK T) cells are critical to the initiation of PBC. In this study we have taken advantage of our ability to induce autoimmune cholangitis with 2-octynoic acid, a common component of cosmetics, conjugated to bovine serum albumin (2-OA-BSA), and studied the natural history of pathology in mice genetically deleted for CD4 or CD8 following immunization with 2-OA-BSA in the presence or absence of α-galactosylceramide (α-GalCer). In particular, we address whether autoimmune cholangitis can be induced in the absence of traditional CD4 and CD8 responses. We report herein that CD4 and CD8 knock-out mice immunized with 2-OA-BSA/PBS or 2-OA-BSA/α-GalCer develop anti-mitochondrial antibodies (AMAs), portal infiltrates and fibrosis. Indeed, our data suggest that the innate immunity is critical for immunopathology and that the pathology is exacerbated in the presence of α-GalCer. In conclusion, these data provide not only an explanation for the recurrence of PBC following liver transplantation in the absence of MHC compatibility, but also suggest that effective therapies for PBC must include blocking of both innate and adaptive pathways.
Asunto(s)
Enfermedades Autoinmunes/inmunología , Colangitis/inmunología , Inmunidad Innata , Animales , Autoanticuerpos/sangre , Autoanticuerpos/inmunología , Enfermedades Autoinmunes/inducido químicamente , Enfermedades Autoinmunes/genética , Antígenos CD4/genética , Antígenos CD4/inmunología , Antígenos CD8/genética , Antígenos CD8/inmunología , Colangitis/inducido químicamente , Colangitis/genética , Acetiltransferasa de Residuos Dihidrolipoil-Lisina/inmunología , Modelos Animales de Enfermedad , Ácidos Grasos Monoinsaturados/efectos adversos , Ácidos Grasos Monoinsaturados/inmunología , Femenino , Galactosilceramidas/administración & dosificación , Galactosilceramidas/efectos adversos , Hígado/inmunología , Hígado/metabolismo , Hígado/patología , Cirrosis Hepática Biliar/inmunología , Ratones , Ratones Noqueados , Proteínas Mitocondriales/inmunología , Albúmina Sérica Bovina/efectos adversos , Albúmina Sérica Bovina/inmunología , Xenobióticos/efectos adversosRESUMEN
BACKGROUND: Henoch-Schönlein purpura (HSP) is a common IgA-mediated vasculitis in children. The antigenic target for IgA is to be determined. OBJECTIVE: To test whether ß2-glycoprotein I (ß2GPI) is an antigenic target for IgA in childhood HSP, and to evaluate the clinical implications and pathogenic role of such IgA autoantibodies. METHODS: The reactivity of patients' plasma samples and purified polyclonal IgA with ß2GPI, ß2GPI-derived peptides and endothelial cells was tested by enzyme-linked immunosorbent assay. The association between clinical manifestations and IgA anti-ß2GPI antibodies was also analysed. Finally, IgA-mediated cytotoxicity on endothelial cells was further evaluated. RESULTS: At the acute stage, patients with HSP had significantly higher plasma levels of IgA antibodies against ß2GPI than healthy controls [reference units (RU) 1.14 ± 0.8 vs. 0.42 ± 0.24, P < 0.001]. IgA anti-ß2GPI antibodies were associated with the presence of joint manifestations (with vs. without joint involvement, 1.15 ± 0.64 vs. 0.85 ± 0.47, P = 0.0341) and heavy proteinuria (with vs. without heavy proteinuria, 2.09 ± 2.02 vs. 1.04 ± 0.62, P = 0.0028). Polyclonal IgA from plasma samples positive for IgA anti-ß2GPI antibodies bound well not only to ß2GPI with Kd values < 10(-5) mol L(-1), but also to some ß2GPI-dereived linear peptides (P3, P5, P7, P11 and P12). Moreover, ß2GPI-reactive polyclonal IgA also bound well to endothelial cells and induced complement-dependent cell lysis. CONCLUSION: These findings reveal the clinical and pathogenic relevance of IgA anti-ß2GPI antibodies in childhood HSP and suggest that ß2GPI may be an important autoantigen for HSP.
