RESUMEN
ACE2 is a membrane protein that regulates the cardiovascular system. Additionally, ACE2 acts as a receptor for host cell infection by human coronaviruses, including SARS-CoV-2 that emerged as the cause of the on-going COVID-19 pandemic and has brought unprecedented burden to economy and health. ACE2 binds the spike protein of SARS-CoV-2 with high affinity and shows little variation in amino acid sequence meaning natural resistance is rare. The discovery of a novel short ACE2 isoform (deltaACE2) provides evidence for inter-individual differences in SARS-CoV-2 susceptibility and severity, and likelihood of developing subsequent 'Long COVID'. Critically, deltaACE2 loses SARS-CoV-2 spike protein binding sites in the extracellular domain, and is predicted to confer reduced susceptibility to viral infection. We aimed to assess the differential expression of full-length ACE2 versus deltaACE2 in a panel of human tissues (kidney, heart, lung, and liver) that are implicated in COVID-19, and confirm ACE2 protein in these tissues. Using dual antibody staining, we show that deltaACE2 localises, and is enriched, in lung airway epithelia and bile duct epithelia in the liver. Finally, we also confirm that a fluorescently tagged SARS-CoV-2 spike protein monomer shows low binding at lung and bile duct epithelia where dACE2 is enriched.
Asunto(s)
Enzima Convertidora de Angiotensina 2/metabolismo , SARS-CoV-2/fisiología , Glicoproteína de la Espiga del Coronavirus/metabolismo , Enzima Convertidora de Angiotensina 2/química , Enzima Convertidora de Angiotensina 2/genética , Conductos Biliares/metabolismo , Conductos Biliares/virología , Sitios de Unión , COVID-19/patología , COVID-19/virología , Humanos , Pulmón/metabolismo , Pulmón/virología , Microscopía de Fluorescencia por Excitación Multifotónica , Unión Proteica , Isoformas de Proteínas/química , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Receptores Virales/química , Receptores Virales/metabolismo , SARS-CoV-2/aislamiento & purificación , Glicoproteína de la Espiga del Coronavirus/química , Internalización del VirusRESUMEN
COVID-19 is a pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Early reported symptoms include fever, cough, and respiratory symptoms. There were few reports of digestive symptoms. However, with COVID-19 spreading worldwide, symptoms such as vomiting, diarrhoea, and abdominal pain have gained increasing attention. Research has found that angiotensin-converting enzyme 2 (ACE2), the SARS-CoV-2 receptor, is strongly expressed in the gastrointestinal tract and liver. Whether theoretically or clinically, many studies have suggested a close connection between COVID-19 and the digestive system. In this review, we summarize the digestive symptoms reported in existing research, discuss the impact of SARS-CoV-2 on the gastrointestinal tract and liver, and determine the possible mechanisms and aetiology, such as cytokine storm. In-depth exploration of the relationship between COVID-19 and the digestive system is urgently needed.
Asunto(s)
COVID-19/complicaciones , Enfermedades Gastrointestinales/etiología , Hepatopatías/etiología , Pandemias , SARS-CoV-2/patogenicidad , Enzima Convertidora de Angiotensina 2/metabolismo , Anorexia/etiología , Antivirales/efectos adversos , Conductos Biliares/metabolismo , Conductos Biliares/virología , COVID-19/epidemiología , COVID-19/inmunología , COVID-19/patología , Enfermedad Hepática Inducida por Sustancias y Drogas/etiología , Comorbilidad , Síndrome de Liberación de Citoquinas/etiología , Efecto Citopatogénico Viral , Enfermedades Gastrointestinales/epidemiología , Microbioma Gastrointestinal , Tracto Gastrointestinal/metabolismo , Tracto Gastrointestinal/patología , Tracto Gastrointestinal/virología , Humanos , Inmunosupresores/efectos adversos , Hígado/metabolismo , Hígado/patología , Hígado/virología , Hepatopatías/epidemiología , Trasplante de Hígado , Enfermedad del Hígado Graso no Alcohólico/etiología , Enfermedad del Hígado Graso no Alcohólico/patología , Enfermedad del Hígado Graso no Alcohólico/virología , Complicaciones Posoperatorias , Receptores Virales/metabolismoRESUMEN
The SARS-CoV-2 pandemic has proven to be a serious challenge for the Spanish healthcare system. The impact of the virus on the liver is not well known, but in patients with chronic liver disease, mostly in advanced stages, it can critically compromise survival and trigger decompensation. Treatment in this subpopulation is complex due to the potential hepatotoxicity of some of the medicinal products used. Moreover, the pandemic has also negatively impacted patients with liver disease who have not contracted COVID-19, since the reallocation of human and material resources to the care of patients with the virus has resulted in a decrease in the treatment, diagnosis and follow-up of patients with liver disease, which will surely have negative consequences in the near future. Efficient reorganization of hepatology units is a priority to minimise the impact of the pandemic on a population as vulnerable as liver disease patients.
