RESUMO
BACKGROUND: Asthma is a complex lung disease resulting from the interplay of genetic and environmental factors. To understand the molecular changes that occur during the development of allergic asthma without genetic and environmental confounders, an experimental model of allergic asthma in mice was used. Our goals were to (1) identify changes at the small molecule level due to allergen exposure, (2) determine perturbed pathways due to disease, and (3) determine whether small molecule changes correlate with lung function. METHODS: In this experimental model of allergic asthma, matched bronchoalveolar lavage (BAL) fluid and plasma were collected from three groups of C57BL6 mice (control vs sensitized and/or challenged with ovalbumin, n=3-5/group) 6 hour, 24 hour, and 48 hour after the last challenge. Samples were analyzed using liquid chromatography-mass spectrometry-based metabolomics. Airway hyper-responsiveness (AHR) measurements and differential cell counts were performed. RESULTS: In total, 398 and 368 dysregulated metabolites in the BAL fluid and plasma of sensitized and challenged mice were identified, respectively. These belonged to four, interconnected pathways relevant to asthma pathogenesis: sphingolipid metabolism (P=6.6×10-5 ), arginine and proline metabolism (P=1.12×10-7 ), glycerophospholipid metabolism (P=1.3×10-10 ), and the neurotrophin signaling pathway (P=7.0×10-6 ). Furthermore, within the arginine and proline metabolism pathway, a positive correlation between urea-1-carboxylate and AHR was observed in plasma metabolites, while ornithine revealed a reciprocal effect. In addition, agmatine positively correlated with lung eosinophilia. CONCLUSION: These findings point to potential targets and pathways that may be central to asthma pathogenesis and can serve as novel therapeutic targets.
Assuntos
Asma/metabolismo , Redes e Vias Metabólicas/imunologia , Animais , Arginina/metabolismo , Líquido da Lavagem Broncoalveolar , Glicerofosfolipídeos/metabolismo , Hipersensibilidade/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Fatores de Crescimento Neural/metabolismo , Prolina/metabolismo , Esfingolipídeos/metabolismoRESUMO
BACKGROUND AND PURPOSE: Montelukast and S-carbocysteine have been used in asthmatic patients as an anti-inflammatory or mucolytic agent respectively. S-carbocysteine also exhibits anti-inflammatory properties. EXPERIMENTAL APPROACH: Ovalbumin (OVA) sensitized BALB/c mice were challenged with OVA for 3 days followed by single OVA re-challenge (secondary challenge) 2 weeks later. Forty-eight hours after secondary challenge, mice were assessed for airway hyperresponsiveness (AHR) and cell composition in bronchoalveolar lavage (BAL) fluid. Suboptimal doses of 10 mg.kg(-1) of S-carbocysteine by intraperitoneal injection (ip), 20 mg.kg(-1) of montelukast by gavage, the combination of S-carbocysteine and montelukast or 3 mg.kg(-1) of dexamethasone as a control were administered from 1 day before the secondary challenge to the last experimental day. Isolated lung cells were cultured with OVA and montelukast to determine the effects on cytokine production. KEY RESULTS: Treatment with S-carbocysteine or montelukast reduced both AHR and the numbers of eosinophils in BAL fluid. Neutralizing IFN-gamma abolished the effects of S-carbocysteine on these airway responses. Combination of the two drugs showed further decreases in both AHR and eosinophils in the BAL fluid. Goblet cell metaplasia and Th2-type cytokines, interleukin (IL)-4, IL-5 and IL-13, in BAL fluid were decreased with montelukast treatment. Conversely, S-carbocysteine increased Th1-type cytokines, IFN-gamma and IL-12 in BAL fluid. CONCLUSIONS AND IMPLICATIONS: The combination of two agents, montelukast and S-carbocysteine, demonstrated additive effects on AHR and airway inflammation in a secondary allergen model most likely through independent mechanisms of action.