Asunto(s)
Vasculitis por IgA/inmunología , Inmunoglobulina A/sangre , beta 2 Glicoproteína I/inmunología , Adolescente , Estudios de Casos y Controles , Niño , Preescolar , Células Endoteliales/inmunología , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inmunoglobulina A/inmunología , MasculinoRESUMEN
Vigorous treatment of aggressive digital papillary adenocarcinoma (ADPA), including amputation, has been recommended by most authors, but the appropriateness and effectiveness of excision as an alternative to amputation has not been systematically evaluated. To evaluate the appropriateness and effectiveness of excision as an alternative to amputation in the treatment of ADPA, we reviewed the clinical presentations, treatments and patient outcomes presented in case reports on ADPA available on Ovid MEDLINE. We also assessed the results of immunohistochemical staining for proliferation markers in one patient in order to explain the nonaggressive nature of ADPA noted in that patient. Except for the duration of lesions, there was no significant difference in clinical outcome between the excision and amputation groups. We also found that p63 may be a useful marker for distinguishing primary ADPA from metastatic adenocarcinomas. In addition, the intensity of Ki67 expression in tumour cells may be a marker of aggressive behaviour and thus be helpful in therapeutic decision-making. Wide excision with or without sentinel lymph-node biopsy is a feasible alternative to amputation. It should be considered in patients who present with a long-standing history of ADPA without evidence of underlying bone invasion or distant metastasis and with low-intensity expression of proliferation markers.
Asunto(s)
Adenocarcinoma Papilar , Amputación Quirúrgica , Neoplasias Cutáneas , Adenocarcinoma Papilar/patología , Adenocarcinoma Papilar/cirugía , Dedos/cirugía , Humanos , Neoplasias Cutáneas/patología , Neoplasias Cutáneas/cirugía , Factores de Tiempo , Dedos del Pie/cirugíaRESUMEN
BACKGROUND: Amphotericin B (AmB) has a discordant influence on epirubicin (4'-epidoxorubicin) cytotoxicity in hepatocellular carcinoma (HCC). This indicates that the cellular function of HCC may be significantly influenced by AmB. Whether the influence of AmB on HCC has any possibility to influence cancer growth has not been studied. This study was to try and clarify this issue. MATERIALS AND METHODS: Two HCC cell lines including one without augmentation of the epirubicin cytotoxicity by AmB (cell line A; HCC24/KMUH) and one with this effect (cell line B; HCC38/KMUH) were studied by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and whole human genome microarray (experimental group: 2.5 microg mL(-1) AmB). RESULTS: Differential expressions of genes induced by AmB in two cell lines had no influence on cell proliferation as determined by MTT assay. Only cell line B showed up-regulation of genes related to oxidative stress, acute phase reaction, cytokine-cytokine receptor interaction and complement and coagulation cascades. Among the chemokine genes up-regulated by AmB, five genes (CCL2, CXCL1, CXCL5, CXCL6, IL8) were angiogenic. Cell line B also showed up-regulation of one angiogenic C10orf10 gene and down-regulation of one angiostatic chemokine gene (CXCL10). Up- or down-regulation of other genes in cell line A and B did not show any evidence to promote angiogenesis. CONCLUSION: AmB has the capacity to concomitantly up-regulate angiogenic genes in HCC cells susceptible to AmB-induced oxidative stress.
