RESUMEN
The liver has a unique ability to regenerate1,2; however, in the setting of acute liver failure (ALF), this regenerative capacity is often overwhelmed, leaving emergency liver transplantation as the only curative option3-5. Here, to advance understanding of human liver regeneration, we use paired single-nucleus RNA sequencing combined with spatial profiling of healthy and ALF explant human livers to generate a single-cell, pan-lineage atlas of human liver regeneration. We uncover a novel ANXA2+ migratory hepatocyte subpopulation, which emerges during human liver regeneration, and a corollary subpopulation in a mouse model of acetaminophen (APAP)-induced liver regeneration. Interrogation of necrotic wound closure and hepatocyte proliferation across multiple timepoints following APAP-induced liver injury in mice demonstrates that wound closure precedes hepatocyte proliferation. Four-dimensional intravital imaging of APAP-induced mouse liver injury identifies motile hepatocytes at the edge of the necrotic area, enabling collective migration of the hepatocyte sheet to effect wound closure. Depletion of hepatocyte ANXA2 reduces hepatocyte growth factor-induced human and mouse hepatocyte migration in vitro, and abrogates necrotic wound closure following APAP-induced mouse liver injury. Together, our work dissects unanticipated aspects of liver regeneration, demonstrating an uncoupling of wound closure and hepatocyte proliferation and uncovering a novel migratory hepatocyte subpopulation that mediates wound closure following liver injury. Therapies designed to promote rapid reconstitution of normal hepatic microarchitecture and reparation of the gut-liver barrier may advance new areas of therapeutic discovery in regenerative medicine.
Asunto(s)
Fallo Hepático Agudo , Regeneración Hepática , Animales , Femenino , Humanos , Masculino , Ratones , Acetaminofén/farmacología , Linaje de la Célula , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Modelos Animales de Enfermedad , Factor de Crecimiento de Hepatocito/metabolismo , Factor de Crecimiento de Hepatocito/farmacología , Hepatocitos/citología , Hepatocitos/efectos de los fármacos , Hepatocitos/metabolismo , Hepatocitos/patología , Hígado/citología , Hígado/efectos de los fármacos , Hígado/patología , Fallo Hepático Agudo/patología , Fallo Hepático Agudo/inducido químicamente , Regeneración Hepática/efectos de los fármacos , Ratones Endogámicos C57BL , Necrosis/inducido químicamente , Medicina Regenerativa , Análisis de Expresión Génica de una Sola Célula , Cicatrización de HeridasRESUMEN
Liver cirrhosis is a major cause of death worldwide and is characterized by extensive fibrosis. There are currently no effective antifibrotic therapies available. To obtain a better understanding of the cellular and molecular mechanisms involved in disease pathogenesis and enable the discovery of therapeutic targets, here we profile the transcriptomes of more than 100,000 single human cells, yielding molecular definitions for non-parenchymal cell types that are found in healthy and cirrhotic human liver. We identify a scar-associated TREM2+CD9+ subpopulation of macrophages, which expands in liver fibrosis, differentiates from circulating monocytes and is pro-fibrogenic. We also define ACKR1+ and PLVAP+ endothelial cells that expand in cirrhosis, are topographically restricted to the fibrotic niche and enhance the transmigration of leucocytes. Multi-lineage modelling of ligand and receptor interactions between the scar-associated macrophages, endothelial cells and PDGFRα+ collagen-producing mesenchymal cells reveals intra-scar activity of several pro-fibrogenic pathways including TNFRSF12A, PDGFR and NOTCH signalling. Our work dissects unanticipated aspects of the cellular and molecular basis of human organ fibrosis at a single-cell level, and provides a conceptual framework for the discovery of rational therapeutic targets in liver cirrhosis.