Assuntos
Acetatos/farmacologia , Hiper-Reatividade Brônquica/tratamento farmacológico , Carbocisteína/farmacologia , Quinolinas/farmacologia , Acetatos/administração & dosagem , Animais , Antiasmáticos/administração & dosagem , Antiasmáticos/farmacologia , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/farmacologia , Hiper-Reatividade Brônquica/imunologia , Líquido da Lavagem Broncoalveolar , Carbocisteína/administração & dosagem , Ciclopropanos , Dexametasona/farmacologia , Sinergismo Farmacológico , Quimioterapia Combinada , Eosinófilos/metabolismo , Feminino , Inflamação/tratamento farmacológico , Inflamação/imunologia , Injeções Intraperitoneais , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Quinolinas/administração & dosagem , SulfetosRESUMO
Tumor necrosis factor (TNF)-alpha is a potent cytokine with immunomodulatory, proinflammatory, and pathobiologic activities. Although TNF-alpha is thought to play a role in mediating airway inflammation and airway hyperresponsiveness (AHR), its function is not well defined. TNF-alpha-deficient mice and mice expressing TNF-alpha in their lungs because of a TNF-alpha transgene placed under the control of the surfactant protein (SP)-C promoter (SP-C/TNF-alpha-transgenic mice) were sensitized to ovalbumin (OVA) and subsequently challenged with OVA via the airways; airway function in response to inhaled methacholine was monitored. In the TNF-alpha-deficient mice, AHR was significantly increased over that in controls. In contrast, the transgenic mice failed to develop AHR. In addition, sensitized/ challenged TNF-alpha-deficient mice had significantly increased numbers of eosinophils and higher levels of interleukin (IL)-5 and IL-10 in their bronchoalveolar lavage fluid than were found for control mice. However, in SP-C/TNF-alpha-transgenic mice, both the numbers of eosinophils and levels of IL-5 and IL-10 were significantly lower than in sensitized/challenged transgene-negative mice. gammadelta T cells have been shown to be activated by TNF-alpha and to negatively regulate AHR. Depletion of gammadelta T cells in the TNF-alpha-transgenic mice in the present study increased AHR, whereas depletion of these cells had no significant effect in TNF-alpha-deficient mice. These data indicate that TNF-alpha can negatively modulate airway responsiveness, controlling airway function in allergen-induced AHR through the activation of gammadelta T cells.
Assuntos
Hiper-Reatividade Brônquica/fisiopatologia , Receptores de Antígenos de Linfócitos T gama-delta/análise , Linfócitos T/fisiologia , Fator de Necrose Tumoral alfa/fisiologia , Alérgenos/imunologia , Animais , Hiper-Reatividade Brônquica/metabolismo , Testes de Provocação Brônquica , Citocinas/metabolismo , Imunoglobulina E/análise , Pulmão/metabolismo , Pulmão/patologia , Ativação Linfocitária , Cloreto de Metacolina , Camundongos , Camundongos Transgênicos , Ovalbumina/imunologia , Proteolipídeos/genética , Surfactantes Pulmonares/genética , Linfócitos T/imunologia , Fator de Necrose Tumoral alfa/genéticaRESUMO
The closely related Th2 cytokines, IL-4 and IL-13, share many biological functions that are considered important in the development of allergic airway inflammation and airway hyperresponsiveness (AHR). The overlap of their functions results from the IL-4R alpha-chain forming an important functional signaling component of both the IL-4 and IL-13 receptors. Mutations in the C terminus region of the IL-4 protein produce IL-4 mutants that bind to the IL-4R alpha-chain with high affinity, but do not induce cellular responses. A murine IL-4 mutant (C118 deletion) protein (IL-4R antagonist) inhibited IL-4- and IL-13-induced STAT6 phosphorylation as well as IL-4- and IL-13-induced IgE production in vitro. Administration of murine IL-4R antagonist during allergen (OVA) challenge inhibited the development of allergic airway eosinophilia and AHR in mice previously sensitized with OVA. The inhibitory effect on airway eosinophilia and AHR was associated with reduced levels of IL-4, IL-5, and IL-13 in the bronchoalveolar lavage fluid as well as reduced serum levels of OVA-IGE: These observations demonstrate the therapeutic potential of IL-4 mutant protein receptor antagonists that inhibit both IL-4 and IL-13 in the treatment of allergic asthma.