Asunto(s)
Anfotericina B/uso terapéutico , Antifúngicos/uso terapéutico , Carcinoma Hepatocelular/tratamiento farmacológico , Regulación Neoplásica de la Expresión Génica/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacos , Carcinoma Hepatocelular/genética , Humanos , Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Células Tumorales Cultivadas , Regulación hacia Arriba/genéticaRESUMEN
Dendritic cells (DCs) can induce both tolergenic as well as effective immune responses in the lung. Pulmonary DCs producing interleukin (IL)-10 mediated tolerance induced by respiratory exposure to antigen. IL-10 is an important immunosuppressive cytokine, which inhibits maturation and function of DC. To assess whether IL-10 producing DCs can exert the tolergenic effect through the differentiation of regulatory T cells, bone marrow derived DCs were genetically modified by IL-10 expressing adenovirus. IL-10 gene modified DCs (Ad-IL-10-DC) displayed a characteristic phenotype of immature DCs. Here we showed that in vitro repetitive stimulation of naïve DO11.10 CD4(+) T cells with Ad-IL-10-DCs resulted in a development of IL-10 producing T-cell regulatory cells. These T cells could not proliferate well but also lost their ability to produce interferon-gamma upon restimulation with irradiated splenocytes and ovalbumin peptide. Furthermore, in co-culture experiments these T cells inhibited the antigen-driven proliferation of naïve CD4+ T cells in a dose-dependent manner. Our findings demonstrated that IL-10 producing DCs had the potential to induce the differentiation of Tr1-like cells and suggested their therapeutic use.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Células Dendríticas/inmunología , Interleucina-10/inmunología , Linfocitos T Reguladores/inmunología , Adenoviridae/genética , Animales , Células de la Médula Ósea/inmunología , Diferenciación Celular , Técnicas de Cocultivo , Femenino , Vectores Genéticos/genética , Vectores Genéticos/farmacología , Tolerancia Inmunológica , Interferón gamma/inmunología , Interleucina-10/genética , Activación de Linfocitos , Ratones , Ratones Endogámicos BALB C , Ratones Transgénicos , Transducción Genética/métodosRESUMEN
Current models of adult haematopoiesis propose that haematopoietic stem cells (HSCs) differentiate into common lymphoid (CLP) and common myeloid (CMP) progenitors and establish an early separation between myeloid and lymphoid lineages. Nevertheless, the developmental potential of CMP-associated B cells suggests the existence of alternate pathways for B lymphopoesis. The aim of this study was to compare the developmental and functional properties of CMP- and CLP-derived B cells. While both populations matured through pro-B cell and transitional B cell intermediates in the bone marrow and spleen, respectively, following transfer into irradiated mice, mature CMP- and CLP-derived B cells exhibit distinct functional responses. Specifically, CMP-derived B cells did not respond to mitogenic stimulation to the same degree as their CLP-derived counterparts and secrete lower levels of IgM and the inflammatory cytokines such as interleukin (IL)-6 and IL-10. Together, these data suggest the existence of multiple pathways for generating functionally distinct B cells from bone marrow precursors.
Asunto(s)
Subgrupos de Linfocitos B/inmunología , Linfopoyesis/inmunología , Células Progenitoras Mieloides/citología , Células Precursoras de Linfocitos B/citología , Traslado Adoptivo , Animales , Antígenos CD19/análisis , Células de la Médula Ósea/inmunología , Diferenciación Celular/inmunología , Células Cultivadas , Citocinas/biosíntesis , Inmunoglobulina M/biosíntesis , Ganglios Linfáticos/inmunología , Ratones , Ratones Endogámicos C57BL , Cavidad Peritoneal/citologíaRESUMEN
The role of the liver in the initiation and maintenance of tolerance is a critical immune function that involves multiple lineages of immune cells. Included within these populations are liver dendritic cells (DCs). Although there has been significant work on the phenotypic and functional roles of splenic and bone marrow dendritic cells, as well as their subsets, comparable studies in liver have often been difficult. To address this issue we have isolated, from C57BL/6 mice, relatively pure populations of DCs and compared phenotype and function to the data from spleen using flow cytometry, cell sorter assisted purification and culture, morphology by cytospin and May-Giemsa staining, cell cycle progression, antigen uptake, cytokine production and allo-activation potential. natural killer (NK)1.1(-)CD11c(+) liver DC subsets (conventional DCs, T cell receptor (TcR)beta(-)NK1.1(-)CD11c(+)B220(-) and plasmacytoid DCs, TcRbeta(-)NK1.1(-)CD11c(+)B220(+)) efficiently endocytose dextran and produce significant levels of tumour necrosis factor (TNF)-alpha, interleukin (IL)-6 and IL-12 p40 in response to Toll-like receptor (TLR) ligands, with responses higher than splenic DCs. There is also a differential capability of hepatic DCs to respond to innate signals. Indeed, CD11c(+) hepatic DCs have a greater capacity to respond to innate stimulation but are less capable of inducing CpG activated-allogeneic T cells. These data suggest that hepatic dendritic cells function as a critical bridge between innate and adaptive immunity and are capable of inducing stronger innate responses with a lower capacity for allo-stimulation than splenic dendritic cells. These properties of liver dendritic cells contribute to their unique role in the induction of tolerance.