Asunto(s)
Células Endoteliales/patología , Cirrosis Hepática/patología , Hígado/patología , Macrófagos/patología , Análisis de la Célula Individual , Animales , Estudios de Casos y Controles , Linaje de la Célula , Sistema del Grupo Sanguíneo Duffy/metabolismo , Células Endoteliales/metabolismo , Femenino , Células Estrelladas Hepáticas/citología , Células Estrelladas Hepáticas/metabolismo , Células Estrelladas Hepáticas/patología , Hepatocitos/citología , Hepatocitos/metabolismo , Hepatocitos/patología , Humanos , Hígado/citología , Cirrosis Hepática/genética , Macrófagos/metabolismo , Masculino , Glicoproteínas de Membrana/metabolismo , Proteínas de la Membrana/metabolismo , Ratones , Fenotipo , Receptor alfa de Factor de Crecimiento Derivado de Plaquetas/metabolismo , Receptores de Superficie Celular/metabolismo , Receptores Inmunológicos/metabolismo , Tetraspanina 29/metabolismo , Transcriptoma , Migración Transendotelial y TransepitelialRESUMEN
BACKGROUND: Gallbladder cancer is rare, but cancers detected incidentally after cholecystectomy are increasing. The aim of this study was to review the available data for current best practice for optimal management of incidental gallbladder cancer. METHODS: A systematic PubMed search of the English literature to May 2018 was conducted. RESULTS: The search identified 12 systematic reviews and meta-analyses, in addition to several consensus reports, multi-institutional series and national audits. Some 0·25-0·89 per cent of all cholecystectomy specimens had incidental gallbladder cancer on pathological examination. Most patients were staged with pT2 (about half) or pT1 (about one-third) cancers. Patients with cancers confined to the mucosa (T1a or less) had 5-year survival rates of up to 100 per cent after cholecystectomy alone. For cancers invading the muscle layer of the gallbladder wall (T1b or above), reresection is recommended. The type, extent and timing of reresection remain controversial. Observation time may be used for new cross-sectional imaging with CT and MRI. Perforation at initial surgery had a higher risk of disease dissemination. Gallbladder cancers are PET-avid, and PET may detect residual disease and thus prevent unnecessary surgery. Routine laparoscopic staging before reresection is not warranted for all stages. Risk of peritoneal carcinomatosis increases with each T category. The incidence of port-site metastases is about 10 per cent. Routine resection of port sites has no effect on survival. Adjuvant chemotherapy is poorly documented and probably underused. CONCLUSION: Management of incidental gallbladder cancer continues to evolve, with more refined suggestions for subgroups at risk and a selective approach to reresection.
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Colecistectomía , Neoplasias de la Vesícula Biliar/patología , Neoplasias de la Vesícula Biliar/terapia , Complicaciones Posoperatorias/terapia , Biomarcadores de Tumor/metabolismo , Quimioterapia Adyuvante/estadística & datos numéricos , Conversión a Cirugía Abierta/estadística & datos numéricos , Humanos , Hallazgos Incidentales , Laparoscopía/efectos adversos , Laparoscopía/estadística & datos numéricos , Metástasis de la Neoplasia , Siembra Neoplásica , Complicaciones Posoperatorias/patología , Pronóstico , Reoperación/estadística & datos numéricos , Medición de RiesgoRESUMEN
The number of patients diagnosed with chronic bile duct disease is increasing and in most cases these diseases result in chronic ductular scarring, necessitating liver transplantation. The formation of ductular scaring affects liver function; however, scar-generating portal fibroblasts also provide important instructive signals to promote the proliferation and differentiation of biliary epithelial cells. Therefore, understanding whether we can reduce scar formation while maintaining a pro-regenerative microenvironment will be essential in developing treatments for biliary disease. Here, we describe how regenerating biliary epithelial cells express Wnt-Planar Cell Polarity signalling components following bile duct injury and promote the formation of ductular scars by upregulating pro-fibrogenic cytokines and positively regulating collagen-deposition. Inhibiting the production of Wnt-ligands reduces the amount of scar formed around the bile duct, without reducing the development of the pro-regenerative microenvironment required for ductular regeneration, demonstrating that scarring and regeneration can be uncoupled in adult biliary disease and regeneration.
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Enfermedades de los Conductos Biliares/patología , Colangitis Esclerosante/patología , Cicatriz/patología , Vía de Señalización Wnt , Animales , Proteína Axina/genética , Proteína Axina/metabolismo , Enfermedades de los Conductos Biliares/inducido químicamente , Enfermedades de los Conductos Biliares/metabolismo , Conductos Biliares/citología , Polaridad Celular , Colangitis Esclerosante/metabolismo , Cicatriz/metabolismo , Modelos Animales de Enfermedad , Células Epiteliales , Humanos , Proteínas Quinasas JNK Activadas por Mitógenos/metabolismo , MAP Quinasa Quinasa 4/metabolismo , Masculino , Ratones Transgénicos , Proteínas del Tejido Nervioso/genética , Proteínas del Tejido Nervioso/metabolismo , Piridinas/toxicidad , Vía de Señalización Wnt/efectos de los fármacos , Proteína Wnt-5a/metabolismoRESUMEN
BACKGROUND: Validated diagnostic tools that are accurate, cost effective and acceptable to patients are required for disease stratification and monitoring in NAFLD. AIMS: To investigate the performance and cost of multiparametric MRI alongside existing biomarkers in the assessment of NAFLD. METHODS: Adult patients undergoing standard of care liver biopsy for NAFLD were prospectively recruited at two UK liver centres and underwent multiparametric MRI, blood sampling and transient elastography withing 2 weeks of liver biopsy. Non-invasive markers were compared to histology as the gold standard. RESULTS: Data were obtained in 50 patients and 6 healthy volunteers. Corrected T1 (cT1) correlated with NAFLD activity score (ρ = 0.514, P < .001). cT1, enhanced liver fibrosis (ELF) test and liver stiffness differentiated patients with simple steatosis and NASH with AUROC (95% CI) of 0.69 (0.50-0.88), 0.87 (0.77-0.79) and 0.82 (0.70-0.94) respectively and healthy volunteers from patients with AUROC (95% CI) of 0.93 (0.86-1.00), 0.81 (0.69-0.92) and 0.89 (0.77-1.00) respectively. For the risk stratification of NAFLD, multiparametric MRI could save £150,218 per 1000 patients compared to biopsy. Multiparametric MRI did not discriminate between individual histological fibrosis stages in this population (P = .068). CONCLUSIONS: Multiparametric MRI accurately identified patients with steatosis, stratifies those with NASH or simple steatosis and reliably excludes clinically significant liver disease with superior negative predictive value (83.3%) to liver stiffness (42.9%) and ELF (57.1%). For the risk stratification of NAFLD, multiparametric MRI was cost effective and, combined with transient elastography, had the lowest cost per correct diagnosis.