Assuntos
Hiper-Reatividade Brônquica/prevenção & controle , Imunossupressores/administração & dosagem , Interleucina-13/antagonistas & inibidores , Interleucina-4/antagonistas & inibidores , Ovalbumina/imunologia , Eosinofilia Pulmonar/prevenção & controle , Receptores de Interleucina-4/antagonistas & inibidores , Proteínas Recombinantes/administração & dosagem , Animais , Especificidade de Anticorpos , Subpopulações de Linfócitos B/imunologia , Hiper-Reatividade Brônquica/imunologia , Hiper-Reatividade Brônquica/patologia , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Células Cultivadas , Citocinas/antagonistas & inibidores , Citocinas/metabolismo , Regulação para Baixo/genética , Regulação para Baixo/imunologia , Eosinófilos/imunologia , Eosinófilos/patologia , Epitélio/imunologia , Epitélio/metabolismo , Feminino , Humanos , Imunoglobulina E/biossíntese , Imunofenotipagem , Imunossupressores/síntese química , Imunossupressores/farmacologia , Injeções Intraperitoneais , Injeções Subcutâneas , Interleucina-13/farmacologia , Interleucina-4/genética , Interleucina-4/farmacologia , Contagem de Leucócitos , Pulmão/imunologia , Pulmão/patologia , Macrófagos Peritoneais/imunologia , Macrófagos Peritoneais/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Muco/metabolismo , Ovalbumina/administração & dosagem , Fosforilação , Eosinofilia Pulmonar/imunologia , Eosinofilia Pulmonar/patologia , Proteínas Recombinantes/síntese química , Proteínas Recombinantes/farmacologia , Fator de Transcrição STAT6 , Baço/citologia , Baço/imunologia , Subpopulações de Linfócitos T/imunologia , Transativadores/antagonistas & inibidores , Transativadores/metabolismoRESUMO
We compared for the first time the therapeutic potential of a specific phosphodiesterase 4 (PDE4) inhibitor, rolipram, with anti-VLA-4 and anti-IL-5 in a model of secondary allergen exposure of previously sensitized and challenged mice. To address these issues, mice were sensitized and challenged with ovalbumin (OVA) (primary challenge). Six weeks later, sensitized/challenged mice were reexposed to OVA (secondary challenge) and airway response (resistance [RL] and dynamic compliance [Cdyn]) to inhaled methacholine was monitored. After secondary OVA challenge, RL significantly increased as did the number of lung inflammatory cells and IL-4 and IL-5 production in bronchoalveolar lavage fluid (BALF). Administration of rolipram, in a dose-dependent manner, significantly prevented both changes in RL and Cdyn, as well as eosinophil, lymphocyte, and neutrophil accumulation in the BALF; IL-4 and IL-5 levels in BALF were also significantly reduced. In contrast, treatment with anti-VLA-4 and anti-IL-5 only prevented changes in RL and eosinophil numbers and IL-5 production in BALF. Further, goblet cell hyperplasia was suppressed only by treatment with rolipram. None of the treatments affected OVA-specific antibody levels. These studies confirm that IL-5 dependent eosinophilic inflammation plays an essential role in the development of certain aspects of airway function after rechallenge of sensitized mice and that lymphocytes and neutrophils are also important in the development of altered airway function. The use of agents that inhibit PDE4 may have an important role in the treatment of asthma in previously sensitized mice.
Assuntos
Alérgenos/imunologia , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Inibidores de Fosfodiesterase/farmacologia , Rolipram/farmacologia , Resistência das Vias Respiratórias/efeitos dos fármacos , Animais , Líquido da Lavagem Broncoalveolar , Feminino , Inflamação , Integrina alfa4beta1 , Integrinas/antagonistas & inibidores , Interleucina-5/antagonistas & inibidores , Complacência Pulmonar/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/farmacologia , Receptores de Retorno de Linfócitos/antagonistas & inibidoresRESUMO
The development of airway hyperresponsiveness (AHR) is correlated with the infiltration into the lungs of activated eosinophils and T lymphocytes. In large part, influx of eosinophils into the lung is dependent on very late activating antigen-4 (VLA-4) expression. However, the kinetics of eosinophil recruitment and the development of AHR are not fully delineated. Airway function was monitored by changes in lung resistance (RL) and dynamic compliance (Cdyn) to methacholine (MCh) inhalation after anti-VLA-4. After ovalbumin (OVA) sensitization and airway challenge of BALB/c mice, AHR increased as did the number of lung inflammatory cells. Administration of anti-VLA-4 to sensitized mice 2 h before the first (of three) OVA airway challenges significantly prevented changes in RL. Moreover, injection of the antibody from 2 h before the first challenge to 42 h after the last challenge significantly prevented the increases in RL, as well as eosinophil and lymphocyte numbers in the bronchoalveolar lavage fluid (BALF); interleukin-5 (IL-5) and leukotriene concentrations in BALF were also significantly inhibited. Interestingly, treatment with anti-VLA-4 only prevented changes in Cdyn and goblet cell hyperplasia when administered 2 h before the first challenge. These studies demonstrate that the timing of anti-VLA-4 administration can selectively affect pathologic processes that contribute to altered airway function in the central and peripheral airways after allergen challenge.
