RESUMEN
Neutrophil-mediated tissue injury is a shared pathogenesis of both chronic pulmonary diseases and acute responses to pathogens, allergens, and airborne pollutants. Interventions to minimize toxic effects of neutrophil-derived oxidants and proteases are usually limited to corticosteroids, which can have adverse side effects. We used a rodent model of endotoxin-induced lung injury to test the hypothesis that the dietary supplement γ-tocopherol (γT), a natural form of vitamin E with antioxidant and novel anti-inflammatory properties, will protect from adverse nasal and pulmonary inflammatory responses induced by endotoxin (lipopolysaccharide; LPS). Male Fisher F344 rats were intranasally (i.n.) instilled with LPS for 2 consecutive days. Beginning 2 days before i.n. LPS, the rats were gavaged daily with 30mg/kg γT. Twenty-four hours after the last i.n. LPS, bronchoalveolar lavage fluid (BALF) was collected, and pulmonary and nasal tissues were analyzed for gene expression and morphometric analyses of neutrophils and intraepithelial mucosubstances (IM). LPS caused increased BALF total cells (70% increase), neutrophils (300%), protein (35%), PGE2 (500%), and secreted mucins (75%). Robust increases in neutrophils and IM were detected in conducting airways. Pulmonary expression of MUC5AC, MIP-2, CINC-1, and MCP-1 was elevated three- to eightfold by LPS. Treatment with γT inhibited LPS-induced increases in BALF total cells, neutrophils, protein, PGE2, and secreted mucins, as well as IM and tissue neutrophil influx. Furthermore γT induced the expression of the regulatory cytokines IL-10 and IFN-γ while decreasing MUC5AC, MIP-2, CINC-1, and MCP-1. These data demonstrate novel therapeutic effects of the dietary vitamin E γT promoting anti-inflammatory pathways to protect from neutrophil-mediated lung injury.
Asunto(s)
Lesión Pulmonar/tratamiento farmacológico , Mucosa Respiratoria/efectos de los fármacos , gamma-Tocoferol/administración & dosificación , Animales , Líquido del Lavado Bronquioalveolar , Suplementos Dietéticos , Regulación de la Expresión Génica/efectos de los fármacos , Lipopolisacáridos/toxicidad , Lesión Pulmonar/inducido químicamente , Lesión Pulmonar/patología , Mucinas/biosíntesis , Mucinas/genética , Mucosa Nasal/efectos de los fármacos , Mucosa Nasal/metabolismo , Neutrófilos , Ratas , Mucosa Respiratoria/metabolismoRESUMEN
PURPOSE: sPLA2-IIa is a biomarker for many inflammatory diseases in humans and is found at high levels in human tears. However, its role in ocular surface inflammation remains unclear. An experimentally induced BALB/c mouse dry eye (DE) model was used to elucidate the role of sPLA2-IIa in ocular surface inflammation. METHODS: BALB/c mice were subcutaneously injected with scopolamine and placed in a daytime air-drying device for 5 to 10 days. Control mice received no treatment. DE status was evaluated with tear production with a phenol-red thread method. Tear inflammatory cytokines were quantified by multiplex immunoassays. Ocular surface inflammation and sPLA2-IIa expression were examined by immune-staining and quantitative (q)RT(2)-PCR. Conjunctiva (CNJ) of the mice was cultured for prostaglandin E2 production induced by sPLA2-IIa with various amount of sPLA2-IIa inhibitor, S-3319. RESULTS: Treated mice produced fewer tears and heavier corneal (CN) fluorescein staining than the untreated controls (P < 0.001). They also revealed lower goblet cell density (P < 0.001) with greater inflammatory cell infiltration within the conjunctiva, and higher concentration of tear inflammatory cytokines than the controls. Moreover, treated mice showed heavier sPLA2-IIa immune staining than the controls in the CNJ epithelium, but not in the CN epithelium or the lacrimal gland. Treated mice exhibited upregulated sPLA2-IIa and cytokine gene transcription. Furthermore, CNJ cultures treated with sPLA2-IIa inhibitor showed significantly reduced sPLA2-IIa-induced inflammation. CONCLUSIONS: This is the first report regarding sPLA2-IIa in the regulation of ocular surface inflammation. The findings may therefore lead to new therapeutic strategies for ocular surface inflammation, such as DE disease.
