RESUMEN
Integrating clinical pathology data with anatomic pathology data is a common practice when reporting findings in the context of nonclinical toxicity studies and aids in understanding and communicating the nonclinical safety profile of test articles in development. Appropriate pathology data integration requires knowledge of analyte and tissue biology, species differences, methods of specimen acquisition and analysis, study procedures, and an understanding of the potential causes and effects of a variety of pathophysiologic processes. Neglecting these factors can lead to inappropriate data integration or a missed opportunity to enhance understanding and communication of observed changes. In such cases, nonclinical safety information relevant to human safety risk assessment may be misrepresented or misunderstood. This "Points to Consider" manuscript presents general concepts regarding pathology data integration in nonclinical studies, considerations for avoiding potential oversights and errors in data integration, and focused discussion on topics relevant to data integration for several key organ systems including liver, kidney, and cardiovascular system.
Asunto(s)
Patología Clínica , Toxicología , Humanos , Patología Clínica/métodos , Políticas , Medición de Riesgo , Toxicología/métodosRESUMEN
Detection of test article-related effects and the determination of the adversity of those changes are the primary goals of nonclinical safety assessment studies for drugs and chemicals in development. During these studies, variables that are not of primary interest to investigators may change and influence data interpretation. These variables, often referred to as "nuisance factors," may influence other groups of data and result in "block or batch effects" that complicate data interpretation. Definitions of the terms "nuisance factors," "block effects," and "batch effects," as they apply to nonclinical safety assessment studies, are reviewed. Multiple case examples of block and batch effects in safety assessment studies are provided, and the challenges these bring to pathology data interpretation are discussed. Methods to mitigate the occurrence of block and batch effects in safety assessment studies, including statistical blocking and utilization of study designs that minimize potential confounding variables, incorporation of adequate randomization, and use of an appropriate number of animals or repeated measurement of specific parameters for increased precision, are reviewed. [Box: see text].
Asunto(s)
Evaluación Preclínica de Medicamentos , Animales , Humanos , Políticas , Proyectos de InvestigaciónRESUMEN
Emergency providers at Orlando Regional Medical Center in Orlando. FL, faced multiple challenges in responding to the worst mass shooting in U.S. history. As the scene of the shooting was only three blocks away from the hospital, there was little time to prepare when notified that victims would begin arriving shortly after 2 a.m. on June 12. Also, fears of a gunman near the hospital briefly put the ED on lock down. However, using the incident command system, the hospital was able to mobilize quickly, receiving 44 patients, nine of whom died shortly after arrival. Administrators note that recent training exercises geared toward a mass shooting event facilitated the response and probably saved lives. Patients arrived at the hospital in two waves, with the initial surge occurring right after the hooting took place around 2 a.m., and the second surge occurring about three hours later. At one point, more than 90 patients were in the ED, more than half for reasons unrelated to the shooting. Clinicians contended with a much higher than usual noise level while treating patients, making it hard to hear reports from EMS personnel. Also, treatment had to commence prior to identification for some patients who arrived unconscious or unable to speak. While surgeons and other key specialists were called into the hospital to address identified needs, administrators actually called hospital personnel to tell them not to come in unless they were notified. This prevented added management hurdles.
Asunto(s)
Planificación en Desastres , Incidentes con Víctimas en Masa , Entrenamiento Simulado , Centros Traumatológicos/organización & administración , Heridas por Arma de Fuego/terapia , Aglomeración , Florida , HumanosRESUMEN
The mechanisms initiating eosinophil influx into sites of inflammation have been well studied in allergic disease but are poorly understood in other settings. This study examined the roles of TLR2 and mast cells in eosinophil accumulation during a nonallergic model of eosinophilia-associated colitis. TLR2-deficient mice (TLR2(-/-)) developed a more severe colitis than wild-type mice in the dextran sodium sulfate (DSS) model. However, they had significantly fewer eosinophils in the submucosa of the cecum (P < 0.01) and mid-colon (P < 0.01) than did wild-type mice after DSS treatment. Decreased eosinophilia in TLR2(-/-) mice was associated with lower levels of cecal CCL11 (P < 0.01). Peritoneal eosinophils did not express TLR2 protein, but TLR2 ligand injection into the peritoneal cavity induced local eosinophil recruitment, indicating that TLR2 activation of other cell types can mediate eosinophil recruitment. After DSS treatment, mast cell-deficient (Kit(W-sh/W-sh)) mice had similar levels of intestinal tissue eosinophilia were observed as those in wild-type mice. However, mast cell-deficient mice were partially protected from DSS-induced weight loss, an effect that was reversed by mast cell reconstitution. Overall, this study indicates a critical role for indirect TLR2-dependent pathways, but not mast cells, in the generation of eosinophilia in the large intestine during experimental colitis and has important implications for the regulation of eosinophils at mucosal inflammatory sites.