Assuntos
Resistência das Vias Respiratórias/imunologia , Asma/imunologia , Hiper-Reatividade Brônquica/imunologia , Integrinas/antagonistas & inibidores , Receptores de Retorno de Linfócitos/antagonistas & inibidores , Animais , Anticorpos Monoclonais/farmacologia , Asma/patologia , Brônquios/imunologia , Brônquios/patologia , Hiper-Reatividade Brônquica/patologia , Testes de Provocação Brônquica , Eosinófilos/imunologia , Eosinófilos/patologia , Feminino , Integrina alfa4beta1 , Integrinas/fisiologia , Pulmão/imunologia , Pulmão/patologia , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Receptores de Retorno de Linfócitos/fisiologiaRESUMO
In mice, respiratory syncytial virus (RSV) infection can enhance the consequences of allergic airway sensitization, resulting in lung eosinophilia and the development of airway hyperresponsiveness (AHR) to inhaled methacholine (MCh). To delineate a role for interleukin-5 (IL-5), interleukin-4 (IL-4), and interferon gamma (IFN-gamma) in mediating the effects of RSV infection on subsequent allergic sensitization, we treated BALB/c mice with anti-IL-5 during acute RSV infection but not during subsequent exposure to ovalbumin (OVA). IL-5-deficient and IL-4-deficient mice were also treated with IL-5 either during acute RSV infection or during the sensitization period. Airway responsiveness to inhaled MCh was assessed and numbers of lung eosinophils were monitored. Anti-IL-5 treatment during RSV infection reduced AHR and lung eosinophilia after subsequent exposure to allergen. In IL-5-deficient or IL-4-deficient mice lung eosinophilia and AHR after RSV infection and allergen exposure were also markedly reduced. IL-5 administration during RSV infection restored the responses to allergen in both IL-5- and IL-4-deficient mice. However, IL-5 administration only during sensitization restored these responses in IL-4-deficient but not in IL-5-deficient animals. IFN-gamma-deficient mice developed AHR and some lung eosinophilia after allergen exposure alone and when RSV infection preceded allergen, these responses were enhanced. We conclude that both IL-5, particularly during acute infection, and IL-4 are critical in mediating the effects of RSV infection on allergic airway sensitization, resulting in the development of AHR and lung eosinophilia.
Assuntos
Interleucina-4/fisiologia , Interleucina-5/fisiologia , Hipersensibilidade Respiratória/etiologia , Infecções por Vírus Respiratório Sincicial/etiologia , Animais , Feminino , Imunização , Interferon gama/fisiologia , Interleucina-4/farmacologia , Interleucina-5/farmacologia , Cloreto de Metacolina , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Eosinofilia Pulmonar/etiologia , Infecções por Vírus Respiratório Sincicial/fisiopatologiaRESUMO
Within the respiratory epithelium of asthmatic patients, copper/zinc-containing superoxide dismutase (Cu/Zn SOD) is decreased. To address the hypothesis that lung Cu/Zn SOD protects against allergen-induced injury, wild-type and transgenic mice that overexpress human Cu/Zn SOD were either passively sensitized to ovalbumin (OVA) or actively sensitized by repeated airway exposure to OVA. Controls included nonsensitized wild-type and transgenic mice given intravenous saline or airway exposure to saline. After aerosol challenge to saline or OVA, segments of tracheal smooth muscle were obtained for in vitro analysis of neural control. In response to electrical field stimulation, wild-type sensitized mice challenged with OVA had significant increases in cholinergic reactivity. Conversely, sensitized transgenic mice challenged with OVA were resistant to changes in neural control. Stimulation of tracheal smooth muscle to elicit acetylcholine release showed that passively sensitized wild-type but not transgenic mice released more acetylcholine after OVA challenge. Function of the M(2) muscarinic autoreceptor was preserved in transgenic mice. These results demonstrate that murine airways with elevated Cu/Zn SOD were resistant to allergen-induced changes in neural control.