Asunto(s)
Betacoronavirus , Infecciones por Coronavirus/epidemiología , Hepatopatías/epidemiología , Pandemias , Neumonía Viral/epidemiología , Adenosina Monofosfato/efectos adversos , Adenosina Monofosfato/análogos & derivados , Adenosina Monofosfato/uso terapéutico , Factores de Edad , Alanina/efectos adversos , Alanina/análogos & derivados , Alanina/uso terapéutico , Antivirales/efectos adversos , Antivirales/uso terapéutico , Conductos Biliares/virología , COVID-19 , Enfermedad Hepática Inducida por Sustancias y Drogas/etiología , Enfermedad Crónica , Comorbilidad , Infecciones por Coronavirus/tratamiento farmacológico , Susceptibilidad a Enfermedades , Gastroenterología/organización & administración , Recursos en Salud/provisión & distribución , Hepatitis Crónica/tratamiento farmacológico , Hepatitis Crónica/epidemiología , Humanos , Inmunosupresores/efectos adversos , Hígado/efectos de los fármacos , Hígado/patología , Hígado/virología , Pruebas de Función Hepática , Trasplante de Hígado , Obesidad/epidemiología , Asignación de Recursos , Factores de Riesgo , SARS-CoV-2 , Tratamiento Farmacológico de COVID-19RESUMEN
The review presents the data accumulated for more than 20 years of research of torque teno virus (TTV). Its molecular genetic structure, immunobiology, epidemiology, diagnostic methods, possible replication sites, and pathogenicity factors are described. TTV is a virus that is frequently detectable in patients with different viral hepatitides, in cases of hepatitis without an obvious viral agent, as well as in a healthy population. There is evidence suggesting that biochemical and histological changes occur in liver tissue and bile duct epithelium in TTV monoinfection. There are sufficient histological signs of liver damage, which confirm that the virus can undergo a replicative cycle in hepatocytes. Along with this, cytological hybridization in TTV-infected cells has shown no substantial cytopathic (cell-damaging) effects that are characteristic of pathogenic hepatotropic viruses. Studying TTV has led to the evolution of views on its role in the development of human pathology. The first ideas about the hepatotropism of the virus were gradually reformed as new data became available on the prevalence of the virus and its co-infection with other viruses, including the viruses of the known types of hepatitides. The high prevalence of TTV in the human population indicates its persistence in the body as a virome and a non-pathogenic virus. It has recently been proposed that the level of TTV DNA in the blood of patients undergoing organ transplantation should be used as an endogenous marker of the body's immune status. The available data show the polytropism of the virus and deny the fact that TTV can be assigned exclusively to hepatitis viruses. Fortunately, the rare detection of the damaging effect of TTV on hepatic and bile duct epithelial cells may be indirect evidence of its conditionally pathogenic properties. The ubiquity of the virus and the variability of its existence in humans cannot put an end to its study.
Asunto(s)
Infecciones por Virus ADN/virología , Hepatitis Viral Humana/virología , Hígado/virología , Torque teno virus/patogenicidad , Viroma/inmunología , Conductos Biliares/patología , Conductos Biliares/virología , Infecciones por Virus ADN/sangre , Infecciones por Virus ADN/patología , ADN Viral/aislamiento & purificación , Epitelio/virología , Hepatitis Viral Humana/sangre , Hepatitis Viral Humana/patología , Hepatocitos/virología , Humanos , Hígado/patología , Prevalencia , Torque teno virus/genética , Torque teno virus/inmunología , Carga Viral/inmunologíaRESUMEN
Primary sclerosing cholangitis (PSC) is a chronic, cholestatic liver disease of unknown cause. In the study, we found that duck circovirus (DuCV) induces PSC in natural and reproductive cases. PSC in DuCV naturally infected ducks was investigated by PCR and histopathology. A model of PSC was developed in one-day old duck by infection of DuCV. Effects on serum levels of liver enzymes and histology were evaluated, and DuCV tropism for bile duct in liver was analyzed by immuohistochemistry. Pathology observation of natural or reproductive DuCV infected ducks showed that the lesion of liver were characterized by cholangiocytic injuries and progressive fibrous obliteration of the biliary tree associated with lymphocytes infiltration. ALT, AST, ALP, GGT, ALB, TBIL and TP were significantly increased in serum of DuCV infected ducks. DuCV showed higher tropism for epithelial cells of bile duct than other cells in PSC.
