RESUMEN
The polyclonal stimulation of T cells by bacterial superantigens is involved in the pathogenesis of the toxic shock syndrome in certain staphylococcal and streptococcal infections. Here we describe the onset and kinetics of superantigen-induced cytokine production in situ in spleens of normal BALB/c mice monitored at the level of cytokine mRNA expression by in situ hybridization. Messenger RNAs for interleukin 2 (IL-2), interferon gamma, and tumor necrosis factors (TNF) alpha and beta were not expressed at detectable levels in spleens of unstimulated animals but became visible already 30 min after intraperitoneal application of 50 micrograms staphylococcal enterotoxin B. All mRNA levels showed peak expression approximately 3 h after injection and a slow decrease up to 24 h after injection. Expression of the mRNAs was restricted to the T cell-dependent area of the periarteriolar lymphatic sheets of the spleen. Interestingly, TNF-alpha mRNA showed a biphasic response, the early appearing mRNA had the same localization as the other mRNAs, whereas after 3 h TNF-alpha mRNA showed a broader distribution indicating a second cell population producing TNF-alpha. The expression of IL-2 and TNF proteins in the serum increased in parallel to the observed mRNA changes with a slight delay. The presence of macrophages was not required for the expression of the cytokine mRNAs in the spleen as the expression was unchanged in macrophage-depleted mice. Only the second phase of TNF-alpha mRNA expression was abrogated in such animals. The expression of all mRNAs was completely suppressed by prior administration of cyclosporin A. These data show that nonphagocytic cells are the essential superantigen-presenting cells in vivo and indicate that at least part of the pathogenetic TNF-alpha is T cell derived.
Asunto(s)
Citocinas/biosíntesis , Enterotoxinas/toxicidad , Expresión Génica/efectos de los fármacos , Bazo/metabolismo , Superantígenos/toxicidad , Linfocitos T/metabolismo , Animales , Hibridación in Situ , Interleucina-2/biosíntesis , Interleucina-2/sangre , Cinética , Lipopolisacáridos/toxicidad , Macrófagos/fisiología , Ratones , Ratones Endogámicos BALB C , ARN Mensajero/análisis , ARN Mensajero/biosíntesis , Salmonella typhimurium/inmunología , Bazo/inmunología , Staphylococcus aureus/inmunología , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunología , Transcripción Genética/efectos de los fármacos , Factor de Necrosis Tumoral alfa/análisis , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
Rabies virus (RABV) infection is characterized by the rapid neuronal spread of RABV into the CNS before a protective immune response is raised. Therefore, a typical feature of RABV infection is the paucity of inflammatory reactions in the brain. Here we examined whether the induction of immunosuppressive neuropeptides, in particular CGRP, may contribute to the ability of RABV to evade immune responses. RABV infection of mice caused a strong induction of calcitonin gene-related peptide (CGRP) in neurons and fibres in the neocortex as well as in the dentate gyrus and CA1 region of the hippocampus although RABV did not infect neurons in which CGRP expression was upregulated. Neuropeptide Y (NPY) or vasoactive intestinal peptide (VIP) expressing neurons also were not infected by RABV. In contrast, somatostatin neurons were infected by RABV. There was evidence for an RABV-induced increase of VIP and somatostatin but not of NPY. To test how CGRP expression is related to TNFalpha-induced enhancement of CNS innate and adaptive immunity during RABV infection, we used recombinant RABVs that contained either an active (SPBN-TNFalpha(+)) or an inactive (SPBN-TNFalpha(-)) TNFalpha gene. As compared to SPBN-TNFalpha(-), infection with SPBN-TNFalpha(+) attenuated the induction of CGRP but simultaneously enhanced induction of the invariant chain of MHC II, microglial activation and T cell infiltration. In conclusion, distinct neuropeptidergic neurons in the brain are remarkably spared from RABV infection suggesting a pivotal role of neuropeptides during CNS virus infection. Given the inhibitory effect of CGRP on antigen presentation, we propose that the strong RABV-induced upregulation of CGRP in the brain may contribute to the mechanism by which RABV escapes immune detection. Targeting the expression of neuropeptides, in particular CGRP, that are induced during RABV infection may open a new avenue for therapeutic intervention in human rabies.