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Hígado/diagnóstico por imagen , Imagen por Resonancia Magnética , Enfermedad del Hígado Graso no Alcohólico/diagnóstico , Adolescente , Adulto , Anciano , Biopsia , Análisis Costo-Beneficio , Diagnóstico por Imagen de Elasticidad/economía , Diagnóstico por Imagen de Elasticidad/métodos , Femenino , Voluntarios Sanos , Humanos , Hígado/patología , Cirrosis Hepática/diagnóstico , Cirrosis Hepática/economía , Imagen por Resonancia Magnética/economía , Imagen por Resonancia Magnética/métodos , Masculino , Persona de Mediana Edad , Enfermedad del Hígado Graso no Alcohólico/economía , Enfermedad del Hígado Graso no Alcohólico/patología , Valor Predictivo de las Pruebas , Adulto JovenRESUMEN
Mesenchymal cells expressing platelet-derived growth factor receptor beta (PDGFRß) are known to be important in fibrosis of organs such as the liver and kidney. Here we show that PDGFRß+ cells contribute to skeletal muscle and cardiac fibrosis via a mechanism that depends on αv integrins. Mice in which αv integrin is depleted in PDGFRß+ cells are protected from cardiotoxin and laceration-induced skeletal muscle fibrosis and angiotensin II-induced cardiac fibrosis. In addition, a small-molecule inhibitor of αv integrins attenuates fibrosis, even when pre-established, in both skeletal and cardiac muscle, and improves skeletal muscle function. αv integrin blockade also reduces TGFß activation in primary human skeletal muscle and cardiac PDGFRß+ cells, suggesting that αv integrin inhibitors may be effective for the treatment and prevention of a broad range of muscle fibroses.
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Integrina alfaV/metabolismo , Músculo Esquelético/patología , Miocardio/patología , Receptor beta de Factor de Crecimiento Derivado de Plaquetas/genética , Animales , Apoptosis , Movimiento Celular , Células Cultivadas , Colágeno/metabolismo , Fibrosis , Genotipo , Humanos , Masculino , Células Madre Mesenquimatosas/citología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Receptor beta de Factor de Crecimiento Derivado de Plaquetas/metabolismo , Proteínas Recombinantes/metabolismoRESUMEN
Adult testicular dermoid tumours are rare tumours with no reported potential for recurrent or metastatic spread. Despite this they are currently classified as mature teratoma and managed as if they have equivalent malignant potential. This report describes two cases of adult mature teratoma of dermoid type and questions the classification and pathogenesis of this disease. In one of the cases there was a clear history of a testicular lump arising pre-pubertally, raising the possibility that some adult dermoid tumours may in fact be pre-pubertal teratomas that have persisted into adulthood. Classification as a mature teratoma carries with it a follow-up regimen that includes numerous radiological investigations with their attendant radiation exposure. A positive histological diagnosis and separate classification of adult dermoid tumours would allay clinical fears of recurrence and metastasis and negate the need for repeated radiological investigations.
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Teratoma/patología , Neoplasias Testiculares/patología , Adulto , Humanos , MasculinoRESUMEN
1 The effect of thiopentone, methohexitone, urethane and ketamine on the uptake and release of gamma-aminobutyric acid (GABA) and D-aspartate by rat thalamic slices has been investigated. 2 A high, supra-anaesthetic concentration of methohexitone increased the uptake of both D-aspartate and GABA. 3 None of the anaesthetics used had any detectable effect upon the spontaneous release of either amino acid. 4 Urethane and ketamine had no effect upon the K+-stimulated release of either amino acid. 5 Methohexitone and thiopentone produced a biphasic dose-response on the K+-stimulated release of both amino acids; low concentrations enhanced release, high concentrations depressed release. 6 Bicuculline hydrochloride and picrotoxin both significantly reduced the barbiturate-induced enhancement of K+-stimulated amino acid release, but did not significantly alter the depression of K+-stimulated release at higher barbiturate concentrations. 7 Baclofen, either alone (1 microM to 1 mM), or tested against the barbiturates, had no detectable effect.