Asunto(s)
Antígeno CD11c/análisis , Células Dendríticas/inmunología , Hígado/inmunología , Animales , Antígenos de Superficie/análisis , Ciclo Celular/inmunología , Células Cultivadas , Citocinas/biosíntesis , Endocitosis/inmunología , Femenino , Tolerancia Inmunológica , Inmunofenotipificación , Ligandos , Activación de Linfocitos/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Bazo/inmunología , Linfocitos T/inmunología , Receptores Toll-Like/inmunologíaRESUMEN
Interleukin-1 (IL-1) is a proinflammatory cytokine and IL-1 receptor antagonist (IL-1ra) is a natural inhibitor that binds to IL-1 receptor type I without inducing signal transduction. It is suggested that IL-1 is required for allergen-specific T helper type 2 cell activation and the development of airway hyper-responsiveness (AHR), but the immunologic effect of exogenous IL-1ra in allergic asthma remains unclear. To examine the effect of IL-1ra on airway inflammation and immunoeffector cells in allergic asthma, recombinant adenovirus expressing human IL-1ra (Ad-hIL-1ra) was delivered intranasally into ovalbumin (OVA)-immunized mice. Single intranasal administration of Ad-hIL-1ra before airway antigen challenge in OVA-immunized mice significantly decreased the severity of AHR and reduced pulmonary infiltration of eosinophils and neutrophils. Suppression of IL-5 and eotaxin with concomitant enhancement of interferon gamma in bronchoalveolar lavage fluid was also noted in OVA-immunized mice by administration of Ad-hIL-1ra. In addition, histological studies showed that Ad-hIL-1ra was able to decrease OVA-induced peribronchial inflammation. Taken together, our results indicated that administration of Ad-hIL-1ra may have therapeutic potential for the immunomodulatory treatment of allergic asthma.
Asunto(s)
Adenoviridae/genética , Asma/terapia , Bronquios/inmunología , Terapia Genética/métodos , Vectores Genéticos/administración & dosificación , Proteína Antagonista del Receptor de Interleucina 1/genética , Administración por Inhalación , Animales , Asma/inmunología , Asma/metabolismo , Líquido del Lavado Bronquioalveolar/inmunología , Quimiocina CCL11 , Quimiocinas CC/análisis , Femenino , Ingeniería Genética , Vectores Genéticos/genética , Hipersensibilidad/inmunología , Hipersensibilidad/terapia , Interferón gamma/análisis , Proteína Antagonista del Receptor de Interleucina 1/análisis , Interleucina-5/análisis , Pulmón/inmunología , Ratones , Ratones Endogámicos BALB C , Modelos Animales , Ovalbúmina , Células Th2/inmunología , Transducción Genética/métodosRESUMEN
Recently, sera from children with active Henoch-Schönlein purpura (HSP) have been found to enhance interleukin (IL)-8 production by human umbilical venous endothelial cells (HUVEC). To further determine the possible factor with the ability to enhance endothelial IL-8 production in sera from acute stage of HSP, 10 children with HSP at the acute stage and 10 healthy controls were enrolled. IgA antiendothelial cell antibodies (AECA) were detected by cell-based ELISA. Active sera with or without pretreatment with anti-human IgA antibody, sera of controls, and immunoglobulin A (IgA) derived from sera were used to stimulate the HUVEC. The ability of these factors to enhance endothelial IL-8 production was evaluated. Furthermore, signalling pathways were also assayed by different inhibitors, and confirmed by immunoblotting. Serum levels of IgA AECA in HPS patients at the acute stage were significantly higher than in controls (P < 0.001). The active sera could enhance endothelial IL-8 production (P = 0.004, compared with control sera), and the ability of these sera was mostly abolished when pretreated with fixed anti-human IgA antibody. The supernatant IL-8 levels of endothelial cells stimulated by IgA derived from acute stage of HSP were statistically higher than controls (P < 0.001). PD98059, an inhibitor of ERK phosphorylation, significantly reduced IgA AECA-stimulated endothelial IL-8. IgA AECA also enhanced the phosphorylation of ERK1 with a time-dependent manner. Together with these findings, it is concluded that IgA AECA derived from acute stage of HSP may bind to endothelial and enhance endothelial cells to produce IL-8 via MEK/REK signalling pathway.