Asunto(s)
Conjuntiva/metabolismo , Córnea/metabolismo , ADN/genética , Síndromes de Ojo Seco/genética , Regulación de la Expresión Génica , Fosfolipasas A2 Secretoras/genética , Lágrimas/metabolismo , Animales , Biomarcadores/metabolismo , Conjuntiva/patología , Córnea/patología , Modelos Animales de Enfermedad , Progresión de la Enfermedad , Síndromes de Ojo Seco/metabolismo , Síndromes de Ojo Seco/patología , Inflamación/metabolismo , Inflamación/patología , Ratones , Ratones Endogámicos BALB C , Microscopía Fluorescente , Fosfolipasas A2 Secretoras/biosíntesis , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
The mechanisms by which cannabinoid receptors CB(1) and CB(2) modulate immune function are not fully elucidated. Critical tools for the determination of the role of both receptors in the immune system are CB(1)/CB(2) double null mice (CB(1)/CB(2) null), and previous studies have shown that CB(1)/CB(2) null mice exhibit exaggerated responses to various immunological stimuli. The objective of these studies was to determine the magnitude to which CB(1)/CB(2) null mice responded to the respiratory allergen ovalbumin (OVA) as compared with wild-type C57BL/6 mice. The authors determined that in the absence of adjuvant, both wild-type and CB(1)/CB(2) null mice mounted a marked response to intranasally instilled OVA as assessed by inflammatory cell infiltrate in the bronchoalveolar lavage fluid (BALF), eosinophilia, induction of mucous cell metaplasia, and IgE production. Many of the endpoints measured in response to OVA were similar in wild-type versus CB(1)/CB(2) null mice, with exceptions being modest reductions in OVA-induced IgE and attenuation of BALF neutrophilia in CB(1)/CB(2) null mice as compared with wild-type mice. These results suggest that T-cell responses are not universally exaggerated in CB(1)/CB(2) null mice.
Asunto(s)
Alérgenos/administración & dosificación , Hiperreactividad Bronquial/inmunología , Ovalbúmina/administración & dosificación , Receptor Cannabinoide CB1/inmunología , Receptor Cannabinoide CB2/inmunología , Adyuvantes Inmunológicos , Administración Intranasal , Alérgenos/inmunología , Animales , Hiperreactividad Bronquial/genética , Hiperreactividad Bronquial/metabolismo , Líquido del Lavado Bronquioalveolar/inmunología , Citocinas/biosíntesis , Ensayo de Inmunoadsorción Enzimática , Femenino , Inmunoglobulina E/sangre , Inmunoglobulina G/sangre , Pulmón/inmunología , Pulmón/metabolismo , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ovalbúmina/inmunología , ARN Mensajero/análisis , Receptor Cannabinoide CB1/genética , Receptor Cannabinoide CB1/metabolismo , Receptor Cannabinoide CB2/genética , Receptor Cannabinoide CB2/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Ozone (O(3)), an oxidant air pollutant in photochemical smog, principally targets epithelial cells lining the respiratory tract. However, changes in gene expression have also been reported in livers of O(3)-exposed mice. The principal aim of the present study was to determine if acute exposure to environmentally relevant concentrations of O(3) could cause exacerbation of drug-induced liver injury in mice. Overdose with acetaminophen (APAP) is the most common cause of drug-induced liver injury in developed countries. In the present study, we examined the hepatic effects of acute O(3) exposure in mice pretreated with a hepatotoxic dose of APAP. C57BL/6 male mice were fasted overnight and then given APAP (300 mg/kg ip) or saline vehicle (0 mg/kg APAP). Two hours later, mice were exposed to 0, 0.25, or 0.5 ppm O(3) for 6 h and then sacrificed 9 or 32 h after APAP administration (1 or 24 h after O(3) exposure, respectively). Animals euthanized at 32 h were given 5-bromo-2-deoxyuridine 2 h