Asunto(s)
Encéfalo/metabolismo , Enfermedades Virales del Sistema Nervioso Central/inmunología , Neuropéptidos/metabolismo , Virus de la Rabia/inmunología , Rabia/inmunología , Animales , Encéfalo/patología , Encéfalo/virología , Enfermedades Virales del Sistema Nervioso Central/metabolismo , Enfermedades Virales del Sistema Nervioso Central/patología , Femenino , Ratones , Neuropéptidos/genética , Rabia/metabolismo , Rabia/patología , Regulación hacia ArribaRESUMEN
Highly effective HIV-1-neutralizing antibodies could have utility in the prevention or treatment of HIV-1 infection. To improve the potency of 10E8, an antibody capable of near pan-HIV-1 neutralization, we engineered 10E8-surface mutants and screened for improved neutralization. Variants with the largest functional enhancements involved the addition of hydrophobic or positively charged residues, which were positioned to interact with viral membrane lipids or viral glycan-sialic acids, respectively. In both cases, the site of improvement was spatially separated from the region of antibody mediating molecular contact with the protein component of the antigen, thereby improving peripheral semi-specific interactions while maintaining unmodified dominant contacts responsible for broad recognition. The optimized 10E8 antibody, with mutations to phenylalanine and arginine, retained the extraordinary breadth of 10E8 but with â¼10-fold increased potency. We propose surface-matrix screening as a general method to improve antibodies, with improved semi-specific interactions between antibody and antigen enabling increased potency without compromising breadth.
Asunto(s)
Anticuerpos Neutralizantes/inmunología , Anticuerpos Anti-VIH/inmunología , VIH-1/inmunología , Membrana Celular/metabolismo , Proteína gp41 de Envoltorio del VIH/metabolismo , Semivida , Humanos , Pruebas de Neutralización , Polisacáridos/metabolismo , Unión ProteicaRESUMEN
BACKGROUND AND PURPOSE: Intraglandular injection of botulinum toxin (BoNT) leads to a transient denervation of the submandibular gland and this is associated with reduced salivary secretion. The purpose of the present study was to verify whether temporary acinar atrophy occurs simultaneously with chemical denervation of the glands. EXPERIMENTAL APPROACH: Tissue specimens of the right submandibular gland taken from 18 Wistar rats after intraglandular injection of BoNT A, BoNT B, or a combination of both were examined. As a sham control, an equivalent volume of saline was injected into the left submandibular gland. Morphometric measurements, immunohistochemistry, electron microscopy and western blot analysis were used to analyse the morphological and functional changes of the denervated glands. KEY RESULTS: Morphological and ultrastructural analyses of the cell organelles and secretory granula showed a clear atrophy of the acini, which was more prominent in glands injected with the combination of BoNT/A and B. Morphometric measurements of the glandular acini revealed a significant reduction of the area of the acinar cells after injection of BoNT (P=0.031). The expression of amylase was significantly reduced in BoNT treated glands. CONCLUSIONS AND IMPLICATIONS: Intraglandular application of BoNT induces structural and functional changes of the salivary glands indicated by glandular atrophy. These effects may be due to glandular denervation induced by the inhibition of the soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptors (SNAREs) involved in acetylcholine release at the neuroglandular junction and also specially inhibition of those involved in exocytosis of the granula of the acinar cells.
Asunto(s)
Toxinas Botulínicas Tipo A/toxicidad , Toxinas Botulínicas/toxicidad , Parasimpatectomía , Sistema Nervioso Parasimpático/efectos de los fármacos , Glándula Submandibular/efectos de los fármacos , Amilasas/análisis , Animales , Atrofia , Tamaño de la Célula , Masculino , Microscopía Electrónica , Tamaño de los Órganos/efectos de los fármacos , Ratas , Ratas Wistar , Glándula Submandibular/enzimología , Glándula Submandibular/inervación , Glándula Submandibular/ultraestructuraRESUMEN
The Rabies lyssavirus glycoprotein (RABV-G) is largely responsible for the neuroinvasiveness of the virus and the induction of antiviral immune responses. To study the effects of RABV-G we compared the G of the attenuated RABV variant SPBN with that of the pathogenic DOG4 strain. Infection via the olfactory route caused 100% mortality in mice with both virus variants. Of note, with the attenuated SPBN, progression of the disease was accelerated, microglia response less pronounced and IL-6 expression higher than in the presence of RABV-G from the pathogenic DOG4. However, while virus spread was less extensive, viral gene expression in individual neurons was actually higher in SPBN-infected brains without causing apoptosis of infected neurons. These differences between the two variants were not observed in infected neuronal cultures indicating that the effects of RABV-G on virus spread and viral gene expression depend on factors only present in the intact brain.