Asunto(s)
Conductos Biliares/virología , Enfermedades de las Aves/fisiopatología , Colangitis Esclerosante/fisiopatología , Colangitis Esclerosante/virología , Circovirus/fisiología , Tropismo Viral/fisiología , Animales , Conductos Biliares/citología , Enfermedades de las Aves/virología , Colangitis Esclerosante/inmunología , Circovirus/inmunología , Patos , Humanos , Hígado/patología , Hígado/virología , Linfocitos/inmunologíaRESUMEN
Cholestatic patients exhibiting high bile acid serum levels were reported to be more susceptible to bacterial and viral infections. Animal studies in bile duct ligated (BDL) mice suggest that cholestasis leads to an aggravation of hepatic bacterial infections. We have investigated the impact of cholestasis on mouse cytomegalovirus (MCMV)-induced immune responses and viral replication. While MCMV did not aggravate BDL-induced liver damage, BDL markedly reduced MCMV-triggered chemokine expression and immune cell recruitment to the liver. MCMV-infected BDL mice showed diminished trafficking of Ly6C+/F4/80+ myeloid cells and NK1.1+ NK cells to the liver compared to MCMV infected control mice. Moreover, virus-driven expression of CCL7, CCL12, CXCL9 and CXCL10 was clearly impaired in BDL- compared to sham-operated mice. Furthermore, production of the anti-inflammatory cytokine IL-10 was massively augmented in infected BDL mice. In contrast, intra- and extrahepatic virus replication was unaltered in BDL-MCMV mice when compared to sham-MCMV mice. Cholestasis in the BDL model severely impaired pathogen-induced chemokine expression in the liver affecting CCR2- and CXCR3-dependent cell trafficking. Cholestasis resulted in reduced recruitment of inflammatory monocytes and NK cells to the liver.
Asunto(s)
Ácidos y Sales Biliares/inmunología , Conductos Biliares/inmunología , Movimiento Celular/inmunología , Colestasis/inmunología , Colestasis/virología , Infecciones por Herpesviridae/inmunología , Células Asesinas Naturales/inmunología , Monocitos/inmunología , Muromegalovirus/fisiología , Replicación Viral/inmunología , Animales , Conductos Biliares/patología , Conductos Biliares/virología , Quimiocinas/inmunología , Colestasis/patología , Infecciones por Herpesviridae/patología , Humanos , Inflamación/inmunología , Inflamación/patología , Inflamación/virología , Células Asesinas Naturales/patología , Ligadura , Masculino , Ratones , Monocitos/patologíaRESUMEN
Biliary atresia (BA) is an infantile inflammatory cholangiopathy of unknown etiology although epidemiologic studies and animal models utilizing rotavirus (RV) have suggested a role for viral infection. Proinflammatory and profibrotic cytokines have been detected in infants with BA. The purpose of our study was to investigate the susceptibility of human cholangiocytes (H69 cells) to infection with RRV and to determine if this infection resulted in cytokine secretion. Infection of H69 cells by RRV was noncytolytic and resulted in a time-dependent increase in the release of both infectious virions and cytokines IL-6 and IL-8 into the supernate. The greatest difference in cytokine supernatant levels between infected and mock-infected cells was noted at 24 hours postinfection (h p.i.) for IL-8, 556 ± 111 versus 77 ± 68 pg/mL (p < 0.0001), and at 48 h p.i. for IL-6, 459 ± 64 versus 67 ± 2 pg/mL (p < 0.0001). Production of both cytokines following RRV infection was significantly reduced by pretreating the H69 cells with inhibitors of mitogen-activated protein kinase (MAPK). Conclusion. RRV can infect human cholangiocytes resulting in the production of proinflammatory and profibrotic cytokines via the MAPK pathway. RRV-infected H69 cells could be a useful model system for investigating the viral hypothesis of BA.
Asunto(s)
Conductos Biliares/inmunología , Conductos Biliares/virología , Interleucina-6/inmunología , Interleucina-8/inmunología , Sistema de Señalización de MAP Quinasas/inmunología , Rotavirus/fisiología , Línea Celular , Humanos , Regulación hacia Arriba/inmunologíaRESUMEN
Biliary atresia (BA), a neonatal obstructive cholangiopathy, remains the most common indication for pediatric liver transplantation in the United States. In the murine model of BA, Rhesus rotavirus (RRV) VP4 surface protein determines biliary duct tropism. In this study, we investigated how VP4 governs induction of murine BA. Newborn mice were injected with 16 strains of rotavirus and observed for clinical symptoms of BA and mortality. Cholangiograms were performed to confirm bile duct obstruction. Livers and bile ducts were harvested 7 days postinfection for virus titers and histology. Flow cytometry assessed mononuclear cell activation in harvested cell populations from the liver. Cytotoxic NK cell activity was determined by the ability of NK cells to kill noninfected cholangiocytes. Of the 16 strains investigated, the 6 with the highest homology to the RRV VP4 (>87%) were capable of infecting bile ducts in vivo. Although the strain Ro1845 replicated to a titer similar to RRV in vivo, it caused no symptoms or mortality. A Ro1845 reassortant containing the RRV VP4 induced all BA symptoms, with a mortality rate of 89%. Flow cytometry revealed that NK cell activation was significantly increased in the disease-inducing strains and these NK cells demonstrated a significantly higher percentage of cytotoxicity against noninfected cholangiocytes. Rotavirus strains with >87% homology to RRV's VP4 were capable of infecting murine bile ducts in vivo. Development of murine BA was mediated by RRV VP4-specific activation of mononuclear cells, independent of viral titers.