before sacrifice to identify hepatocytes undergoing reparative DNA synthesis. Saline-treated mice exposed to either air or O(3) had no liver injury. All APAP-treated mice developed marked centrilobular hepatocellular necrosis that increased in severity with time after APAP exposure. O(3) exposure increased the severity of APAP-induced liver injury as indicated by an increase in necrotic hepatic tissue and plasma alanine aminotransferase activity. O(3) also caused an increase in neutrophil accumulation in livers of APAP-treated animals. APAP induced a 10-fold increase in the number of bromodeoxyuridine-labeled hepatocytes that was markedly attenuated by O(3) exposure. Gene expression analysis 9 h after APAP revealed differential expression of genes involved in inflammation, oxidative stress, and cellular regeneration in mice treated with APAP and O(3) compared to APAP or O(3) alone, providing some indications of the mechanisms behind the APAP and O(3) potentiation. These results suggest that acute exposure to near ambient concentrations of this oxidant air pollutant may exacerbate drug-induced liver injury by delaying hepatic repair.
Asunto(s)
Acetaminofén/toxicidad , Analgésicos no Narcóticos/toxicidad , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Hepatocitos/patología , Oxidantes/toxicidad , Ozono/toxicidad , Alanina Transaminasa/sangre , Animales , Bromodesoxiuridina , Líquido del Lavado Bronquioalveolar/química , Líquido del Lavado Bronquioalveolar/citología , Enfermedad Hepática Inducida por Sustancias y Drogas/sangre , Enfermedad Hepática Inducida por Sustancias y Drogas/genética , ADN/biosíntesis , Sinergismo Farmacológico , Expresión Génica/efectos de los fármacos , Perfilación de la Expresión Génica , Hepatocitos/efectos de los fármacos , Regeneración Hepática/efectos de los fármacos , Regeneración Hepática/genética , Masculino , Ratones , Ratones Endogámicos C57BL , Necrosis/inducido químicamente , Necrosis/patología , Neutrófilos/efectos de los fármacos , Estrés Oxidativo/efectos de los fármacos , Estrés Oxidativo/genética , Organismos Libres de Patógenos EspecíficosRESUMEN
BACKGROUND: Allergy to sesame seeds is an emerging food allergy of a serious nature due to a high risk of systemic anaphylaxis. Although a mouse model to study sesame anaphylaxis is desirable, currently it is not available. Here, using a transdermal exposure model system, we tested the hypothesis that sesame seed elicits IL-4-associated IgE antibody response with consequent clinical sensitization in mice. METHODS: Groups of BALB/c mice were exposed to sesame seed extract or saline or a control food (vanilla bean extract) by transdermal applications. Systemic IgE, IgG1 and IgG2a antibody responses were examined using preoptimized ELISA. Type 2 and type 1 cytokine responses were evaluated by ex vivo antigen-mediated activation of spleen cells. Clinical response to oral sesame challenge was studied. Western blot and N-terminal amino acid sequence analyses were performed to identify the sesame allergens. RESULTS: Transdermal exposure to sesame elicited robust IgE and IgG1 but very little IgG2a antibody responses. IgE response to transdermal exposure in two high-IgE responder mice strains with disparate MHC confirmed the intrinsic allergenicity of sesame seed. Transdermal sensitization was associated with activation of IL-4 but not IFN-gamma. Furthermore, oral exposure to sesame resulted in clinical signs of systemic anaphylaxis. Western blot and sequence analysis identified four allergens including Ses i 3 and the basic subunit of 11s globulins. CONCLUSION: These data argue that transdermal exposure to sesame seed can result in IL-4 activation, IgE response and clinical sensitization for systemic anaphylaxis.