Asunto(s)
Colitis Ulcerosa/inmunología , Enfermedad de Crohn/inmunología , Eosinofilia/genética , Eosinofilia/inmunología , Receptor Toll-Like 2/inmunología , Animales , Colitis Ulcerosa/inducido químicamente , Enfermedad de Crohn/inducido químicamente , Sulfato de Dextran/toxicidad , Modelos Animales de Enfermedad , Eosinofilia/patología , Eosinófilos/inmunología , Eosinófilos/patología , Recuento de Leucocitos , Mastocitos/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Mutantes , Receptor Toll-Like 2/genéticaRESUMEN
BACKGROUND: The etiology of inflammatory bowel diseases (IBD) is largely unknown, but appears to be perpetuated by uncontrolled responses to antigenic components of the endogenous flora. Tolerance to antigenic stimulation can be achieved by exposure to a given antigen in high amounts (high dose tolerance). Colitis induced by feeding of Dextran Sodium Sulfate (DSS) is an often-used animal model mimicking clinical and histological features of human IBD. AIMS: We investigated whether treatment with high doses of endogenous bacterial components can affect the response to these antigenic components and thus impact the course of the inflammatory response induced by DSS. METHODS: 129/SvEv mice were injected intravenously in the tail vein with lysates prepared from fecal material of conventionally-raised mice. Control mice received a solution of bacterial antigen-free lysates prepared from fecal material of germ-free mice. Seven days later, colitis was induced in these mice by introducing DSS (3.5%) in the drinking water for 5 days. Onset and course of the inflammatory response was monitored by assessment of weight loss. Mice were sacrificed at day 7 post colitis induction and tested for histopathologic injury, intestinal cytokine release, and systemic response to bacterial antigens. RESULTS: Intravenous injection with fecal lysates reduced intestinal and antigen-stimulated systemic pro-inflammatory cytokine release and prevented DSS-induced weight loss and intestinal injury. CONCLUSION: Pretreatment with high amount of endogenous bacterial components has a profound tolerogenic effect on the systemic and mucosal immune responses resulting in reduced intestinal inflammation and abrogates colitis-induced weight loss.
Asunto(s)
Colitis/inmunología , Colitis/terapia , Colon/microbiología , Animales , Colitis/inducido químicamente , Colon/inmunología , Citocinas/metabolismo , Sulfato de Dextran/efectos adversos , Heces/química , Inyecciones Intravenosas , Recuento de Linfocitos , Ratones , Ratones Endogámicos , Linfocitos T/inmunología , Linfocitos T Reguladores , Pérdida de Peso/inmunologíaRESUMEN
Integrating clinical pathology data with anatomic pathology data is a common practice when reporting findings in the context of nonclinical toxicity studies and aids in understanding and communicating the nonclinical safety profile of test articles in development. Appropriate pathology data integration requires knowledge of analyte and tissue biology, species differences, methods of specimen acquisition and analysis, study procedures, and an understanding of the potential causes and effects of a variety of pathophysiologic processes. Neglecting these factors can lead to inappropriate data integration or a missed opportunity to enhance understanding and communication of observed changes. In such cases, nonclinical safety information relevant to human safety risk assessment may be misrepresented or misunderstood. This "Points to Consider" manuscript presents general concepts regarding pathology data integration in nonclinical studies, considerations for avoiding potential oversights and errors in data integration, and focused discussion on topics relevant to data integration for several key organ systems, including liver, kidney, and cardiovascular systems.