Asunto(s)
Autoanticuerpos/inmunología , Vasculitis por IgA/inmunología , Inmunoglobulina A/inmunología , Interleucina-8/inmunología , Enfermedad Aguda , Niño , Células Endoteliales/inmunología , Quinasas MAP Reguladas por Señal Extracelular/antagonistas & inhibidores , Quinasas MAP Reguladas por Señal Extracelular/inmunología , Flavonoides/inmunología , Humanos , Inmunoglobulina A/sangre , Proteína Quinasa 1 Activada por Mitógenos/inmunología , Proteína Quinasa 3 Activada por Mitógenos/inmunología , Fosforilación , Transducción de Señal/inmunología , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
OBJECTIVE: To evaluate the annual incidence and other epidemiological characteristics of Henoch-Schönlein purpura (HSP) among children in Taiwan. METHODS: The records of patients were derived from the research database of the Bureau of National Health Insurance, Taiwan, Republic of China, from January 1999 to December 2002. Children younger than 17 yr of age with the diagnosis of HSP were included into this study. Data for each patient including sex, age, date of onset and length of hospitalization were recorded and analysed. RESULTS: A total of 2759 cases were included with an annual incidence of 12.9 (11.8-13.4) per 100 000 children <17 yr of age. The occurrence of HSP had a peak at the age of 5 to 6 yr. In this study, 1118 (40.5%) patients had been hospitalized at some stage. There were 1454 males and 1305 females, for a male to female ratio of 1.11. Males had a higher annual incidence before the age of 10 yr (P = 0.04), and had a lower incidence than females at older ages (P = 0.02). Disease onset was more common in autumn and winter, and no apparent change in seasonal pattern was noted over 4 yr. CONCLUSIONS: Insurance claim data provide useful information on the epidemiology of HSP in Taiwan. Childhood HSP in Taiwan, with an incidence of 12.9 per 100 000 children, occurs commonly in autumn and winter; and at the age of 5 to 6 yr. The characteristics presented in this study may provide valuable data for understanding and further studies of HSP.
Asunto(s)
Vasculitis por IgA/epidemiología , Adolescente , Distribución por Edad , Niño , Preescolar , Bases de Datos Factuales , Femenino , Encuestas Epidemiológicas , Hospitalización/estadística & datos numéricos , Humanos , Incidencia , Lactante , Recién Nacido , Masculino , Programas Nacionales de Salud/estadística & datos numéricos , Estaciones del Año , Distribución por Sexo , Taiwán/epidemiologíaRESUMEN
Allergic asthma is characterized by airway hyper-responsiveness (AHR) and cellular infiltration of the airway with predominantly eosinophils and Th2 cells. The normal resolution of inflammation in the lung occurs through the regulated removal of unneeded cells by Fas-Fas ligand-mediated apoptosis. Fas ligand (FasL) is a member of the tumor necrosis factor family, and when bound to Fas, it induces apoptosis of the cells. To examine the effect of the FasL gene on airway inflammation and immune effector cells in allergic asthma, recombinant adenovirus expressing murine FasL (Ad-FasL) was delivered intratracheally into ovalbumin (OVA)-immunized mice. We found that a single administration of Ad-FasL in OVA-immunized mice significantly alleviated AHR and eosinophilia by inducing the apoptosis of eosinophils and/or reducing eosinophil attractant factors, such as IL-5 and eotaxin levels. The number of infiltrated lymphocytes and Th2 cytokines, including IL-5 and IL-13, decreased in OVA-immunized mice by administration of Ad-FasL. KC and TNF-alpha production also decreased in Ad-FasL-treated OVA-immunized mice. These findings indicated that the administration of Ad-FasL to OVA-sensitized mice significantly suppressed pulmonary allergic responses. Although more studies are needed, these results suggested that Ad-FasL might be applied as an alternative therapy for allergic asthma.