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Anestésicos/farmacología , Ácido Aspártico/metabolismo , Neurotransmisores/metabolismo , Tálamo/efectos de los fármacos , Ácido gamma-Aminobutírico/metabolismo , Animales , Baclofeno/farmacología , Bicuculina/farmacología , Calcio/fisiología , Técnicas In Vitro , Masculino , Metohexital/farmacología , Picrotoxina/farmacología , Potasio/farmacología , Ratas , Ratas Endogámicas , Tálamo/metabolismoRESUMEN
Previous angiographic observations have characterized transplant atherosclerosis as a generally diffuse and more distally severe disease with obliteration of secondary branches. However, it has not been firmly established that the disease is structurally and biologically more severe distally. We evaluated this hypothesis with computer-based digitization of subserial segments of the entire perfusion-fixed left anterior descending coronary artery (100 mm Hg for 1 hour with 10% formaldehyde solution) in 25 allografts at autopsy or explant (19 male and 6 female patients; mean age = 50 years, range 16 to 66; mean implant duration = 490 days, range 3 to 1610). The area, thickness, circumference of the intima and media, and the relative and absolute luminal narrowing were evaluated in a mean of 10 left anterior descending coronary artery sections per allograft. The percentage of luminal narrowing (intimal area/[intimal area + luminal area] x 100) was similar between proximal and distal segments of the left anterior descending coronary artery (45% versus 41%, p > 0.05), and the mean absolute intimal thicknesses (in millimeters) of proximal and distal segments of the left anterior descending coronary artery also were not different (0.32 versus 0.22, p > 0.05). In addition, the 95% confidence intervals for intimal thicknesses of proximal and distal segments were comparable. Because the absolute arterial size of proximal segments is naturally larger than that of distal segments (external diameter 9.37 versus 6.79, p < 0.0001), an appearance of progressive tapering may be visualized angiographically, even though the biologic severity of the disease is geographically uniform. Similarly, observations of obliterated secondary branches in distal segments may result from naturally smaller distal luminal areas which may be occluded by less intimal thickening than would be required proximally. These data emphasize that transplant atherosclerosis is biologically uniform from proximal to distal locations. Etiologic and pathogenetic studies on proximal or distal segments should be equally informative.
Asunto(s)
Enfermedad de la Arteria Coronaria/patología , Vasos Coronarios/patología , Trasplante de Corazón/patología , Túnica Íntima/patología , Adolescente , Adulto , Anciano , Estudios de Casos y Controles , Angiografía Coronaria , Enfermedad de la Arteria Coronaria/diagnóstico por imagen , Tejido Elástico/diagnóstico por imagen , Tejido Elástico/patología , Femenino , Humanos , Procesamiento de Imagen Asistido por Computador , Masculino , Persona de Mediana Edad , Procesamiento de Señales Asistido por Computador , Programas Informáticos , Trasplante Homólogo , Túnica Íntima/diagnóstico por imagen , Túnica Media/diagnóstico por imagen , Túnica Media/patologíaRESUMEN
In conclusion, a great deal of indirect and inferential data point to herpesviruses as having a role in atherogenesis. It has been shown that the herpesviruses are able to remain within vascular tissue in a latent state, allowing for reactivation to occur with subsequent sequelae of an active infection. Herpesviruses affect the cellular metabolic activity of cells, induce the accumulation of lipids, and inhibit the production of matrix proteins. They have the ability to inhibit endothelial cell binding to the basement membrane. It is also known that the herpesviruses, particularly CMV, can initiate a variety of immunologic responses that may contribute to endothelial damage, precipitating atherogenesis. We are only beginning to understand how CMV may participate in ACAD. Greater attention must be focused on the exact cause-and-effect relationship between CMV infection and ACAD. Even the presence of CMV genomes in arterial walls of allografts must be viewed conservatively in the knowledge of CMV ubiquity and other probable contributions to ACAD. If CMV is involved in the development of ACAD, as an active or latent infection, directly or indirectly, it probably involves numerous coexistent mechanisms (Figure 5).