Asunto(s)
Patología Clínica , Toxicología , Animales , Evaluación Preclínica de Medicamentos/métodos , Evaluación Preclínica de Medicamentos/veterinaria , Humanos , Patología Clínica/métodos , PolíticasRESUMEN
Impaired epithelial barrier function and estrogens are recognized as factors influencing inflammatory bowel disease (IBD) pathology and disease course. Estrogen receptor-ß (ERß) is the most abundant estrogen receptor in the colon and a complete absence of ERß expression is associated with disrupted tight-junction formation and abnormal colonic architecture. The aim of this study was to determine whether ERß signaling has a role in the maintenance of epithelial permeability in the colon. ERß mRNA levels and colonic permeability were assessed in IL-10-deficient mice and HLA-B27 rats by RT-PCR and Ussing chambers. ERß expression and monolayer resistance were measured in HT-29 and T84 colonic epithelial monolayers by RT-PCR and electric cell-substrate impedance sensing. The effect of 17ß-estradiol and an estrogen agonist [diarylpropionitrile (DPN)] and antagonist (ICI 182780) on epithelial resistance in T84 cells was measured. Expression of ERß and proinflammatory cytokines was investigated in colonic biopsies from IBD patients. Levels of ERß mRNA were decreased, whereas colonic permeability was increased, in IL-10-deficient mice and HLA-B27 transgenic rats prior to the onset of colitis. T84 cells demonstrated higher resistance and increased levels of ERß mRNA compared with HT-29 cells. 17ß-estradiol and DPN induced increased epithelial resistance in T84 cells, whereas an ERß blocker prevented the increased resistance. Decreased ERß mRNA levels were observed in colonic biopsies from IBD patients. This study suggests a potential role for ERß signaling in the modulation of epithelial permeability and demonstrates reduced ERß mRNA in animal models of colitis and colon of patients with inflammatory bowel disease.
Asunto(s)
Colon/metabolismo , Células Epiteliales/metabolismo , Receptor beta de Estrógeno/metabolismo , Inflamación/metabolismo , Mucosa Intestinal/metabolismo , Análisis de Varianza , Animales , Línea Celular , Células Cultivadas , Colon/efectos de los fármacos , Células Epiteliales/citología , Células Epiteliales/efectos de los fármacos , Estradiol/análogos & derivados , Estradiol/farmacología , Antagonistas de Estrógenos/farmacología , Receptor beta de Estrógeno/agonistas , Receptor beta de Estrógeno/genética , Femenino , Fulvestrant , Antígeno HLA-B27/genética , Antígeno HLA-B27/metabolismo , Humanos , Inflamación/genética , Interleucina-10/genética , Interleucina-10/metabolismo , Mucosa Intestinal/efectos de los fármacos , Masculino , Ratones , Ratones Noqueados , Nitrilos/farmacología , Permeabilidad , Propionatos/farmacología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Ratas , Ratas TransgénicasRESUMEN
Age-associated changes in immune function and their implications for intestinal inflammation are poorly understood. Defects in innate immunity have been shown to enhance intestinal inflammation and have been demonstrated upon aging. This study aimed to determine the consequences of aging in the presence and absence of TLR2 on intestinal inflammation. Young and aged (>60 weeks), control C57Bl/6 and TLR2-deficient (TLR2(-/-)) mice were examined. The cecum and mid-colon were analyzed for tissue damage, cytokine profiles, and trefoil factor 3 (TFF3) expression at baseline or after 5 days of treatment with dextran sodium sulfate (DSS) and 5 or 13 days recovery. Untreated, aged TLR2(-/-) mice had no significant intestinal inflammation but had reduced colonic IFN-gamma and IL-10 compared with younger mice. Aged TLR2(-/-) mice developed more severe colitis than other groups, as indicated by histological examination and overall weight loss. There were significant increases in colonic IFN-gamma following DSS treatment in young but not in aged mice. TFF3 was substantially reduced in the cecum and increased in the colon of aged but not younger TLR2(-/-) mice following DSS treatment. These results demonstrate that even upon aging, TLR2-deficient animals did not develop intestinal disease. However, they failed to respond appropriately to an inflammatory insult, and the consequences of this were most severe in aged animals. Cytokine and TFF3 changes associated with aging may contribute to more severe intestinal inflammation.