Asunto(s)
Atresia Biliar/patología , Proteínas de la Cápside/genética , Colestasis/patología , Leucocitos Mononucleares/fisiología , Activación de Macrófagos/fisiología , Infecciones por Rotavirus/patología , Rotavirus/genética , Animales , Conductos Biliares/virología , Conductos Biliares Extrahepáticos/patología , Interferón gamma/metabolismo , Células Asesinas Naturales/patología , Hígado/patología , Ratones , Ratones Endogámicos BALB C , Filogenia , Infecciones por Rotavirus/mortalidad , Ensayo de Placa Viral , Replicación ViralRESUMEN
Biliary atresia (BA) is a neonatal disease that results in obliteration of the biliary tree. The murine model of BA, which mirrors the human disease, is based upon infection of newborn mice with rhesus rotavirus (RRV), leading to an obstructive cholangiopathy. The purpose of this study was to characterize the temporal relationship between viral infection and the induction of this model. BALB/c mice were infected with RRV on day of life (DOL) 0, 3, 5, and 7. Groups were characterized as early-infection (infection by DOL 3) or late-infection (infection after DOL 5). Early RRV infection induced symptoms in 95% of pups with a mortality rate of 80%. In contrast, late infection caused symptoms in only 50% of mice, and 100% of pups survived. The clinical findings correlated with histological analysis of extrahepatic biliary trees, cytokine expression, and viral titers. Primary murine cholangiocytes isolated, cultured, and infected with RRV yielded higher titers of infectious virus in those harvested from DOL 2 versus DOL 9 mice. Less interferon alpha and beta was produced in DOL 2 versus DOL 9 RRV infected primary cholangiocytes. Injection of BALB/c interferon alpha/beta receptor knockout (IFN-αßR(-/-)) pups at DOL 7 showed increased symptoms (79%) and mortality (46%) when compared to late infected wild type mice. In conclusion, the degree of injury sustained by relatively immature cholangiocytes due to more robust RRV replication correlated with more severe clinical manifestations of cholangiopathy and higher mortality. Interferon alpha production by cholangiocytes appears to play a regulatory role. These findings confirm a temporal dependence of RRV infection in murine BA and begin to define a pathophysiologic role of the maturing cholangiocyte.
Asunto(s)
Conductos Biliares/patología , Conductos Biliares/virología , Atresia Biliar/etiología , Infecciones por Rotavirus/virología , Rotavirus/fisiología , Replicación Viral , Animales , Animales Recién Nacidos , Atresia Biliar/mortalidad , Modelos Animales de Enfermedad , Inflamación/genética , Interferón-alfa/genética , Interferón-alfa/metabolismo , Interferón beta/genética , Interferón beta/metabolismo , Hígado/enzimología , Ratones , Receptor de Interferón alfa y beta/deficiencia , Receptor de Interferón alfa y beta/genética , Infecciones por Rotavirus/metabolismo , Factores de TiempoRESUMEN
Structural analysis of hepatocytes in liver biopsy specimens from patients with hepatitis C and C+B with intracellular cholestasis was carried out. Large foci of bile components in hepatocytes led to cell damage, eventuating in cell destruction and death. The cholestatic variant of mixed infection was characterized by destructive necrotic changes in hepatocytes and progressive fibrosis of the liver. Destruction of the hepatocyte cytoplasmic organelles was associated with high activity of the infectious process and pronounced cytolytic syndrome.