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Enfermedad de la Arteria Coronaria/etiología , Infecciones por Citomegalovirus/complicaciones , Endotelio Vascular , Trasplante de Corazón , Herpesviridae/patogenicidad , Complicaciones Posoperatorias , Infecciones por Citomegalovirus/inmunología , Herpesviridae/inmunología , Herpesviridae/metabolismo , Humanos , Metabolismo de los Lípidos , Microscopía Electrónica , Trasplante HomólogoRESUMEN
AIMS: To describe the histological features of the liver in patients with a Fontan circulation. METHODS: Specimens from liver biopsies carried out as part of preoperative assessment prior to extracardiac cavopulmonary conversion of an older style Fontan were examined and scored semi-quantitatively for pertinent histological features. To support the use of the scoring, biopsy specimens were also ranked by eye for severity to allow correlation with assigned scores. RESULTS: Liver biopsy specimens from 18 patients with a Fontan circulation were assessed. All specimens showed sinusoidal fibrosis. In 17 cases there was at least fibrous spur formation, with 14 showing bridging fibrosis and 2 showing frank cirrhosis. In 17 cases at least some of the dense or sinusoidal fibrosis was orcein positive, although a larger proportion of the dense fibrous bands were orcein positive compared with the sinusoidal component. All specimens showed marked sinusoidal dilatation, and 14 showed bile ductular proliferation; 1 showed minimal iron deposition, and 1 showed mild lobular lymphocytic inflammation. There was no cholestasis or evidence of hepatocellular damage. Similar appearances were observed in 2 patients with severe tricuspid regurgitation. DISCUSSION: The histological features of the liver in patients with a Fontan circulation are similar to those described in cardiac sclerosis. Sinusoidal dilatation and sinusoidal fibrosis are marked in the Fontan series. The presence of a significant amount of orcein negative sinusoidal fibrosis suggests there may be a remediable component, although the dense fibrous bands are predominantly orcein positive, suggesting chronicity and permanence. No inflammation or hepatocellular damage is evident, suggesting that fibrosis may be mediated by a non-inflammatory mechanism.
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Procedimiento de Fontan/efectos adversos , Cirrosis Hepática/patología , Biopsia , Dilatación Patológica/etiología , Dilatación Patológica/patología , Humanos , Hígado/patología , Cirrosis Hepática/etiología , Cirrosis Hepática/metabolismo , Oxazinas/metabolismo , Reoperación , Insuficiencia de la Válvula Tricúspide/patologíaRESUMEN
The release of neutrophil chemotactic activity by the guinea pig alveolar macrophage (AM) is dependent on the fifth component of complement (C5) on the cell surface. Because one potent chemotactic factor released by AMs is leukotriene B4 (LTB4), we hypothesized that cell surface C5 may modulate LTB4 release. To test this hypothesis, human AMs obtained by bronchoalveolar lavage from 12 subjects were cultured for 4 hours in the presence of anti-C5 Fab' antibodies with stimuli. The cultures were harvested and evaluated for LTB4 by radioimmunoassay. The LTB4 levels in supernatants obtained from AMs cultured in media alone were variable (447 +/- 63 pg/ml), but the levels were increased when AMs were cultured with the stimuli-opsonized zymosan, immune complexes, or lipopolysaccharide (233%, 49%, and 114% increase, respectively, compared with macrophages cultured in media alone, p less than 0.05). Culturing the AMs with anti-C5 Fab' antibodies inhibited the release of LTB4 induced by opsonized zymosan, immune complexes, or lipopolysaccharide (78%, 41%, and 82% inhibition, respectively, p less than 0.05). Consistent with these observations, anti-C5 Fab' antibodies also decreased the neutrophil chemotactic activity of culture supernatants obtained from AMs stimulated with the same stimuli (p less than 0.001). These data suggest that AM release of LTB4 may be C5-dependent.