Asunto(s)
Envejecimiento/fisiología , Colitis/fisiopatología , Interferón gamma/fisiología , Receptor Toll-Like 2/deficiencia , Animales , Ciego/efectos de los fármacos , Ciego/patología , Ciego/fisiopatología , Colitis/inducido químicamente , Colitis/genética , Colon/efectos de los fármacos , Colon/patología , Colon/fisiología , Colon/fisiopatología , Sulfato de Dextran/toxicidad , Predisposición Genética a la Enfermedad , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Receptor Toll-Like 2/genética , Receptor Toll-Like 2/fisiologíaRESUMEN
This article aims to validate the factorial structure of the perceived stress scale (PSS10) within the French working population. The analyses conducted confirmed the presence of two distinct factors, interpreted in terms of perceived work overload and perceived personal efficacy. Both factors presented good internal consistency and adequate validity of construct. The authors show and illustrate the predicted link between the two factors and the levels of anxiety and depression. Thus, the PSS 10 is a bi-dimensional scale with satisfactory psychometric proprieties. The results are discussed in the light of their theoretical and practical implications.
Asunto(s)
Estrés Psicológico/diagnóstico , Lugar de Trabajo/psicología , Adulto , Distribución de Chi-Cuadrado , Análisis Factorial , Femenino , Francia , Humanos , Masculino , Persona de Mediana Edad , Psicometría , MuestreoRESUMEN
BACKGROUND: Thrombin generation assays (TGA) have potential applications as measures of hemostatic balance in animal models. However, variations in plasma processing greatly influence human TGA, and may also impact on the translational value of TGA in animal studies. OBJECTIVES: The purpose of the study was to compare the performance characteristics of Sprague-Dawley rat plasma prepared by single vs double centrifugation protocols in TGA and fibrinolysis assays. METHODS: Platelet-poor plasma (PPP) from adult rats (n = 20 males; 20 females) was prepared by centrifugation at 1200g × 12 min, or 2 sequential centrifugations of 2500g × 15 min. Plasma aliquots were assayed fresh and after freeze-thaw in a commercial fluorogenic TGA (Technothrombin TGA, Technoclone) using 2 different trigger reagents containing approximately 7 pM human tissue factor. In addition to TGA variables (lag time, peak thrombin, endogenous thrombin potential), we compared clotting time test and fibrinogen concentration, residual platelet and platelet-derived microparticle (PMP) counts measured by flow cytometry, and variables of fibrin clot formation and lysis measured in turbidimetric assays. RESULTS: Single-centrifugation PPP demonstrated significantly greater thrombin-generating potential regardless of trigger reagent, yielded higher residual platelet and procoagulant PMP counts, and more stable fibrin clot profiles. The influence of a freeze-thaw cycle on TGA varied depending on trigger reagent, and male sex was associated with an overall "procoagulant" phenotype. CONCLUSIONS: Preanalytic processing and sex have significant effects on many functional measures of hemostasis in rats. A standardized double centrifugation protocol to prepare PPP is recommended for future studies.