Assuntos
Alérgenos/imunologia , Superóxido Dismutase/biossíntese , Traqueia/enzimologia , Traqueia/imunologia , Acetilcolina/metabolismo , Acetilcolina/farmacologia , Administração por Inalação , Alérgenos/administração & dosagem , Animais , Broncoconstrição/efeitos dos fármacos , Broncoconstrição/imunologia , Broncoconstrição/fisiologia , Cromatografia Líquida de Alta Pressão , Estimulação Elétrica , Eosinófilos/citologia , Humanos , Imunização , Imuno-Histoquímica , Técnicas In Vitro , Pulmão/citologia , Pulmão/imunologia , Cloreto de Metacolina/farmacologia , Camundongos , Camundongos Transgênicos , Músculo Liso/efeitos dos fármacos , Músculo Liso/metabolismo , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Receptor Muscarínico M2 , Receptores Muscarínicos/metabolismo , Traqueia/inervaçãoRESUMO
Viral respiratory infections are considered one of the triggers of exacerbations of asthma. In a model of virus-induced airway hyper-responsiveness (AHR), mice infected with human respiratory syncytial virus (RSV) were shown to develop AHR accompanied by lung eosinophilia. Inhibitors of cyclic nucleotide phosphodiesterase (PDE) have been shown to affect airway responsiveness and pulmonary allergic inflammation. In this study, we assessed the effects of type 4 PDE (PDE4) inhibitors on AHR following RSV infection and compared them with a PDE3 inhibitor. In mice infected by intranasal inoculation of RSV, treatment with the PDE4 inhibitor rolipram or Ro-20-1724 reduced both AHR and the eosinophil infiltration of the airways. In contrast, the PDE3 inhibitor, milrinone, did not influence airway responsiveness or eosinophilic inflammation. These results demonstrate that PDE4 inhibitors can modulate RSV-induced AHR and lung eosinophilia and indicate that they have a potential role in treating exacerbations of asthma triggered by viral infection.
Assuntos
3',5'-AMP Cíclico Fosfodiesterases/antagonistas & inibidores , Hiper-Reatividade Brônquica/tratamento farmacológico , Inibidores de Fosfodiesterase/farmacologia , Eosinofilia Pulmonar/tratamento farmacológico , Infecções por Vírus Respiratório Sincicial/tratamento farmacológico , Vírus Sincicial Respiratório Humano , 4-(3-Butoxi-4-metoxibenzil)-2-imidazolidinona/farmacologia , Animais , Hiper-Reatividade Brônquica/imunologia , Hiper-Reatividade Brônquica/virologia , Líquido da Lavagem Broncoalveolar , Broncoconstrição/efeitos dos fármacos , Nucleotídeo Cíclico Fosfodiesterase do Tipo 3 , Nucleotídeo Cíclico Fosfodiesterase do Tipo 4 , Citocinas/metabolismo , Eosinófilos/efeitos dos fármacos , Eosinófilos/imunologia , Feminino , Humanos , Cloreto de Metacolina , Camundongos , Camundongos Endogâmicos BALB C , Milrinona/farmacologia , Infiltração de Neutrófilos/efeitos dos fármacos , Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Eosinofilia Pulmonar/imunologia , Eosinofilia Pulmonar/virologia , Infecções por Vírus Respiratório Sincicial/complicações , Infecções por Vírus Respiratório Sincicial/imunologia , Rolipram/farmacologiaRESUMO
Cytokines play an important role in modulating inflammatory responses and, as a result, airway tone. IL-10 is a regulatory cytokine that has been suggested for treatment of asthma because of its immunosuppressive and anti-inflammatory properties. In contrast to these suggestions, we demonstrate in a model of allergic sensitization that mice deficient in IL-10 (IL-10-/-) develop a pulmonary inflammatory response but fail to exhibit airway hyperresponsiveness in both in vitro and in vivo assessments of lung function. Reconstitution of these deficient mice with the IL-10 gene fully restores development of airway hyperresponsiveness comparable to control mice. These results identify an important role of IL-10, downstream of the inflammatory cascade, in regulating the tone of the airways after allergic sensitization and challenge.