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Complemento C5/fisiología , Leucotrieno B4/metabolismo , Macrófagos/metabolismo , Anticuerpos/farmacología , Complejo Antígeno-Anticuerpo/farmacología , Líquido del Lavado Bronquioalveolar/citología , Células Cultivadas , Complemento C5/inmunología , Humanos , Fragmentos Fab de Inmunoglobulinas/farmacología , Lipopolisacáridos/farmacología , Activación de Macrófagos/efectos de los fármacos , Proteínas Opsoninas , Alveolos Pulmonares/citología , Zimosan/farmacologíaRESUMEN
Defective regulation of neutrophil chemotaxis occurs in patients with alcoholic liver disease. One potent mediator of neutrophil chemotaxis is the complement-derived neutrophil chemoattractant, C5a, which can be inhibited by a serum protein, chemotactic factor inactivator. We hypothesized that chemotactic factor inactivator elevation might, in part, explain the defective neutrophil chemotaxis seen in patients with alcoholic hepatitis. To test this hypothesis, sera were collected from 22 patients with alcoholic hepatitis and 9 normal controls, and evaluated for the antigenic presence of chemotactic factor inactivator using an ELISA test. Chemotactic factor inactivator levels were found to be markedly elevated in patients with alcoholic hepatitis (162 +/- 24 micrograms per ml) compared to normals (60 +/- 3 micrograms per ml, p less than 0.01). Subdividing the hepatitis patients revealed that the elevation of chemotactic factor inactivator was found to be greatest in those patients with mild alcoholic hepatitis (prothrombin time within normal limits and bilirubin less than or equal to 5 mg per dl, 256 +/- 44 micrograms per ml, p less than 0.001), while the group with the severest hepatic dysfunction (prolonged prothrombin time and bilirubin greater than 5 mg per dl) did not differ significantly from controls (71 +/- 11 micrograms/ml, p less than 0.2). Importantly, the inhibition of C5a-induced chemotactic activity by partially purified chemotactic factor inactivator correlated with antigenic amounts of chemotactic factor inactivator in serum (r = 0.63, p less than 0.05). The C5a inhibitory activity in sera obtained from patients with alcoholic hepatitis coprecipitated with chemotactic factor inactivator when serum was precipitated by ammonium sulfate precipitation (45 to 64% saturation).(ABSTRACT TRUNCATED AT 250 WORDS)
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Aminopeptidasas/sangre , Factores Quimiotácticos/antagonistas & inhibidores , Hepatopatías Alcohólicas/sangre , Antígenos/análisis , Bilirrubina/sangre , Factores Quimiotácticos/sangre , Factores Quimiotácticos/inmunología , Quimiotaxis de Leucocito , Cromatografía de Afinidad , Complemento C5/análisis , Complemento C5a , Ensayo de Inmunoadsorción Enzimática , Hepatitis Alcohólica/sangre , Hepatitis Alcohólica/clasificación , Hepatitis Alcohólica/inmunología , Humanos , Inmunoglobulina A/análisis , Inmunoglobulina G/análisis , Neutrófilos , Tiempo de ProtrombinaRESUMEN
Several proteins have been described that can modulate the activity of the complement component C5a, a potent chemoattractant for neutrophils. One of these inhibitors has been termed chemotactic factor inactivator (CFI). We hypothesized that CFI was antigenically present in normal human serum and that antigenic levels would correlate with the ability of serum to inhibit C5a. To test this hypothesis, CFI was purified from normal human serum, antibodies to CFI were developed in rabbits, and these reagents were used to develop an enzyme-linked immunoadsorbent assay to measure CFI. Sera from 32 normal volunteers were assayed for CFI and found to contain 77 +/- 29 micrograms/ml (range 17 to 137 micrograms/ml). Partially purified CFI from normal human sera was found to inhibit 61% +/- 9% (range 45% to 75%) of the ability of C5a to attract human neutrophils. Importantly, the ability of CFI to inhibit C5a-induced neutrophil chemotaxis correlated with the antigenic amounts of CFI (r = 0.68, P less than 0.05), suggesting that CFI is a major inhibitor of C5a. This was confirmed by the finding that (1) all C5a inhibitory activity coprecipitated with CFI by ammonium sulfate precipitation (45% to 65% saturation), and (2) depletion of this ammonium sulfate fraction of CFI resulted in a major loss in its ability to inhibit the chemotactic activity of C5a (57% vs. 8% inhibition, P less than 0.01). To determine whether CFI could play a role in the modulation of inflammation at tissue sites, normal bronchoalveolar lavage fluid was evaluated for the presence of CFI. CFI was identified in all fluids (mean 0.50 +/- 0.09 micrograms/mg albumin, range 0.14 to 1.43 micrograms/mg albumin.(ABSTRACT TRUNCATED AT 250 WORDS)
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Aminopeptidasas , Factores Quimiotácticos/antagonistas & inhibidores , Complemento C5/antagonistas & inhibidores , Adulto , Factores Quimiotácticos/análisis , Factores Quimiotácticos/farmacología , Complemento C5a , Electroforesis en Gel de Poliacrilamida , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Inflamación/inmunología , Pulmón/análisis , Masculino , Irrigación TerapéuticaRESUMEN