Asunto(s)
Conductos Biliares/patología , Colestasis/patología , Hepacivirus , Virus de la Hepatitis B , Hepatitis B Crónica/patología , Hepatitis C Crónica/patología , Hepatocitos/patología , Hígado/patología , Conductos Biliares/virología , Biopsia , Muerte Celular , Colestasis/complicaciones , Colestasis/virología , Coinfección , Femenino , Fibrosis , Hepatitis B Crónica/complicaciones , Hepatitis B Crónica/virología , Hepatitis C Crónica/complicaciones , Hepatitis C Crónica/virología , Hepatocitos/virología , Humanos , Hígado/virología , Masculino , Índice de Severidad de la EnfermedadRESUMEN
PURPOSE OF REVIEW: New knowledge on rotavirus infection in children and well established mouse models has renewed interest in whether rotavirus could cause biliary atresia, an idiopathic, obliterative infantile disease of bile ducts that is the primary indication for liver transplant in children. RECENT FINDINGS: Studies in the rotavirus mouse model of biliary atresia indicate that infection of biliary epithelium is an inaugural event leading to biliary inflammation and obstruction, which is preceded by systemic spread of rotavirus, which also occurs during human rotavirus enteric infections. Viral factors, including rotavirus gene 4, are important for biliary infection and biliary atresia in mice. Specific host factors related to inflammatory processes (natural killer and T cells, interferon) are also critical, and a paucity of regulatory T cells in neonates may play a key role in pathogenesis in experimental biliary atresia. Rotavirus vaccination has substantially decreased rotavirus diarrheal disease worldwide and might enable demonstration of a cause-effect relationship between rotavirus infection and biliary atresia in humans. SUMMARY: Rotavirus can be detected in the serum of mice and children and causes biliary atresia in neonatal mice. Approaches to re-examine whether rotavirus causes biliary atresia in children are discussed based on concepts from the mouse model of biliary atresia and rotavirus vaccination programs.
Asunto(s)
Conductos Biliares/virología , Atresia Biliar , Infecciones por Rotavirus , Rotavirus/aislamiento & purificación , Animales , Atresia Biliar/epidemiología , Atresia Biliar/etiología , Atresia Biliar/virología , Salud Global , Humanos , Incidencia , Lactante , Factores de Riesgo , Infecciones por Rotavirus/complicaciones , Infecciones por Rotavirus/epidemiología , Infecciones por Rotavirus/virologíaRESUMEN
Notwithstanding numerous evidences implicating toll-like receptor-4 (TLR4) in the pathogenesis of chronic hepatitis C virus (HCV) infection, the localization and level of TLR4 expression in the liver of patients with hepatitis C have never been investigated. We aimed to evaluate, by means of immunohistochemistry and real-time PCR (rt-PCR), hepatic TLR4 expression in patients with chronic HCV infection. Fifty patients who had undergone liver biopsy and 11 patients transplanted because of chronic HCV infection, and 12 controls free of liver disease, were included in the study. Each case was analyzed by immunohistochemistry for TLR4, α-smooth muscle actin and cytokeratin-7 (CK-7), and a subgroup of patients and all controls by rt-PCR for TLR4. Immunohistochemistry for α-smooth muscle actin was used to derive a score of activation of hepatic stellate cells and portal/septal myofibroblasts, while immunohistochemistry for CK-7 was used to evaluate and count hepatic progenitor cells, interlobular bile ducts and intermediate hepatocytes. In patients, the parenchymal elements responsible for the highest TLR4 level of expression were hepatic progenitor cells and biliary epithelial cells of interlobular bile ducts. Double-labeling experiments between anti-TLR4 and anti-CK7, anti-CD133, anti-CD44, anti-neural cell adhesion molecule, anti-epithelial cell adhesion molecule and anti-sex determining region Y-box 9, confirmed these findings. TLR4-positive hepatic progenitor cells and interlobular bile ducts were significantly correlated with the stage of liver disease (P<0.001), the grade of inflammation (P<0.001), and the activity of portal/septal myofibroblasts (P<0.001). rt-PCR study confirmed an increased TLR4 expression in the 26 patients analyzed with respect to controls (P<0.001). TLR4 expression positively correlated with fibrosis (P<0.05) and inflammation (P<0.05). The present results suggest that TLR4 expression by hepatic progenitor cells and biliary epithelial cells contributes to the progression of liver damage in the course of chronic HCV-related infection.