Assuntos
Hiper-Reatividade Brônquica/fisiopatologia , Interleucina-10/fisiologia , Hipersensibilidade Respiratória/fisiopatologia , Ribonucleases , Aerossóis , Animais , Proteínas Sanguíneas/análise , Testes de Provocação Brônquica , Líquido da Lavagem Broncoalveolar/química , Citocinas/análise , Estimulação Elétrica , Proteínas Granulares de Eosinófilos , Peroxidase de Eosinófilo , Eosinofilia/etiologia , Eosinofilia/fisiopatologia , Feminino , Teste de Complementação Genética , Terapia Genética , Imunização , Inflamação/fisiopatologia , Interleucina-10/deficiência , Interleucina-10/genética , Leucotrienos/análise , Pulmão/química , Pulmão/patologia , Masculino , Cloreto de Metacolina/administração & dosagem , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Músculo Liso/fisiopatologia , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Peroxidases/análise , Organismos Livres de Patógenos Específicos , Traqueia/fisiopatologiaRESUMO
The effects of an anti-CD23 monoclonal antibody (B3B4) in CD23-deficient and CD23-overexpressing mice were compared in a murine model of allergic sensitization. After sensitization and challenge with OA, mice developed increased serum levels of OA-specific IgE and IgG(1) with airway eosinophilia and AHR when compared with nonsensitized animals. Anti-CD23 treatment was studied under two protocols: 10-d OA aerosol exposure and intraperitoneal sensitization followed by aerosol challenge. In both protocols anti-CD23 significantly reduced IgE and IgG(1) levels, abolished eosinophilia, and normalized AHR in BALB/c and wild-type CD23+/+ mice but not in CD23-/- mice. These changes were associated with increases in IFN-gamma and decreases in IL-4 production, suggesting that CD23 binding may affect not only IgE production but also the Th1/Th2 imbalance during the development of allergic AHR. Absence of CD23 in gene-deficient mice significantly enhanced OA-specific IgE and IgG(1) levels, airway eosinophilia, and AHR when compared with CD23+/+ wild-type littermates after sensitization and airway challenge. Sensitized and challenged CD23 transgenic mice also developed eosinophilic airway inflammation and methacholine hyperresponsiveness. However, the extent of AHR, BAL, and tissue eosinophilia in these animals showed a significant negative correlation with levels of CD23 expression on splenic T and B cells, demonstrating a limiting role of CD23 in the development of allergic AHR.
Assuntos
Asma/imunologia , Hiper-Reatividade Brônquica/imunologia , Receptores de IgE/fisiologia , Hipersensibilidade Respiratória/imunologia , Animais , Citocinas/fisiologia , Eosinófilos/fisiologia , Feminino , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Células Th1/imunologia , Células Th2/imunologiaRESUMO
In mice, respiratory syncytial virus (RSV) infection enhances allergic airway sensitization, resulting in lung eosinophilia and in airway hyperresponsiveness (AHR). The mechanisms by which RSV contributes to development of asthma and its effects on allergic airway sensitization in mice are not known. We tested whether these consequences of RSV infection can be adoptively transferred by T cells and whether depletion of T cell subsets prevents the effects of RSV infection on subsequent airway sensitization. Mononuclear cells, T lymphocytes, or CD4 or CD8 T cells from peribronchial lymph nodes (PBLN) of RSV-infected mice were transferred into naive BALB/c mice which were then exposed to OVA via the airways. Additionally, RSV-infected mice were depleted of CD4 or CD8 T cells following acute RSV infection but prior to airway sensitization. Following sensitization, airway responsiveness to inhaled methacholine, numbers of lung eosinophils, and levels of IFN-gamma, IL-4, and IL-5 in PBLN cell cultures were monitored. Transfer of T cells from RSV-infected mice resulted in increased eosinophil influx into the lungs, increased IL-5 production, and development of AHR following airway sensitization to allergen. Transfer of CD8 but not CD4 T cells from the PBLN of RSV-infected mice also resulted in AHR following 10 days of OVA exposure. Further, depletion of CD8 T cells prevented these consequences of RSV infection while CD4 T cell depletion reduced them. We conclude that T cells, in particular CD8 T cells, are critical in mediating RSV-induced development of lung eosinophilia and AHR following allergic airway sensitization.