Asunto(s)
Antígenos Virales/genética , Antígenos Virales/metabolismo , Encéfalo/virología , Glicoproteínas/genética , Glicoproteínas/metabolismo , Neuronas/virología , Virus de la Rabia/aislamiento & purificación , Rabia/virología , Proteínas del Envoltorio Viral/genética , Proteínas del Envoltorio Viral/metabolismo , Carga Viral , Animales , Apoptosis , Modelos Animales de Enfermedad , Perfilación de la Expresión Génica , Genes Virales , Ratones , Análisis de Supervivencia , VirulenciaRESUMEN
Inhibitors of angiogenesis hold potential in the treatment of cancer and other diseases where the disease is caused or maintained by the inappropriate growth of blood vessels. In the present study, a novel inhibitor of angiogenesis was synthesized by covalently linking a nonanticoagulating derivative of heparin, heparin adipic hydrazide (HAH), by an acid-labile bond to the antiangiogenic steroid, cortisol. The rationale was that the heparin derivative, which binds to sulfated polyanion receptors on endothelial cells, should concentrate the steroid on the surface of vascular endothelial cells. Endocytosis of the conjugate and decomposition of the acid-labile linkage inside lysosomes and other acidic intracellular compartments should then lead to release of the cortisol and expression of its antiproliferative activity. Analysis of the stability of HAH-cortisol showed that it was stable at pH 7.4 and broke down rapidly (t1/2 15 min) at pH 4.8 at 37 degrees C. Treatment of murine pulmonary capillary endothelial cells with HAH-cortisol at 10(-5) M (with respect to cortisol) suppressed their DNA synthesis by 50% and inhibited their migration into wounded areas of confluent monolayers. HAH-cortisol at 10(-4) M (with respect to cortisol) did not suppress the DNA synthesis of Lewis lung carcinoma cells. Daily i.p. injections of HAH-cortisol into mice bearing s.c. sponge implants retarded vascularization of the sponge, and injections directly into the sponge abolished vascularization for as long as the injections were continued. Daily i.v. injections of HAH-cortisol at doses causing no apparent toxicity retarded the growth of solid s.c. Lewis lung carcinomas in mice by up to 65%. In all of these assays, equivalent treatments with a mixture of the HAH plus cortisol was significantly less effective. The antiproliferative effect of HAH-cortisol on endothelial cells appeared independent of the glucocorticoid activity of the steroid since HAH conjugated to 5 beta-pregnane-3 alpha,17 alpha,21-triol-20-one, a steroid lacking glucocorticoid or mineralocorticoid activity, was even more effective at inhibiting DNA synthesis by murine pulmonary capillary endothelial cells than was HAH-cortisol. In conclusion, HAH-cortisol represents the prototype of a new class of angiogenesis inhibitors for the treatment of cancer and other angiogenic diseases.
Asunto(s)
Heparina/análogos & derivados , Heparina/uso terapéutico , Hidrocortisona/análogos & derivados , Hidrocortisona/uso terapéutico , Neoplasias Pulmonares/irrigación sanguínea , Neovascularización Patológica/prevención & control , Animales , Anticoagulantes/uso terapéutico , Anticoagulantes/toxicidad , Antineoplásicos/uso terapéutico , Antineoplásicos/toxicidad , División Celular/efectos de los fármacos , Células Cultivadas , Cortodoxona/análogos & derivados , Cortodoxona/uso terapéutico , ADN de Neoplasias/biosíntesis , Estabilidad de Medicamentos , Endotelio Vascular/citología , Endotelio Vascular/efectos de los fármacos , Heparina/toxicidad , Hidrocortisona/toxicidad , Concentración de Iones de Hidrógeno , Neoplasias Pulmonares/tratamiento farmacológico , Neoplasias Pulmonares/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Trasplante de Neoplasias , PorcinosRESUMEN
Antidepressants produce various immunomodulatory effects, as well as an attenuation of the behavioral responses to immune challenges, such as lipopolysaccharide (LPS). To explore further the effects of antidepressants on neuroimmune interactions, rats were treated daily with either fluoxetine (Prozac) or saline for 5 weeks, and various behavioral, neuroendocrine, and immune functions were measured following administration of either LPS or saline. Chronic fluoxetine treatment significantly attenuated the anorexia and body weight loss, as well as the depletion of CRH-41 from the median eminence and the elevation in serum corticosterone levels induced by LPS. Chronic treatment with imipramine also attenuated LPS-induced adrenocortical activation. In rats and in mice, which normally display a biphasic body temperature response to LPS (initial hypothermia followed by hyperthermia), chronic treatment with fluoxetine completely abolished the hypothermic response and facilitated and strengthened the hyperthermic response. The effects of antidepressants on the responsiveness to LPS are probably not mediated by their effects on peripheral proinflammatory cytokine production, because LPS-induced expression of TNFalpha and IL-1beta mRNA in the spleen (assessed by semiquantitative in situ hybridization) was not altered following chronic treatment with either fluoxetine or imipramine. The effects of antidepressants on the acute phase response may have important clinical implications for the psychiatric and neuroendocrine disturbances that are commonly associated with various medical conditions.