Asunto(s)
Adenoviridae/genética , Asma/terapia , Terapia Genética/métodos , Vectores Genéticos/administración & dosificación , Glicoproteínas de Membrana/genética , Mucosa Respiratoria/inmunología , Animales , Asma/inmunología , Hiperreactividad Bronquial , Líquido del Lavado Bronquioalveolar/inmunología , Células Dendríticas/inmunología , Proteína Ligando Fas , Femenino , Vectores Genéticos/genética , Recuento de Leucocitos , Ratones , Ratones Endogámicos BALB C , Modelos Animales , Transducción Genética/métodosRESUMEN
BACKGROUND: Pili annulati is a rare autosomal dominant inherited hair shaft abnormality in which clinical examination reveals alternating light and dark bands leading to a shiny appearance of the hair. The clinically light bands are the abnormal areas due to cavities within the cortex. The pathogenesis remains unknown. OBJECTIVES: To investigate the expression of the basement membrane zone (BMZ) components in pili annulati hair follicles of the scalp. METHODS: Transmission electron microscopy (TEM) was carried out on scalp sections of six individuals with pili annulati and six controls. Longitudinal sections of scalp tissues from four individuals with pili annulati and six normal controls were studied by immunohistochemistry with a panel of monoclonal antibodies to the following BMZ components: alpha(6)beta(4) integrin, laminin 5, LH39 antigen, laminin 1, collagen IV and collagen VII. RESULTS: Using TEM, pili annulati scalp specimens exhibited a reduplicated lamina densa in the region of the root bulb in comparison with the single thin electron-dense band in controls. Using immunohistochemistry, there was a wavy BMZ in pili annulati follicles with antibodies to components of the lamina lucida, lamina densa and anchoring fibrils, whereas the BMZ in control hair follicles was as a smooth linear band. The expression of the hemidesmosome-associated alpha(6)beta(4) integrin was linear in both pili annulati and control hair follicles. CONCLUSIONS: Our results suggest that the genetic defect may be a mutation in proteins involved in signalling and regulation of formation and degradation of the lamina densa and sublamina densa region resulting in abnormal assembly or remodelling of the BMZ.
Asunto(s)
Folículo Piloso/anomalías , Adulto , Anticuerpos Monoclonales , Membrana Basal/patología , Femenino , Folículo Piloso/patología , Humanos , Inmunohistoquímica/métodos , Microscopía Electrónica/métodos , Persona de Mediana Edad , Cuero Cabelludo/patologíaRESUMEN
BACKGROUND: The composition of the basement membrane zone (BMZ) or dermal-epidermal junction in the interfollicular skin has been well documented. However, little is known about the BMZ or connective tissue-epithelial junction along the hair follicle. OBJECTIVES: To determine whether the BMZ antigens in the interfollicular epidermis are also present in the BMZ of the anagen hair follicle and to compare whether the expression and distribution of the BMZ components vary between the interfollicular epidermis and the anagen follicle and within different regions of the hair follicle. METHODS: Longitudinal cryostat sections of scalp margin specimens from four adult patients undergoing cosmetic surgery, and without known pathology were stained with a panel of monoclonal and polyclonal antibodies to different BMZ constituents using standard indirect immunofluorescence. RESULTS: All the BMZ antigens found in the normal interfollicular epidermis were expressed in the anagen follicle; however, there were regional variations in the intensity and patterns of fluorescence. All the antigens were expressed in a continuous linear pattern along the BMZ of the interfollicular skin, the infundibulum, and the middle part of the hair follicle. Differences were observed in the lower follicle and the hair bulb. There was continuous expression throughout the BMZ of the follicle of laminin-1 and collagen IV, but in contrast, expression of other antigens decreased down the lower follicle. There was weak or even negative staining with antibodies to alpha 6 beta 4 integrin, laminin-5, anchoring filaments, and type VII collagen in the outer aspect of the bulb compared with the hair papilla. In addition, there were special patterns observed along the bilateral middle and lower follicle. CONCLUSIONS: Despite the common embryological origin between the interfollicular epidermis and the hair follicle, there is variation in the expression of the BMZ antigens. This may be explained by the histological specialization and functional requirements that reflect the dynamic hair growth cycle.