Asunto(s)
Conductos Biliares/inmunología , Células Epiteliales/inmunología , Hepatitis C Crónica/inmunología , Cirrosis Hepática/inmunología , Hígado/inmunología , Células Madre/inmunología , Receptor Toll-Like 4/análisis , Adulto , Anciano , Análisis de Varianza , Conductos Biliares/patología , Conductos Biliares/virología , Biomarcadores/análisis , Biopsia , Estudios de Casos y Controles , Distribución de Chi-Cuadrado , Progresión de la Enfermedad , Células Epiteliales/patología , Células Epiteliales/virología , Femenino , Células Estrelladas Hepáticas/inmunología , Células Estrelladas Hepáticas/patología , Células Estrelladas Hepáticas/virología , Hepatitis C Crónica/genética , Hepatitis C Crónica/patología , Hepatocitos/inmunología , Hepatocitos/patología , Hepatocitos/virología , Humanos , Inmunohistoquímica , Hígado/patología , Hígado/virología , Cirrosis Hepática/genética , Cirrosis Hepática/patología , Cirrosis Hepática/virología , Masculino , Microscopía Fluorescente , Persona de Mediana Edad , Miofibroblastos/inmunología , Miofibroblastos/patología , Miofibroblastos/virología , ARN Mensajero/análisis , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Ciudad de Roma , Células Madre/patología , Células Madre/virología , Receptor Toll-Like 4/genéticaRESUMEN
Biliary atresia is a common disease in neonates which causes obstructive jaundice and progressive hepatic fibrosis. Our previous studies indicate that rotavirus infection is an initiator in the pathogenesis of experimental biliary atresia (BA) through the induction of increased nuclear factor-kappaB and abnormal activation of the osteopontin inflammation pathway. In the setting of rotavirus infection, rotavirus nonstructural protein 4 (NSP4) serves as an important immunogen, viral protein 7 (VP7) is necessary in rotavirus maturity and viral protein 4 (VP4) is a virulence determiner. The purpose of the current study is to clarify the roles of NSP4, VP7 and VP4 in the pathogenesis of experimental BA. Primary cultured extrahepatic biliary epithelia were infected with Rotavirus (mmu18006). Small interfering RNA targeting NSP4, VP7 or VP4 was transfected before rotavirus infection both in vitro and in vivo. We analyzed the incidence of BA, morphological change, morphogenesis of viral particles and viral mRNA and protein expression. The in vitro experiments showed NSP4 silencing decreased the levels of VP7 and VP4, reduced viral particles and decreased cytopathic effect. NSP4-positive cells had strongly positive expression of integrin subunit α2. Silencing of VP7 or VP4 partially decreased epithelial injury. Animal experiments indicated after NSP4 silencing, mouse pups had lower incidence of BA than after VP7 or VP4 silencing. However, 33.3% of VP4-silenced pups (Nâ=â6) suffered BA and 50% of pups (Nâ=â6) suffered biliary injury after VP7 silencing. Hepatic injury was decreased after NSP4 or VP4 silencing. Neither VP4 nor VP7 were detected in the biliary ducts after NSP4. All together, NSP4 silencing down-regulates VP7 and VP4, resulting in decreased incidence of BA.
Asunto(s)
Atresia Biliar/patología , Atresia Biliar/virología , Silenciador del Gen , Glicoproteínas/genética , Rotavirus/metabolismo , Toxinas Biológicas/genética , Proteínas no Estructurales Virales/genética , Animales , Conductos Biliares/patología , Conductos Biliares/ultraestructura , Conductos Biliares/virología , Línea Celular , Efecto Citopatogénico Viral , Modelos Animales de Enfermedad , Epitelio/patología , Epitelio/ultraestructura , Epitelio/virología , Técnica del Anticuerpo Fluorescente , Incidencia , Integrina alfa2/metabolismo , Ratones , ARN Interferente Pequeño/metabolismo , Rotavirus/fisiología , Rotavirus/ultraestructura , Replicación ViralRESUMEN
Biliary atresia is the most common cholangiopathy of childhood. With complete obstruction of segments or the entire length of extrahepatic bile ducts, the timely pursuit of hepatoportoenterostomy is the best strategy to restore bile drainage. However, even with prompt surgical intervention, ongoing injury of intrahepatic bile ducts and progressive cholangiopathy lead to end-stage cirrhosis. The pace of disease progression is not uniform; it may relate to clinical forms of disease and/or staging of liver pathology at diagnosis. Although the etiology of disease is not yet defined, several biological processes have been linked to pathogenic mechanisms of bile duct injury. Among them, there is increasing evidence that the immune system targets the duct epithelium and disrupts bile flow. We discuss how careful clinical phenotyping, staging of disease, and basic mechanistic research are providing insights into clinical trial designs and directions for development of new therapies to block progression of disease.
Asunto(s)
Antiinflamatorios/uso terapéutico , Atresia Biliar/tratamiento farmacológico , Inflamación/tratamiento farmacológico , Animales , Conductos Biliares/crecimiento & desarrollo , Conductos Biliares/inmunología , Conductos Biliares/virología , Atresia Biliar/etiología , Atresia Biliar/patología , Ensayos Clínicos como Asunto , Femenino , Humanos , Inflamación/inmunología , Inflamación/virología , Hígado/crecimiento & desarrollo , Hígado/inmunología , Hígado/virología , Cirrosis Hepática/etiología , Cirrosis Hepática/inmunología , Masculino , Ratones , Portoenterostomía Hepática , Resultado del TratamientoRESUMEN
Although PCR-based in situ hybridization (PCR-ISH) can be used to determine the distribution and localization of pathogens in tissues, this approach is hampered by its low specificity. Therefore, we used a highly specific and sensitive PCR-ISH method to reveal the lobular distribution and intracellular localization of hepatitis B virus (HBV) and HCV in chronic liver disease and to clarify the state of persistent HBV and HCV infection in the liver. HBV genomic DNA was detected in almost all hepatocytes, whereas HBV RNA or protein was differentially distributed only in a subset of the HBV DNA-positive region. Further, HCV genomic RNA was detected in almost all hepatocytes and was localized to the cytoplasm. HCV RNA was also detected in the epithelium of the large bile duct but not in endothelial cells, portal tracts, or sinusoidal lymphocytes. In patients with HBV and HCV coinfection, HCV RNA was localized to the noncancerous tissue, whereas HBV DNA was found only in the cancerous tissue. Using this novel PCR-ISH method, we could visualize the staining pattern of HBV and HCV in liver sections, and we obtained results consistent with those of real-time detection (RTD)-PCR analysis. In conclusion, almost all hepatocytes are infected with HBV or HCV in chronic liver disease; this finding implies that the viruses spreads throughout the liver in the chronic stage.