Asunto(s)
Antidepresivos/farmacología , Conducta Animal/efectos de los fármacos , Encéfalo/efectos de los fármacos , Sistema Inmunológico/efectos de los fármacos , Lipopolisacáridos/farmacología , Neuroinmunomodulación/efectos de los fármacos , Glándulas Suprarrenales/efectos de los fármacos , Glándulas Suprarrenales/metabolismo , Inhibidores de Captación Adrenérgica/farmacología , Animales , Conducta Animal/fisiología , Temperatura Corporal/efectos de los fármacos , Temperatura Corporal/fisiología , Encéfalo/inmunología , Encéfalo/metabolismo , Corticosterona/metabolismo , Citocinas/genética , Esquema de Medicación/veterinaria , Interacciones Farmacológicas/fisiología , Fluoxetina/farmacología , Sistema Hipotálamo-Hipofisario/efectos de los fármacos , Sistema Hipotálamo-Hipofisario/metabolismo , Imipramina/farmacología , Sistema Inmunológico/fisiología , Masculino , Neuroinmunomodulación/fisiología , ARN Mensajero/efectos de los fármacos , ARN Mensajero/metabolismo , Ratas , Ratas Endogámicas F344 , Inhibidores Selectivos de la Recaptación de Serotonina/farmacología , Bazo/efectos de los fármacos , Bazo/metabolismoRESUMEN
The loss of spleen may lead to fatal bacterial infections. To prevent this, splenic autotransplantation has been performed in humans and experimental animals. However, there is still controversy about the protective function of this procedure. Since innervation plays an important role in splenic function, we investigated whether splenic regenerates are re-innervated, and whether this depends on the donor and host age. Splenic tissue (30 mg) was implanted into the greater omentum of either young (2 days) or old (12 months) rats, from either young or old syngeneic animals. After 3 months of regeneration, the weight of the regenerates was determined, PGP+ nerve fibers were revealed by immunohistology, and subdivided into nerve fibers of sympathetic (TH+, NPY+) or sensory (SP+, CGRP+) origin. In addition, proliferating (Ki-67 proliferation antigen+) and apoptotic cells (TUNEL technique+) were likewise investigated. No innervation of splenic regenerates was observed after implantation into old hosts, correlating with poorly developed splenic compartments. In contrast, almost normal re-innervation occurred in young hosts after implantation of both young and old splenic tissue. These regenerates showed well-developed splenic compartments and a normal number and tissue distribution of proliferating and apoptotic cells. However, after the implantation of young tissue, the final size of splenic regenerates was three times larger (140 +/- 30 vs. 40 +/- 10 mg). Thus, re-innervation of splenic implants is necessary for their subsequent development. It is determined by host age, whereas the final size of the splenic regenerates is regulated by donor age-dependent factors. This model is useful for studying both the process leading to initial innervation and the consequences of this innervation.
Asunto(s)
Envejecimiento/fisiología , Regeneración Nerviosa/fisiología , Regeneración/fisiología , Bazo/inervación , Bazo/trasplante , Animales , Apoptosis/fisiología , División Celular/fisiología , Masculino , Tamaño de los Órganos/fisiología , Ratas , Ratas Endogámicas Lew , Bazo/citología , Donantes de TejidosRESUMEN
Using Western blot analyses and a quantitative ELISA, we identified the presence and developmental accumulation of the astroglial S-100 protein(s) in rat and cat visual cortex. There is a steep rise in the S-100 content, comprising mainly S-100 beta, during the time period of highest cortical malleability in both species. A possible role of the astroglial S-100 protein(s) in experience-dependent plasticity of the visual cortex of kittens was tested by infusing antiserum against this protein during the critical period for cortical malleability. Following 1 week of monocular deprivation, the ocular dominance of single cells in the visual cortex was investigated. The vast majority of cells in the hemispheres infused with anti-S-100 serum maintained binocular responses. This finding suggests that extracellular S-100 protein is essential for ocular-dominance plasticity. Infusion of S-100 beta during the critical period of cortical malleability had no effect on deprivation-induced ocular-dominance plasticity, but interfered with the experience-dependent refinement of orientation selectivity of visual cortical neurons. It is suggested that S-100 beta may play an important role in the refinement of cortical circuitries by selectively affecting active or activated neuronal compartments. As S-100 beta is synthesized in astroglial cells, the effects on neuronal plasticity imply that glia-neuronal information transfer occurs during activity-dependent plasticity. Possible underlying mechanisms are discussed on the basis of current knowledge on the S-100 protein family, especially S-100 beta (Marshak, 1990).