The contribution of specific risk factors to the development of coronary arteriopathy in human heart allografts remains unclear. Allografts from 15 patients, 11 males and 4 females, aged 15 to 58 years (mean, 40 years), with patient survival from 0.5 to 24 months (mean, 8.6 months) with "triple drug therapy," had the entire coronary artery trees removed, with 184 4-mm arterial segments studied. Luminal narrowing was measured by means of digitization on a video image analysis system, and extent of luminal narrowing (cross-sectional area reduction: [Intimal area/Intimal area + Luminal area] X 100 = %) was related to 40 individual risk factors, including demographic, hemodynamic, immune, environmental, and therapeutic factors. Mean luminal narrowing, considering all coronary segments, was significantly greater in patients with higher versus lower mean cholesterol levels (246 vs 163 mg/dl), triglyceride levels (328 vs 145 mg/dl), and body mass indices (31 vs 22 kg/m2) at 62% versus 38%, 59% versus 42% and 61% versus 44% luminal narrowing, respectively. Considering all coronary segments from all heart allografts, mean luminal narrowing steadily progressed with duration of implant, reaching greater than 60% within 6 months. Mean luminal narrowing was identical in proximal and distal halves of coronary trees at 51% and 50%, respectively. Rejection episodes, considering all degrees of rejection, were strongly related to percent luminal narrowing (p = 0.01). Multivariate analysis indicated the single most predictive risk factor to be posttransplant body mass index (r = 0.77; p = 0.0009).(ABSTRACT TRUNCATED AT 250 WORDS)
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Enfermedad Coronaria/epidemiología , Trasplante de Corazón , Hiperlipidemias/epidemiología , Obesidad/epidemiología , Adulto , Índice de Masa Corporal , Enfermedad Coronaria/patología , Vasos Coronarios/patología , Femenino , Rechazo de Injerto , Trasplante de Corazón/mortalidad , Trasplante de Corazón/patología , Humanos , Inmunosupresores/uso terapéutico , Masculino , Análisis Multivariante , Factores de RiesgoRESUMEN
The degree to which transplant arteriopathy in solid organ allografts is an atheromatous process remains somewhat controversial. If atheromata develop as common and integral components of the arteriopathic lesions, then the process may be approached therapeutically in a manner analogous to native atheromatous diseases. Approaches to understanding the arteriopathic process may include not only the modulation of alloimmunity, but also the interruption of "storage" phenomena. We have examined the epicardial coronary arteries of nearly 50 explanted human heart allografts using biochemical, morphological, morphometrical, immunohistochemical, and molecular techniques in an effort to establish the degree, nature, and distribution of lipid accumulation in the vessel walls. Concomitantly, we studied the ascertainment of proteoglycan gene expression, represented by biglycan and decorin messenger RNA, and the localization of proteoglycan proteins in the vessels. The degree of lipid and proteoglycan buildup in both the intima and media of transplanted vessels is striking, and correlated strongly with intimal thickening, cross-sectional area reduction of the lumen, cumulative cyclosporine dose, corticosteroid dose, and serum cholesterol levels. Notably, lipid accumulation is not related to implant duration, this being true in an unselected series of "failed" allografts ranging from 4 to 1610 days post-transplant. The profound lipid accumulation in coronary walls of many grafts begins very early post-transplant and appears to contribute substantially to intimal thickening. Whether dysregulation of proteoglycan production leads to entrapment of lipids and lipoproteins remains an important and testable hypothesis.
Asunto(s)
Enfermedad de la Arteria Coronaria/metabolismo , Rechazo de Injerto/metabolismo , Trasplante de Corazón/fisiología , Metabolismo de los Lípidos , Proteoglicanos/metabolismo , Enfermedad Crónica , Enfermedad de la Arteria Coronaria/microbiología , Enfermedad de la Arteria Coronaria/patología , Vasos Coronarios/metabolismo , Vasos Coronarios/microbiología , Vasos Coronarios/patología , Citomegalovirus/aislamiento & purificación , Tejido Elástico/metabolismo , Tejido Elástico/patología , Expresión Génica , Rechazo de Injerto/microbiología , Rechazo de Injerto/patología , Trasplante de Corazón/patología , Humanos , Proteoglicanos/genética , Factores de RiesgoRESUMEN
Endocardial infiltrates (EI) are a common and often problematic observation in endomyocardial biopsy specimens (EMBs) from patients receiving cyclosporine immunosuppression following cardiac transplant. Histologic and immunohistologic findings in 23 EMBs from 19 patients and 15 autopsy or explanted allografts demonstrated EIs to be rich in B lymphocytes (871/mm2) compared to T-lymphocytes (803/mm2). Macrophages also demonstrated an endocardial preference over deeper myocardium. In contrast, T-lymphocytes outnumbered B-lymphocytes in deeper myocardium (mean 44/mm2 versus 22/mm2) especially when rejection was present. In allograft specimens, the overall number of typical nodular EIs or percent length of endocardial involvement by EI did not correlate with the presence or absence of myocardial rejection at autopsy or explant but were related to implant duration (r = +0.63, p less than 0.01) and number of previous rejection episodes. The number of thin, nondiscrete endocardial infiltrates was greater in hearts with any myocardial rejection or inflammation present. No relationship was observed between EIs present in either EMB or allografts and the cumulative or mean dose or mean serum level of cyclosporine. Thus, a distinct morphologic and immunohistologic profile distinguishes EIs from acute rejection.