Asunto(s)
Hepacivirus/aislamiento & purificación , Virus de la Hepatitis B/aislamiento & purificación , Hepatitis B Crónica/virología , Hepatitis C Crónica/virología , Hibridación in Situ/métodos , Hígado/virología , Reacción en Cadena de la Polimerasa/métodos , Adulto , Anciano , Conductos Biliares/virología , Femenino , Hepacivirus/genética , Virus de la Hepatitis B/genética , Hepatitis B Crónica/patología , Hepatitis C Crónica/patología , Hepatocitos/virología , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Cellular senescence is defined as irreversible cell arrest and could work as a safeguard against tumorigenesis. This mechanism was examined in chronic viral hepatitis-related hepatocarcinogenesis. By using surgical resected or wedge biopsied liver specimens from 87 chronic viral hepatitis patients in whom 35 neoplastic nodules (dysplastic nodules and hepatocellular carcinoma) were complicated, P21 expression and senescence-associated beta galactosidase activity, a marker of senescence, were examined. All of these neoplastic nodules harbored portal tracts within the tumors. Hepatocytes expressing senescence markers and cytokeratin (CK)7-positive bile ductules including hepatic progenitor-like cells were increased in periseptal areas in cirrhosis. Interestingly, these cells appeared to form an anatomical complex that was completely lost in the periportal areas within the neoplastic nodules. In one-third of the neoplastic nodules, CK7-positive small neoplastic hepatocytes resembling hepatic progenitor cells proliferated zonally around the portal tracts. In conclusion, loss of a complex of senescent hepatocytes and ductular cell including hepatic progenitor-like cells in the periportal or periseptal areas may be associated with emergence of neoplastic hepatocytes and their proliferation followed by neoplastic nodules arising in liver cirrhosis. Zonal proliferation of CK7-positive small neoplastic hepatocytes resembling hepatic progenitor cells may develop during early hepatocarcinogenesis.
Asunto(s)
Conductos Biliares/patología , Transformación Celular Neoplásica/patología , Senescencia Celular/fisiología , Hepatitis Crónica/patología , Hepatocitos/patología , Neoplasias Hepáticas/patología , Anciano , Conductos Biliares/virología , Femenino , Hepatitis B Crónica , Hepatitis C Crónica , Hepatitis Crónica/virología , Hepatocitos/virología , Humanos , Inmunohistoquímica , Neoplasias Hepáticas/virología , Masculino , Persona de Mediana Edad , Células Madre/patologíaRESUMEN
Biliary atresia is a devastating disorder of the newborn in which afflicted infants develop inflammation and fibrosis of the extrahepatic biliary tract, resulting in cirrhosis and end-stage liver disease. Infection with a virus is thought to be a contributing factor in the etiology of biliary atresia. In the murine model of biliary atresia, perinatal exposure to rhesus rotavirus (RRV) results in biliary epithelial cell infection causing bile duct obstruction. The purpose of this study was to determine if tropism for the biliary epithelial cell was unique to RRV. Newborn mice underwent intraperitoneal injection with five strains of rotavirus: RRV (simian), SA11-FM (simian/bovine), SA11-SM (simian), EDIM (murine), and Wa (human). RRV and SA11-FM caused clinical manifestations of bile duct obstruction and high mortality. SA11-SM caused clinical signs of hepatobiliary injury but the mortality was markedly reduced. EDIM and Wa caused no sign of hepatobiliary disease. The systemic and temporal distribution of viral protein and live virus varied according to the injected strain. Immunohistochemistry revealed that RRV and SA11-FM targeted the biliary epithelial cells. In contrast, SA11-SM was found in the liver but in not in the biliary epithelium. These results indicate that strain-specific characteristics dictate tropism for cells of hepatobiliary origin which in turn impact the ability to induce the murine model of biliary atresia.