Asunto(s)
Corteza Cerebral/fisiología , Neuroglía/fisiología , Plasticidad Neuronal , Neuronas/fisiología , Proteínas S100/metabolismo , Transducción de Señal , Corteza Visual/fisiología , Envejecimiento/fisiología , Animales , Gatos , Comunicación Celular , Corteza Cerebral/crecimiento & desarrollo , Corteza Cerebral/metabolismo , Dominancia Cerebral , Electrofisiología/métodos , Ratas , Visión Ocular , Corteza Visual/crecimiento & desarrollo , Corteza Visual/metabolismoRESUMEN
A partial cDNA encoding a kappa-opioid receptor was isolated and used to generate specific 35S-labeled probes to investigate the gene expression of the kappa-opioid receptor in sensory, sympathetic and spinal neurons of the rat by in situ hybridization. A subpopulation of mainly small and medium-sized neurons within dorsal root and trigeminal ganglia expressed kappa-receptor mRNA, but no signal was detectable in the superior cervical ganglion. kappa-Receptor mRNA was distributed over neurons throughout the dorsal horn and in laminae VII/VIII. Highest concentrations of positive neurons were seen in laminae I/II, dorsal lamina X and in the lateral spinal nucleus. alpha-Motoneurons and glial cells were not labeled. This distribution of kappa-receptor mRNA indicates preferential functions of kappa-receptors in sensory signalling with particular importance to nociception.
Asunto(s)
Ganglios Sensoriales/metabolismo , Neuronas/metabolismo , ARN Mensajero/metabolismo , Receptores Opioides kappa/genética , Médula Espinal/metabolismo , Animales , Secuencia de Bases , Ganglios Sensoriales/citología , Ganglios Espinales/citología , Ganglios Espinales/metabolismo , Hibridación in Situ , Masculino , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Ratas , Ratas Wistar , Médula Espinal/citología , Distribución Tisular , Ganglio del Trigémino/citología , Ganglio del Trigémino/metabolismoRESUMEN
Extracorporeal shock wave therapy (ESWT) seems to be a new therapeutic strategy for chronic pain due to tendopathies. Neurophysiological mechanisms of action for pain relief following ESWT are still unknown. The aim of this study was to investigate if the analgesic effect of ESWT is caused by modulation of the endogenous spinal opioid system. Rats were treated with two different energy flux densities (0.04 and 0.11mJ/mm(2)) and immunohistochemical analysis of met-enkephalin (MRGL) and dynorphin (Dyn) was performed at 4 or 72 h after ESWT. ESWT had no modulatory influence on the expression of the spinal opioid systems. Different energy doses or repetitive treatment did not alter MRGL or Dyn immunoreactivity in the spinal cord. Furthermore, a delayed effect of ESWT at 72 h after treatment was not detectable. We conclude from these findings that the analgesic effects of ESWT treatment are not supported by endogenous opioids.