Asunto(s)
Recolección de Muestras de Sangre/veterinaria , Fibrinólisis , Ratas Sprague-Dawley/sangre , Trombina/metabolismo , Animales , Plaquetas/metabolismo , Femenino , Citometría de Flujo/veterinaria , Congelación , Masculino , Nefelometría y Turbidimetría/veterinaria , Ratas/sangre , Trombina/análisisRESUMEN
BACKGROUND: There is a paucity of information regarding cardiac troponin (cTn) concentrations in peripheral blood of nonhuman primates (NHP). Even less is known regarding cTn concentrations in monkeys that are restrained for oral or intravenous (iv) dosing. OBJECTIVES: The objectives of these studies were to (1) determine cardiac troponin I (cTnI) concentration in resting Cynomolgus monkeys and investigate biologic variability in cTnI concentration over time, (2) determine cTnI changes in restrained monkeys given sham oral dosing, and (3) determine cTnI changes in restrained NHP given a sham intravenous dosing. METHODS: The Research Use Only Erenna cTnI ultrasensitive immunoassay based on single molecule counting technology was used to determine serum cTnI concentration in longitudinal studies of male Cynomolgus monkeys at rest, and after sham oral and intravenous dosing. Animals were catheterized prestudy, and blood samples were collected by an automated sampling device to limit disturbance of the animals during studies. RESULTS: In resting monkeys cTnI concentrations were relatively low and constant and ranged from 0.2 to 9.6 pg/mL (mean = 2.5 pg/mL), with minimal variability during a 24-hour period. Animals given sham oral dosing also had low cTnI concentration with little variability similar to the resting values. Several animals restrained for intravenous dosing had a small transient increase in cTnI concentration (~5-25 pg/mL) that resolved quickly within one to 3 hours postinjection. CONCLUSIONS: Results of this longitudinal study provide information that may be important in differentiating effects of animal handling from those associated with compound-related effects in preclinical toxicology studies of drugs in development.
Asunto(s)
Troponina I/sangre , Administración Intravenosa/veterinaria , Animales , Inmunoensayo/veterinaria , Estudios Longitudinales , Macaca fascicularis , Masculino , Miocardio/metabolismo , Restricción Física/veterinaria , Sensibilidad y Especificidad , Albúmina Sérica , Troponina I/administración & dosificaciónRESUMEN
BACKGROUND: A critical role for the gut epithelium lies in its ability to discriminate between pathogens and commensals and respond appropriately. Dysfunctional interactions between microbes and epithelia are believed to have a role in inflammatory bowel disease (IBD). In this study, we analyzed microbiota and gene expression in IBD patients and examined responses of mucosal biopsies to bacterial DNA. METHODS: Biopsies were taken from non-inflamed areas of the colon in healthy controls (HC) and Crohn's disease (CD) and ulcerative colitis (UC) patients in remission. Biopsies were snap-frozen or cultured with DNA from Lactobacillus plantarum (LP) or Salmonella dublin (SD). Gene expression was analyzed under basal conditions and in response to DNA. Gene networks were analyzed using Ingenuity Pathways software. Mucosal-associated microbiota was analyzed using terminal restriction fragment length polymorphism. Frequency of single nucleotide polymorphisms in NOD2 and TLR9 was assessed. RESULTS: Patients with IBD had altered microbiota, enhanced expression of inflammatory genes, and increased correlations between specific gene expression and microbes. Principle component analysis showed CD and UC patients to cluster independently from healthy controls in both gene expression and microbial analysis. DNA from LP stimulated anti-inflammatory pathways in controls and UC patients, but induced an upregulation of IL17A in CD patients. There were no differences in SNP frequencies of TLR9 or NOD2 in the groups. CONCLUSIONS: Patients with Crohn's disease exhibit altered responses to bacterial DNA. These findings suggest that the gut response to bacterial DNA may depend not only on the specific type of bacterial DNA, but also on the host.