Asunto(s)
Antígenos/metabolismo , Epidermis/metabolismo , Folículo Piloso/metabolismo , Adulto , Membrana Basal/anatomía & histología , Membrana Basal/metabolismo , Tejido Conectivo/anatomía & histología , Tejido Conectivo/metabolismo , Epidermis/anatomía & histología , Técnica del Anticuerpo Fluorescente Indirecta , Folículo Piloso/anatomía & histología , Humanos , Cuero Cabelludo/anatomía & histología , Cuero Cabelludo/metabolismo , Piel/anatomía & histología , Piel/metabolismoRESUMEN
OBJECTIVE: To investigate the roles of cytochrome c and caspases in the pathogenesis of muscular damage in obstructed ureters in a rat model. MATERIALS AND METHODS: Apoptotic cells were detected using in situ end-labelling of DNA fragments. The expression of cytochrome c, and caspases-3, -8 and -9 was examined in 54 rats, using immunohistochemistry. RESULTS: The severity of ureteric smooth muscle damage increased during obstruction. Apoptotic myocytes, and the expression of cytochrome c and the three caspases in the smooth muscle layer were apparent 14 days after ligation, reaching a peak at 21 days. The numbers of apoptotic cells in the smooth muscle layer correlated significantly with expression of cytochrome c and the three caspases (r = 0.8673, 0.8701, 0.5723 and 0.7910, respectively; all P < 0.01). The expression of cytochrome c in the smooth muscle layer correlated significantly with the expression of the three caspases (r = 0.8234, 0.7558 and 0.7825, respectively; all P < 0.001). The expression of caspase-3 and -8, and -3 and -9 also correlated significantly (r = 0.6721 and 0.8501, respectively; both P < 0.002). CONCLUSIONS: Cytochrome c and caspases are involved in ureteric myocyte apoptosis; the release of cytochrome c might be important in ureteric damage during obstructive uropathy.
Asunto(s)
Caspasas/metabolismo , Grupo Citocromo c/metabolismo , Células Musculares/patología , Obstrucción Ureteral/etiología , Animales , Apoptosis , Activación Enzimática , Femenino , Inmunohistoquímica , Etiquetado Corte-Fin in Situ , Modelos Animales , Músculo Liso , Ratas , Ratas Sprague-Dawley , Obstrucción Ureteral/metabolismo , Obstrucción Ureteral/patologíaRESUMEN
Anti-endothelial cell antibodies (AECA) have been found to play an important role in many vascular disorders. In order to determine the presence of AECA in children with Henoch-Schönlein purpura (HSP), and to elucidate the pathogenic and clinical value of their measurement in this disease, AECA were detected by immunofluorescence staining and a human umbilical vein endothelial cell (HUVEC)-based enzyme-linked immunosorbent assay (ELISA) in 20 children with HSP, 10 children with juvenile rheumatoid arthritis (JRA) without vasculitis and 10 normal healthy children. Antibodies against another endothelial cells, human dermal microvascular endothelial cells (HMVEC-d) were also detected by cell-based ELISA. In some experiments, we compared the binding activity of antibodies to HUVEC with and without tumour necrosis factor-alpha (TNF-alpha) or interleukin-1 (IL-1) pretreatment. Patients with acute onset of HSP had higher serum levels of IgA antibodies, both against HUVEC and against HMVEC-d, than healthy controls (P = 0.001, P = 0.008, respectively). Forty-five per cent of patients had positive IgA AECA to HUVEC, and 35% had positive IgA AECA to HMVEC-d. The titres of IgA antibodies to HUVEC paralleled the disease activity. After TNF-alpha treatment, the values of IgA AECA to HUVEC in HSP patients were significantly increased (P = 0.02). For IgG and IgM AECA, there was no difference between HSP patients and controls (P = 0.51, P = 0.91). Ten JRA children without vasculitis had no detectable IgG, IgM or IgA AECA activity. The results of this study showed that children with HSP had IgA AECA, which were enhanced by TNF-alpha treatment. Although the role of these antibodies is not clear, IgA AECA provide another immunological clue for the understanding of HSP.