Asunto(s)
Dinorfinas/metabolismo , Encefalina Metionina/metabolismo , Litotricia/métodos , Dolor/fisiopatología , Médula Espinal/metabolismo , Animales , Distribución de Chi-Cuadrado , Inmunohistoquímica , Ratas , Ratas Wistar , Médula Espinal/fisiopatologíaRESUMEN
We analysed the effect of intraperitoneal insufflated ozonized oxygen on the anaesthetic strength generated by tribromoethanol, ketamine/xylazine, chloral hydrate, pentobarbital, and urethane in male Wistar rats. High dosages of anaesthetic drugs normally used for deep surgical anaesthesia were injected. The ozonized oxygen gas mixture was given five times daily on five consecutive days at 0.8 mg ozone/kg body weight before anaesthesia. The reflexes were measured 15, 30, 60, 90, 120, 180, and 240 min after injection of the anaesthetic drug. The sleeping time and the loss and regain of six different reflexes on noxious and non-aversive stimuli were recorded during the 4 h of observation. O(3)/O(2)-pneumoperitoneum (O(3)/O(2)-PP) reduced the sleeping time induced by tribromoethanol and ketamine/xylazine and increased it for chloral hydrate and pentobarbital. In accordance to the changes in the duration of anaesthesia, the O(3)/O(2)-PP induced significant changes in the loss of different reflexes. Additionally, the modulatory effect of the anaesthetic drugs on splenic cytokine mRNA expression was further influenced by O(3)/O(2)-PP. Thus, the influence of an oxidative stressor on anaesthetic potency and on the resting immune system has to be taken into account for experimental designs in which surgical anaesthesia is necessary for small laboratory animals.
Asunto(s)
Anestesia/veterinaria , Anestésicos/administración & dosificación , Oxígeno/farmacología , Ozono/farmacología , Neumoperitoneo Artificial/efectos adversos , ARN Mensajero/metabolismo , Análisis de Varianza , Anestesia/métodos , Animales , Citocinas/genética , Citocinas/metabolismo , Expresión Génica/efectos de los fármacos , Hibridación in Situ , Masculino , Oxígeno/administración & dosificación , Ozono/administración & dosificación , Ratas , Ratas Wistar , Reflejo/efectos de los fármacos , Bazo/metabolismo , Factores de TiempoRESUMEN
We examined the effect of five anaesthetic drugs commonly used in laboratory animal research (tribromoethanol, ketamine/xylazine, chloral hydrate, pentobarbital, and urethane) on the expression of four pro-inflammatory cytokines. The anaesthetic agents were applied at dosages normally used for deep surgical anaesthesia. Semiquantitative image analysis of interleukin (IL)-1beta, IL-2, IL-6, and tumour necrosis factor alpha (TNFalpha) mRNA expression in the spleen of male Wistar rats 4 h after application of the anaesthetic drugs showed that these had moderate immunomodulatory effects. Ketamine/xylazine, chloral hydrate, and pentobarbital enhanced the basal expression of IL-1beta and IL-6 mRNA in rat spleen, while urethane reduced splenic IL-1beta mRNA expression. Tribromoethanol, ketamine/xylazine, and urethane reduced the basal TNFalpha mRNA levels, whereas TNFalpha mRNA expression was unaffected by chloral hydrate and by pentobarbital. The data demonstrate that these anaesthetics have slight, but significant, effects on the basal immune status of rats.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Anestésicos/farmacología , Citocinas/metabolismo , Etanol/análogos & derivados , Expresión Génica/efectos de los fármacos , ARN Mensajero/metabolismo , Adyuvantes Inmunológicos/administración & dosificación , Anestésicos/administración & dosificación , Animales , Autorradiografía , Hidrato de Cloral , Hibridación in Situ , Ketamina , Masculino , Compuestos de Mostaza Nitrogenada , Pentobarbital , Ratas , Ratas Wistar , Bazo/metabolismo , UretanoRESUMEN
BACKGROUND AND OBJECTIVE: The aim of this study was to identify the expression of cyclooxygenase-1, cyclooxygenase-2, cyclooxygenase-3, and microsomal prostaglandin E synthase-1 in young and elderly subjects. MATERIAL AND METHODS: Periodontally healthy subjects were divided into young (18-30 years, n = 7) and elderly (46-77 years, n = 7). A gingival biopsy was taken at baseline. After experimental gingivitis, clinical examination was repeated and a second biopsy was taken. The expression of cyclooxygenase-1, cyclooxygenase-2, cyclooxygenase-3, and microsomal prostaglandin E synthase-1 was analyzed by means of immunohistochemistry. RESULTS: In both healthy age groups, cyclooxygenase-1 and microsomal prostaglandin E synthase-1 were expressed in epithelial cells, endothelial cells and fibroblast-like connective tissue cells. Cyclooxygenase-1 was found in Langerhans' cells of the epithelium. Cyclooxygenase-2 expression was observed in cells exhibiting the morphology of epithelial mitosis cells, and the expression of cyclooxygenase-2 in periodontally healthy elderly subjects was significantly lower (p < or = 0.05). Following experimental gingivitis, cyclooxygenase-1 and microsomal prostaglandin E synthase-1 expression did not change. However, the expression of cyclooxygenase-2 was significantly increased in both age groups (p < or = 0.05). Cyclooxygenase-3 was not detected in any group investigated. CONCLUSION: Cyclooxygenase-1 and microsomal prostaglandin E synthase-1 were expressed constitutively in gingival tissue, and expression was unaffected by age or inflammation states. In contrast, the expression of cyclooxygenase-2 was weaker in elderly subjects. In the course of experimental gingivitis, cyclooxygenase-2 was induced in both age groups.