Asunto(s)
Anafilaxia/inmunología , Hipersensibilidad a los Alimentos/inmunología , Globulinas/inmunología , Proteínas de Plantas/inmunología , Sesamum/efectos adversos , Administración Cutánea , Secuencia de Aminoácidos , Anafilaxia/etiología , Animales , Western Blotting , Citocinas/análisis , Globulinas/administración & dosificación , Globulinas/efectos adversos , Inmunoglobulina E/sangre , Inmunoglobulina G/sangre , Ratones , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Proteínas de Plantas/administración & dosificación , Proteínas de Plantas/efectos adversos , Subunidades de Proteína , Semillas/efectos adversos , Semillas/inmunología , Análisis de Secuencia de ADN , Sesamum/inmunología , Bazo/inmunologíaRESUMEN
Atopic disorders include a range of conditions such as allergic asthma, -rhinitis, -conjunctivitis, -dermatitis, food and drug allergies and anaphylaxis. Induction of T helper (Th)-2 immune response with consequent IgE dependent eosinophil, basophil and mast cell mediated tissue damage is the characteristic feature of allergies. The mechanism underlying this unique and long appreciated feature of allergy is being elucidated at the molecular level with advances in our knowledge of the chemokine system. Thus, chemokines that target CCR3 in concert with Th2 cytokines appear to play a pathogenic role in allergy. In contrast, chemokines that target CXCR3 in concert with Th1 cytokine appear to play a beneficial role. Accordingly, inhibiting CCR3/Th2 pathway using CCR3 antagonists is viewed as a potentially useful strategy for anti-allergy drug development. In contrast, the idea of using CXCR3 agonists to inhibiting allergic response by promoting CXCR3/Th1 pathway faces serious concerns of their potential pro-inflammatory activities in vivo. In this article we have critically evaluated the literature examining the principle and potential of this anti-allergy drug development strategy including a summary of various compounds that are under investigation.
Asunto(s)
Hipersensibilidad/tratamiento farmacológico , Receptores de Quimiocina/agonistas , Receptores de Quimiocina/antagonistas & inhibidores , Animales , Quimiocinas/metabolismo , Diseño de Fármacos , Humanos , Hipersensibilidad/inmunología , Receptores CCR3 , Receptores CXCR3 , Receptores de Quimiocina/inmunologíaRESUMEN
Chemokines belong to a large family of structurally related proteins that play a pivotal role in immune system development and deployment. While a large number of chemokines (approximately 50) and their receptors (approximately 20) have been identified from humans or mice, only a few are known in domestic veterinary species. Recent data implicate CXCL8 (old name, IL-8), CXCL10 (old name, IP-10) (both CXC chemokines) and CCL2 (old name, MCP-1) (a CC chemokine) in veterinary infections, inflammatory diseases or reproduction. There is compelling evidence for neutrophil targeting chemokines such as CXCL8, in ovine bacterial mastitis, bovine pneumonic pasturellosis and equine chronic obstructive pulmonary disease (COPD). Monocyte and lymphocyte targeting chemokines appear to play a role in caprine arthritis encephalitis (CCL2) and canine endotoxemia (CXCL10). Interestingly CCL2 is considered a missing link between hormonal and cellular control of luteolysis. On the other hand, canine cardiovascular conditions are associated with overexpression of CCL2 and CXCL8. Furthermore, a number of veterinary viral pathogens encode chemokine/chemokine receptor like molecules or chemokine binding proteins that may help viruses to evade the immune system. Here, we provide an overview of the chemokine system and critically evaluate the current literature implicating chemokines in veterinary pathophysiology. Furthermore, we highlight promising areas for further research and discuss how and why chemokine antagonists are viewed as next generation anti-inflammatory drugs for the 21st century.