Asunto(s)
Autoanticuerpos/biosíntesis , Endotelio Vascular/inmunología , Vasculitis por IgA/inmunología , Inmunoglobulina A/biosíntesis , Factor de Necrosis Tumoral alfa/farmacología , Adolescente , Autoanticuerpos/sangre , Cardiolipinas/farmacología , Células Cultivadas , Niño , Preescolar , Citocinas/sangre , Citocinas/farmacología , Dermis/irrigación sanguínea , Endotelio Vascular/efectos de los fármacos , Ensayo de Inmunoadsorción Enzimática , Femenino , Técnica del Anticuerpo Fluorescente , Humanos , Vasculitis por IgA/diagnóstico , Inmunoglobulina A/sangre , Masculino , Venas Umbilicales/citologíaRESUMEN
OBJECTIVE: To determine the role of apoptosis-related proteins (Myc, Bax, Bcl-2, and Bcl-X(L)) in muscular damage in obstructed rat ureters. MATERIALS AND METHODS: The in situ end-labelling of DNA fragments and the expression of apoptosis-related proteins were assessed, using immunohistochemistry, in 54 female Sprague-Dawley rats in which unilateral ureteric obstruction was caused by ureteric ligation. RESULTS: The severity of ureteric damage increased during the period of obstruction. Apoptotic cells and the expression of Bax were detected in the smooth muscle layer from 14 days after ligation. The percentage of apoptotic cells and the expression of Bax in the smooth muscle layer increased and reached a peak 21 days after ligation, and then declined. The expression of Myc was also detected in the smooth muscle layer 14 days after ligation but reached a peak at 28 days. The expressions of Bcl-2 and Bcl-X(L) in the smooth muscle layer were only detected 21 and 28 days after ligation. The numbers of apoptotic cells in the smooth muscle layer correlated significantly with the expressions of Myc and Bax (r = 0.7360 and 0.7432, respectively; both P < 0.005), and with the expression index of Bax/Bcl-2 and Bax/Bcl-X(L) (r = 0.8909 and 0.8592, respectively; both P < 0.001). CONCLUSIONS: Apoptosis-related proteins might be important in regulating cell apoptosis in ureteric damage during the development of obstructive uropathy.
Asunto(s)
Enfermedades Musculares/patología , Proteínas Proto-Oncogénicas c-bcl-2/metabolismo , Proteínas Proto-Oncogénicas c-myc/metabolismo , Proteínas Proto-Oncogénicas/metabolismo , Obstrucción Ureteral/patología , Animales , Apoptosis , Femenino , Inmunohistoquímica/métodos , Etiquetado Corte-Fin in Situ/métodos , Ligadura , Músculo Liso/metabolismo , Músculo Liso/patología , Enfermedades Musculares/metabolismo , Ratas , Ratas Sprague-Dawley , Obstrucción Ureteral/metabolismo , Proteína X Asociada a bcl-2 , Proteína bcl-XRESUMEN
MELAS syndrome (mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes) is one of the classic mitochondrial encephalomyopathies with variable clinical presentation and multisystem involvement. Enhanced sensitivity to neuromuscular blockade or anesthetic agents and susceptibility to malignant hyperthermia in these patients have ever been reported, all of which complicate the management of general anesthesia. To avoid these appalling troubles in general anesthesia, we chose spinal anesthesia for a patient with MELAS syndrome receiving appendectomy. The patient obtained adequate anesthesia and good recovery without neurologic sequelae. Although there is little information about the application of regional anesthesia in MELAS patients, we demonstrate that it may be a satisfactory choice. However, it is suggested that regional anesthesia is performed only when neurological abnormalities of spinal cord or peripheral nerves are definitely ruled out.