Asunto(s)
Linfocitos B/patología , Endocardio/patología , Rechazo de Injerto , Trasplante de Corazón/patología , Adolescente , Adulto , Anciano , Antígenos CD/análisis , Linfocitos B/inmunología , Movimiento Celular , Niño , Ciclosporina/uso terapéutico , Endocardio/inmunología , Femenino , Rechazo de Injerto/efectos de los fármacos , Trasplante de Corazón/inmunología , Antígenos de Histocompatibilidad/análisis , Humanos , Inmunohistoquímica , Inmunofenotipificación , Antígenos Comunes de Leucocito , Leucocitos/inmunología , Leucocitos/patología , Macrófagos/inmunología , Macrófagos/patología , Masculino , Persona de Mediana Edad , Miocardio/inmunología , Miocardio/patología , Linfocitos T/inmunología , Linfocitos T/patología , Trasplante HomólogoRESUMEN
Transplant arteriopathy is a major late complication in human heart allograft recipients and the pathogenesis of such arteriopathy remains uncertain. The degree to which lipids and atheromata are involved in the arteriopathic lesions remains unsettled, and there is uncertainty regarding the significance of insudation or retention of lipids within the coronary artery walls of transplanted hearts. On current immunosuppressive regimens, most patients experience an increased serum total cholesterol and low-density lipoprotein cholesterol after transplant. Elevation of these blood lipids has an undetermined relationship to arteriopathy. We carried out morphological, morphometric, immunohistochemical, ultrastructural, and biochemical studies of particular coronary artery segments from 23 unselected explant or autopsy allografts and donor age-matched native coronary controls. Patients died of cardiac and non-cardiac reasons over a period of 4 to 1610 days after transplant. Atheromata were frequent, and diffuse intra- and extra-cellular accumulation of lipids in both intimal and medial walls was documented by oil red O positivity, immunohistochemical staining (muscle-specific alpha-actin), transmission and scanning electron microscopy, and biochemical analysis. Mean total cholesterol, esterified cholesterol, free cholesterol, and phospholipid content (microgram/cm2 intimal surface area) and concentration (microgram/mg dry defatted weight) in arteriopathic coronaries were > 10-fold higher than in comparable native coronary segments. Extent of lipids in the arterial walls was highly correlated with digitized percent luminal narrowing, mean daily and cumulative cyclosporin dose, and mean cumulative prednisone dose. Our data suggests strongly that lipid accumulation is an important early and persistent phenomenon in the development of transplant arteriopathy.
Asunto(s)
Vasos Coronarios/metabolismo , Trasplante de Corazón , Metabolismo de los Lípidos , Enfermedades Vasculares/etiología , Adolescente , Adulto , Anciano , Arterias , Vasos Coronarios/patología , Femenino , Humanos , Masculino , Microscopía Electrónica , Persona de Mediana Edad , Complicaciones Posoperatorias , Trasplante HomólogoRESUMEN
In heart transplantation, long-term engraftment success is severely limited by the rapid development of obliterative disease of the coronary arteries. Data from various groups have been suggestive of a pathogenetic role of herpesviruses, particularly human cytomegalovirus, in accelerated allograft coronary artery disease; however, results are not yet conclusive. This study examines the hypothesis that human cytomegalovirus infection of allograft tissues is related pathogenetically and directly to accelerated coronary artery disease. Using in situ DNA hybridization and polymerase chain reaction, we examined particular coronary artery segments from 41 human heart allografts (ranging from 4 days to greater than 4 years after transplantation; mean, 457 days) and 22 donor age- and gender-comparable, coronary site-matched trauma victims for presence of human cytomegalovirus DNA. Human cytomegalovirus genome was detected in 8 of 41 (19.5%) allografts and in 1 of 22 (4.5%) control hearts. This difference in positivity was not statistically significant (P = 0.10). In the human cytomegalovirus-positive hearts, viral genome was localized to perivascular myocardium or coronary artery media or adventitia. Human cytomegalovirus genome was not detected in arterial intima of any allograft or control heart, although human cytomegalovirus genome was readily identified within intima of small pulmonary arteries from lung tissue with human cytomegalovirus pneumonitis. By statistical analyses, the presence of human cytomegalovirus genome was not associated with the nature or digitized extent of transplant arteriopathy, evidence of rejection, allograft recipient or donor serological data suggestive of human cytomegalovirus infection, duration of allograft implantation, or causes of death or retransplantation. Thus, our data indicate a low frequency of detectable human cytomegalovirus genome in accelerated coronary artery disease and do not support a direct role for human cytomegalovirus in vascular wall infection or in the development of accelerated coronary artery disease.