Asunto(s)
Colon/metabolismo , Colon/microbiología , Regulación de la Expresión Génica/efectos de los fármacos , Redes Reguladoras de Genes/genética , Enfermedades Inflamatorias del Intestino/microbiología , Metagenoma/genética , Adulto , Anciano , Anciano de 80 o más Años , Biopsia , Análisis por Conglomerados , Estudios de Cohortes , Colon/patología , Cartilla de ADN/genética , ADN Bacteriano/administración & dosificación , ADN Bacteriano/farmacología , Femenino , Humanos , Lactobacillus plantarum/genética , Masculino , Persona de Mediana Edad , Proteína Adaptadora de Señalización NOD2/genética , Polimorfismo de Longitud del Fragmento de Restricción , Polimorfismo de Nucleótido Simple/genética , Análisis de Componente Principal , Salmonella/genética , Estadísticas no Paramétricas , Receptor Toll-Like 9/genéticaRESUMEN
On exposure to sunlight, urocanic acid (UCA) in the skin is converted from trans to the cis form and distributed systemically where it confers systemic immunosuppression. The aim of this study was to determine if administration of cis-UCA would be effective in attenuating colitis and the possible role of IL-10. Colitis was induced in 129/SvEv mice by administering 5% dextran sodium sulfate (DSS) for 7 days in drinking water. During this period mice received daily subcutaneously injections of cis-UCA or vehicle. To examine a role for IL-10, 129/SvEv IL-10(-/-) mice were injected for 24 days with cis-UCA or vehicle. Clinical disease was assessed by measurement of body weight, stool consistency, and presence of blood. At sacrifice, colonic tissue was collected for histology and measurement of myeloperoxidase and cytokines. Splenocytes were analyzed for CD4+CD25+FoxP3+ T-regulatory cells via flow cytometry. Murine bone-marrow derived antigen-presenting cells were treated with lipopolysaccharide (LPS) ± UCA and cytokine secretion measured. Our results demonstrated that cis-UCA at a dose of 50 µg was effective in ameliorating DSS-induced colitis as evidenced by reduced weight loss and attenuated changes in colon weight/length. This protection was associated with reduced colonic expression of CXCL1, an increased expression of IL-17A and a significant preservation of splenic CD4+CD25+FoxP3+ T-regulatory cells. cis-UCA decreased LPS induced CXCL1, but not TNFα secretion, from antigen-presenting cells in vitro. UCA reduced colonic levels of IFNγ in IL-10(-/-) mice but did not attenuate colitis. In conclusion, this study demonstrates that cis-urocanic acid is effective in reducing the severity of colitis in a chemically-induced mouse model, indicating that pathways induced by ultraviolet radiation to the skin can influence distal sites of inflammation. This provides further evidence for a possible role for sunlight exposure in modulating inflammatory disorders.
Asunto(s)
Colitis/prevención & control , Sulfato de Dextran/efectos adversos , Ácido Urocánico/farmacología , Animales , Células Presentadoras de Antígenos/inmunología , Colitis/inducido químicamente , Citometría de Flujo , Interleucina-10/genética , Interleucina-10/fisiología , Ratones , Ratones NoqueadosRESUMEN
Mutations in toll-like receptors that mediate bacterial recognition by the mammalian innate immune system have the potential to substantially alter the composition of an individual's microbiota. Here we tested this hypothesis by comparing the intestinal microbiota of toll-like receptor 2-deficient mice, both young and middle aged, with that of wild-type mice of the same genetic background, housed together under specific pathogen-free conditions. Bacterial DNA was extracted from mouse caecal tissue samples, amplified using universal bacterial 16S ribosomal RNA gene primers, and cloned into a plasmid vector. Insert-containing colonies were picked for high-throughput sequencing, and sequence data were analysed yielding species-level phylogenetic data. Clone libraries were compared by phylogenetic composition analysis using UniFrac. While pairwise differences in phylogenetic population structure between mutant and wild-type mice were not statistically significant, anosim analysis did demonstrate a significant difference between toll-like receptor 2-deficient mice and their wild-type counterparts. The difference observed was probably due to a high level of colonization of the toll-like receptor 2-deficient mice by two distinct Helicobacter phylotypes that were totally absent from wild-type mice. Principal coordinate analysis clustering indicated that age is a weaker determinate than genotype and maternal heritage in the mouse caecal microbiota. The findings suggest that although mutations in toll-like receptors may cause increased susceptibility to specific opportunistic bacteria, they do not dramatically alter the phylogenetic structure of microbiota.