Asunto(s)
Encía/enzimología , Gingivitis/enzimología , Prostaglandinas/biosíntesis , Adolescente , Adulto , Factores de Edad , Anciano , Biopsia/métodos , Ciclooxigenasa 1/análisis , Ciclooxigenasa 2/análisis , Métodos Epidemiológicos , Femenino , Encía/citología , Gingivitis/etiología , Humanos , Oxidorreductasas Intramoleculares/análisis , Masculino , Persona de Mediana Edad , Prostaglandina-E Sintasas , Prostaglandina-Endoperóxido Sintasas/análisisRESUMEN
The aim of this study was to test whether repetitive pretreatments of rats with ozonized oxygen at relatively low gas volumes into the abdomen (20 ml per rat per day) have any beneficial or detrimental effects on the course of a polymicrobial-induced lethal peritonitis. Peritonitis was induced in a surgical or a nonsurgical model by usage of fecal material from the cecum. As the biological read out we used the mortality analysis. To include possible mechanisms by which ozone might influence the septic outcome, we characterized the gene expression of the pro-inflammatory cytokines IL-1beta, IL-2, and TNF-alpha mRNA in lymphoid organs. In both models, we found a significant beneficial influence of a dose-dependent O(2)/O(3 )pneumoperitoneum on the survival rate when compared to control animals or to room air. The ozone-enhanced survival seems to be independent from altered cytokine expression because there were no differences noticed in the levels of bacterial-induced gene expression of IL-1beta and TNF-alpha in septic animals pretreated with ozonized oxygen when compared to control animals.
Asunto(s)
Control de Infecciones , Oxígeno/administración & dosificación , Ozono/administración & dosificación , Peritonitis/microbiología , Neumoperitoneo Artificial , Animales , Expresión Génica , Infecciones/genética , Infecciones/mortalidad , Interleucina-1/genética , Masculino , Ratas , Ratas Wistar , Retratamiento , Factor de Necrosis Tumoral alfa/genéticaRESUMEN
To investigate the involvement of various cellular and humoral aspects of immunity in the clearance of rabies virus from the central nervous system, (CNS), we studied the development of clinical signs and virus clearance from the CNS in knockout mice lacking either B and T cells, CD8+ cytotoxic T cells, B cells, alpha/beta interferon (IFN-alpha/beta) receptors, IFN-gamma receptors, or complement components C3 and C4. Following intranasal infection with the attenuated rabies virus CVS-F3, normal adult mice of different genetic backgrounds developed a transient disease characterized by loss of body weight and appetite depression which peaked at 13 days postinfection (p.i.). While these animals had completely recovered by day 21 p.i., mice lacking either B and T cells or B cells alone developed a progressive disease and succumbed to infection. Mice lacking either CD8+ T cells, IFN receptors, or complement components C3 and C4 showed no significant differences in the development of clinical signs by comparison with intact counterparts having the same genetic background. However, while infectious virus and viral RNA could be detected in normal control mice only until day 8 p.i., in all of the gene knockout mice studied except those lacking C3 and C4, virus infection persisted through day 21 p.i. Analysis of rabies virus-specific antibody production together with histological assessment of brain inflammation in infected animals revealed that clearance of CVS-F3 by 21 days p.i. correlated with both a strong inflammatory response in the CNS early in the infection (day 8 p.i.), and the rapid (day 10 p.i.) production of significant levels of virus-neutralizing antibody (VNA). These studies confirm that rabies VNA is an absolute requirement for clearance of an established rabies virus infection. However, for the latter to occur in a timely fashion, collaboration between VNA and inflammatory mechanisms is necessary.