Asunto(s)
Atresia Biliar/virología , Infecciones por Rotavirus/virología , Rotavirus/patogenicidad , Animales , Animales Recién Nacidos , Conductos Biliares/patología , Conductos Biliares/virología , Atresia Biliar/fisiopatología , Línea Celular , Colestasis/patología , Células Epiteliales/virología , Femenino , Inmunohistoquímica , Hígado/virología , Ratones , Ratones Endogámicos BALB C , Rotavirus/aislamiento & purificación , Especificidad de la Especie , Proteínas Virales/metabolismoRESUMEN
Lentiviral vectors can stably transduce dividing and nondividing cells in vivo and are best suited to long-term correction of inherited liver diseases. Intraportal administration of lentiviral vectors expressing green fluorescent protein (Lenti-GFP) in mice resulted in a higher transduction of nonparenchymal cells than hepatocytes (7.32 +/- 3.66% vs 0.22 +/- 0.08%, respectively). Therefore, various treatments were explored to increase lentiviral transduction of hepatocytes. Lenti-GFP was injected into the common bile duct, which led to transduction of biliary epithelium and hepatocytes at low efficiency. Transient removal of the sinusoidal endothelial cell layer by cyclophosphamide to increase accessibility to hepatocytes did not improve hepatocyte transduction (0.42 +/- 0.36%). Inhibition of Kupffer cell function by gadolinium chloride led to a significant decrease in GFP-positive nonparenchymal cells (2.15 +/- 3.14%) and a sevenfold increase in GFP-positive hepatocytes compared to nonpretreated mice (1.48 +/- 2.01%). These findings suggest that sinusoidal endothelial cells do not significantly limit lentiviral transduction of hepatocytes, while Kupffer cells sequester lentiviral particles thereby preventing hepatocyte transduction. Therefore, the use of agents that inhibit Kupffer cell function may be important for lentiviral vector treatment of liver disease.
Asunto(s)
Hepatocitos/virología , Macrófagos del Hígado/fisiología , Lentivirus/genética , Lentivirus/fisiología , Transducción Genética/métodos , Alanina Transaminasa/metabolismo , Animales , Conductos Biliares/virología , Línea Celular , Ciclofosfamida/farmacología , Células Endoteliales/patología , Células Endoteliales/virología , Gadolinio/farmacología , Hepatocitos/enzimología , Hepatocitos/metabolismo , Macrófagos del Hígado/patología , Lentivirus/efectos de los fármacos , Hígado/enzimología , Hígado/metabolismo , Hígado/ultraestructura , Hígado/virología , Masculino , Ratones , Microscopía Electrónica , Fagocitosis , Reacción en Cadena de la Polimerasa , Vena Porta/virologíaRESUMEN
Canine distemper virus (CDV) caused epizootics in lions (Panthera leo) in Tanzania's Serengeti National Park in 1994 and in captive lions and other Panthera spp. in the USA in 1991-1992. In this study, immunohistochemistry was used to compare viral distribution in tissues collected from ferrets (Mustela putorius furo) inoculated with one of the two lion-derived CDV isolates, either from Serengeti (A94-11/13) or from California (A92-27/20). The California isolate resulted in severe morbidity in all nine ferrets, whereas the Serengeti isolate resulted in severe morbidity in five of the nine ferrets. A slightly higher proportion of infected cells was found in many tissues in the Serengeti isolate-inoculated ferrets. These findings indicate that the pathogenicity of the California isolate is not directly related to the number of infected cells.
Asunto(s)
Virus del Moquillo Canino/clasificación , Virus del Moquillo Canino/aislamiento & purificación , Hurones/virología , Leones/virología , Animales , Conductos Biliares/virología , Encéfalo/virología , Moquillo/virología , Epitelio/virología , Inmunohistoquímica , Tejido Linfoide/virología , Sistema Respiratorio/virología , Glándulas Salivales/virología , Piel/virologíaRESUMEN
Infection of neonatal mice with some reovirus strains produces a disease similar to infantile biliary atresia, but previous attempts to correlate reovirus infection with this disease have yielded conflicting results. We used isogenic reovirus strains T3SA- and T3SA+, which differ solely in the capacity to bind sialic acid as a coreceptor, to define the role of sialic acid in reovirus encephalitis and biliary tract infection in mice. Growth in the intestine was equivalent for both strains following peroral inoculation. However, T3SA+ spread more rapidly from the intestine to distant sites and replicated to higher titers in spleen, liver, and brain. Strikingly, mice infected with T3SA+ but not T3SA- developed steatorrhea and bilirubinemia. Liver tissue from mice infected with T3SA+ demonstrated intense inflammation focused at intrahepatic bile ducts, pathology analogous to that found in biliary atresia in humans, and high levels of T3SA+ antigen in bile duct epithelial cells. T3SA+ bound 100-fold more efficiently than T3SA- to human cholangiocarcinoma cells. These observations suggest that the carbohydrate-binding specificity of a virus can dramatically alter disease in the host and highlight the need for epidemiologic studies focusing on infection by sialic acid-binding reovirus strains as a possible contributor to the pathogenesis of neonatal biliary atresia.