RESUMEN
Fatty acid binding protein 3 (Fabp3) has been used as a serological biomarker of cardiac injury, but its utility as a preclinical biomarker of injury to skeletal muscle is not well described. Fabp3 concentrations were determined for tissues from Sprague-Dawley rats and found to occur at highest concentrations in cardiac muscle and in skeletal muscles containing an abundance of type I fibers, such as the soleus muscle. Soleus is also a primary site of skeletal muscle (SKM) injury caused by lipid-lowering peroxisome proliferator-activated receptor alpha (PPAR-alpha) agonists. In rats administered repeat doses of a PPAR-alpha agonist, the kinetics and amplitude of plasma concentrations of Fabp3 were consistent with plasma compound concentrations and histopathology findings of swollen, hyalinized, and fragmented muscle fibers with macrophage infiltration. Immunohistochemical detection of Fabp3 revealed focal depletion of Fabp3 protein from injured SKM fibers which is consistent with increased serum Fabp3 concentrations in treated rats. We then assessed the predictivity of serological Fabp3 for SKM necrosis in short duration toxicology studies. Rats were treated with various doses of 27 different compounds, and the predictivity of serological biomarkers was assessed relative to histology in individual rats and in treatment groups. Under these study conditions, Fabp3 was the most useful individual biomarker based on concordance, sensitivity, positive and negative predictive values, and false negative rate. In addition, the combination of Fabp3 and aspartate aminotransferase (AST) had greater diagnostic value than the conventional combination of creatine kinase-MM isoenzyme (CK) and AST.
Asunto(s)
Biomarcadores/metabolismo , Proteínas de Unión a Ácidos Grasos/metabolismo , Músculo Esquelético , Miositis/metabolismo , Xenobióticos/toxicidad , Animales , Anticuerpos Bloqueadores/farmacología , Aspartato Aminotransferasas/metabolismo , Proteína 3 de Unión a Ácidos Grasos , Proteínas de Unión a Ácidos Grasos/genética , Proteínas de Unión a Ácidos Grasos/inmunología , Femenino , Masculino , Fibras Musculares Esqueléticas/metabolismo , Músculo Esquelético/efectos de los fármacos , Músculo Esquelético/metabolismo , Músculo Esquelético/patología , Miocardio/metabolismo , Miositis/inducido químicamente , Miositis/patología , Valor Predictivo de las Pruebas , Ratas , Ratas Sprague-Dawley , Reproducibilidad de los ResultadosAsunto(s)
Bacteriemia/veterinaria , Inflamación/veterinaria , Macaca fascicularis , Enfermedades de los Monos/diagnóstico , Animales , Bacteriemia/diagnóstico , Recuento de Células Sanguíneas/veterinaria , Catéteres de Permanencia/microbiología , Catéteres de Permanencia/veterinaria , Remoción de Dispositivos/veterinaria , Femenino , Inflamación/diagnóstico , Enfermedades de los Monos/sangre , Prótesis e Implantes/efectos adversos , Prótesis e Implantes/microbiologíaRESUMEN
Langerhans cells and mast cells are both resident in large numbers in the skin and act as sentinel cells in host defense. The ability of mast cells to induce Langerhans cell migration from the skin to the draining lymph node in vivo was examined. Genetically mast cell-deficient (W/Wv) mice and control mice were sensitized with IgE Ab in the ear pinna. Seven to 14 days later, mice were challenged with Ag i.v. After a further 18-24 h, epidermal sheets and draining auricular lymph nodes were examined using Langerin/CD207 immunostaining. In mast cell-containing mice, a significant decrease in the number of Langerhans cells was observed at epidermal sites of mast cell activation. A significant increase in total cellularity and accumulation of Langerin-positive dendritic cells was observed in the auricular lymph nodes, draining the sites of IgE-mediated mast cell activation. These changes were not observed in W/Wv mice, but were restored by local mast cell reconstitution. Treatment of mast cell-containing mice with the H2 receptor antagonist cimetidine significantly inhibited the observed IgE/Ag-induced changes in Langerhans cell location. In contrast, Langerhans cell migration in response to LPS challenge was not mast cell dependent. These data directly demonstrate the ability of mast cells to induce dendritic cell migration to lymph nodes following IgE-mediated activation in vivo by a histamine-dependent mechanism.