Asunto(s)
Anticuerpos Antivirales/inmunología , Encéfalo/inmunología , Virus de la Rabia/inmunología , Rabia/inmunología , Rabia/virología , Animales , Anticuerpos Antivirales/biosíntesis , Antígenos Virales/inmunología , Encéfalo/patología , Encéfalo/virología , Sistema Nervioso Central/inmunología , Sistema Nervioso Central/patología , Sistema Nervioso Central/virología , Modelos Animales de Enfermedad , Inflamación , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , ARN Viral/inmunología , Rabia/patología , Rabia/fisiopatologíaRESUMEN
Interleukin-12 (IL-12) is a heterodimeric cytokine that plays an important role in the regulation of the immune response. For biological activity the expression of both subunits of IL-12, p35 and p40, is required. Moreover, in the mouse the p40 chain of IL-12 specifically inhibits the effects of the IL-12 heterodimer. In the present study we have analyzed by in situ hybridization the expression of the p35 and p40 mRNA in the spleens of BALB/c and mutant (SCID, nude, beige) mice, unstimulated and after in vivo stimulation with lipopolysaccharide (LPS) and with staphylococcal enterotoxin B (SEB). In unstimulated spleens of BALB/c mice p35 and p40 mRNA were only detectable in a few strongly stained single cells, p35 mRNA was expressed in addition weakly in the B cell areas. After injection of LPS or SEB, p40 mRNA was strongly induced in the T cell areas all over the spleen, whereas expression of p35 mRNA and its distribution pattern did not change. Surprisingly, most of the mRNA for p35 and p40 was localized in different areas of the spleen and was apparently produced by different cells. In macrophage-depleted spleens the increased expression of p40 mRNA in response to LPS was reduced but still detectable, demonstrating that other cells besides macrophages can up-regulate IL-12 p40 mRNA. Nude mice showed a stronger expression of p35 mRNA, SCID mice lacked the weak p35 staining of the B cell areas but showed a strong basal expression of both p35 and p40 mRNA and a focal response to LPS. The pattern of IL-12 mRNA expression in beige mice was the same as in normal mice. These data demonstrate a spatial dissociation of expression of the two chains of IL-12 and are compatible with a regulatory role of the isolated IL-12 p40 chain in vivo. In addition, they indicate that the demonstration of mRNA for both chains of IL-12 in whole tissues or cell mixtures is not necessarily indicative of functional IL-12.
Asunto(s)
Interleucina-12/genética , Animales , Expresión Génica , Hibridación in Situ , Interleucina-12/química , Macrófagos/fisiología , Ratones , Ratones Endogámicos BALB C , Ratones Mutantes , Ratones Desnudos , Ratones SCID , ARN Mensajero/genética , ARN Mensajero/metabolismo , Bazo/metabolismoRESUMEN
T cells reach the lung via the pulmonary and bronchial arteries that supply the alveolar and bronchial regions. Although these regions are differentially affected by T cell-mediated diseases, the migration of T-cell subsets in these two regions has not been studied. Naive, memory, and effector T cells were injected into congenic rats and traced in sections of normal lung. All three T-cell subsets were found in large numbers in the alveolar region and exited again quickly. Only effector T cells accumulated in the lamina propria of the bronchi. Further, 72 h after injection 6% of the effector T cells still proliferated in the lung, whereas apoptotic effector T cells were only observed 1 h after injection (0.2%). Thus, not only effector and memory but also naive T cells continuously migrated through the lung. The preferential accumulation of effector T cells in the bronchial lamina propria may explain why some diseases preferentially affect the bronchial region.
Asunto(s)
Bronquios/citología , Memoria Inmunológica , Pulmón/citología , Linfocitos T/inmunología , Animales , Apoptosis , Bronquios/inmunología , División Celular , Movimiento Celular , Etiquetado Corte-Fin in Situ , Pulmón/inmunología , Ratas , Ratas Endogámicas , Subgrupos de Linfocitos TRESUMEN
Infection of immunocompetent adult rats with Borna disease virus (BDV) causes severe encephalitis and neural dysfunction. The expression of COX-2 and CGRP, genes previously shown to be implicated in CNS disease and peripheral inflammation, was dramatically upregulated in the cortical neurons of acutely BDV-infected rats. Neuronal COX-2 and CGRP upregulation was predominantly seen in brain areas where ED1-positive macrophages/microglia accumulated. In addition, COX-2 expression was strongly induced in brain endothelial cells and the number of COX-2 immunoreactive microglial cells was increased. In contrast, despite increased expression of viral antigens, neither COX-2 nor CGRP expression was altered in the CNS of BDV-infected rats treated with dexamethasone, or tolerant to BDV. Thus, increased CGRP and COX-2 expression in the BDV-infected brain is the result of the inflammatory response and likely to be involved in the pathogenesis of virus-induced encephalitis.