Assuntos
Transferência Adotiva , Hipersensibilidade Respiratória/imunologia , Infecções por Vírus Respiratório Sincicial/imunologia , Subpopulações de Linfócitos T/transplante , Subpopulações de Linfócitos T/virologia , Animais , Brônquios/citologia , Brônquios/transplante , Hiper-Reatividade Brônquica/imunologia , Hiper-Reatividade Brônquica/prevenção & controle , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Movimento Celular/imunologia , Eosinofilia/imunologia , Eosinofilia/prevenção & controle , Eosinófilos/imunologia , Eosinófilos/patologia , Feminino , Leucócitos Mononucleares/transplante , Pulmão/imunologia , Pulmão/patologia , Linfonodos/citologia , Linfonodos/transplante , Depleção Linfocítica , Camundongos , Camundongos Endogâmicos BALB C , Infecções por Vírus Respiratório Sincicial/patologia , Infecções por Vírus Respiratório Sincicial/fisiopatologiaRESUMO
While signal transducer and activator of transcription protein 6 (STAT6) is important in interleukin-4 (IL-4)-induced commitment of CD4(+) T cells to the T helper cell, type 2 (Th2) phenotype and IgE isotype switching in B cells, its role in other IL-4-mediated events and their impact upon the allergic response is less evident. In the present study we demonstrate the critical role of STAT6 in the development of allergic airway eosinophilia and airway hyperresponsiveness (AHR) after allergen sensitization and challenge. STAT6-deficient (STAT6-/-) mice did not develop a Th2 cytokine response or an allergen-specific IgE response. Further, STAT6-/- mice had a reduced constitutive and allergen-induced expression of CD23 as well as lower mucus production in the airway epithelium. Critically, we show that IL-5 alone can reconstitute airway eosinophilia and AHR in sensitized and challenged STAT6-/- mice. This emphasizes the essential nature of the IL-4-dependent signaling of T cells to the Th2 phenotype and secretion of IL-5, resulting in the airway eosinophilia and AHR. These observations underscore the importance of targeting this pathway in new antiallergic asthma drug development.
Assuntos
Hiper-Reatividade Brônquica/fisiopatologia , Eosinófilos/patologia , Interleucina-5/fisiologia , Mucosa Respiratória/patologia , Transativadores/fisiologia , Alérgenos , Animais , Animais Congênicos , Asma/fisiopatologia , Hiper-Reatividade Brônquica/imunologia , Hiper-Reatividade Brônquica/patologia , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Eosinófilos/fisiologia , Feminino , Imunização , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Inflamação/patologia , Interferon gama/análise , Interleucina-4/análise , Interleucina-5/análise , Masculino , Camundongos , Ovalbumina/imunologia , Receptores de IgE/análise , Fator de Transcrição STAT6 , Transdução de Sinais , Transativadores/deficiência , Transativadores/genéticaRESUMO
The mechanisms regulating airway function are complex and still poorly understood. In diseases such as asthma, involvement of immune-dependent mechanisms has been suggested in causing changes in airway responsiveness to bronchoconstrictors. We now demonstrate that gammadelta T cells can regulate airway function in an alphabeta T cell-independent manner, identifying them as important cells in pulmonary homeostasis. This function of gammadelta T cells differs from previously described immune-dependent mechanisms and may reflect their interaction with innate systems of host defense.
Assuntos
Receptores de Antígenos de Linfócitos T alfa-beta/imunologia , Receptores de Antígenos de Linfócitos T gama-delta/imunologia , Hipersensibilidade Respiratória/imunologia , Animais , Especificidade de Anticorpos , Líquido da Lavagem Broncoalveolar/citologia , Citocinas/análise , Pulmão/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Mutantes , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Receptores de Antígenos de Linfócitos T gama-delta/genética , Hipersensibilidade Respiratória/genéticaRESUMO
The role of IL-5 and allergen-specific IgE in the development of eosinophilic airway inflammation and airway hyperresponsiveness (AHR) was investigated in a murine model. BALB/c mice were sensitized to ovalbumin (OVA) by intraperitoneal injection on Days 1 and 14, followed by airway challenge with OVA on Days 28 and 29. Anti-IL-5 (TRFK-5) or anti-IgE (antibody 1-5) was administered before each airway challenge. Sensitized and challenged mice developed increased OVA-specific IgE serum levels, Th2 cytokine production by peribronchial lymph node (PBLN) cells, increased numbers of eosinophils (predominantly located in the peribronchial regions of the lungs), and increased airway responsiveness to methacholine (MCh). Anti-IgE treatment significantly decreased serum anti-OVA IgE levels and prevented the development of anaphylaxis but failed to affect T cell function, eosinophil airway infiltration, and AHR in sensitized and challenged mice. In contrast, treatment with anti-IL-5 antibody did not affect B cell (Ig serum levels), T cell (cytokine production), or mast cell function (immediate cutaneous reactivity) but completely inhibited development of eosinophilic lung inflammation and AHR. These data identify IL-5-mediated eosinophilia as a major target for development of AHR in this model, with little effect resulting from neutralization of IgE.
Assuntos
Hiper-Reatividade Brônquica/imunologia , Imunoglobulina E/imunologia , Interleucina-5/imunologia , Pulmão/imunologia , Análise de Variância , Animais , Anticorpos/farmacologia , Lavagem Broncoalveolar , Células Cultivadas , Citocinas/imunologia , Modelos Animais de Doenças , Ensaio de Imunoadsorção Enzimática , Eosinófilos/imunologia , Feminino , Inflamação , Pulmão/patologia , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologiaRESUMO
Allergic sensitization in asthma develops as a consequence of complex interactions between T cells and antigen-presenting cells. We have developed several in vivo models to study allergen-specific T cell and B cell function and their relevance to allergic airway hyperresponsiveness (AHR), focusing on the role of the costimulatory molecules CD80 and CD86. Treatment of mice with anti-CD86, but not anti-CD80, significantly inhibited increased serum levels of ovalbumin (OA)-specific IgE and IgG1, airway eosinophilia, and AHR both after 10 d of OA aerosol exposure (in the absence of adjuvant) and after intraperitoneal sensitization followed by repeated airway challenges. Inhibition of AHR was associated with decreased IL-4 and IL-5 levels in the BAL fluid of sensitized mice, suggesting impaired Th2 function in anti-CD86-treated animals. This effect was not seen when mice received treatment only before allergen challenge, indicating that anti-CD86 acts through inhibition of allergic sensitization and not simply by inhibiting the influx of inflammatory cells. These data suggest that the CD86 costimulatory ligand plays a major role in the development of allergic inflammation and AHR in allergen-challenged mice. Further, this study demonstrates that T-B cell interactions during allergic sensitization are amenable to therapeutic manipulation and that selective blockade of accessory signals can be an effective means for modulating distinct T cell functions.
Assuntos
Anticorpos/uso terapêutico , Antígenos CD/imunologia , Hipersensibilidade/complicações , Glicoproteínas de Membrana/imunologia , Hipersensibilidade Respiratória/tratamento farmacológico , Hipersensibilidade Respiratória/etiologia , Animais , Formação de Anticorpos/efeitos dos fármacos , Antígeno B7-2 , Líquido da Lavagem Broncoalveolar/química , Citocinas/análise , Estimulação Elétrica , Eosinofilia/tratamento farmacológico , Eosinofilia/patologia , Feminino , Imunoglobulina E/análise , Imunoglobulina G/análise , Pulmão/patologia , Camundongos , Camundongos Endogâmicos BALB C , Músculo Liso/fisiopatologia , Ovalbumina/farmacologia , Hipersensibilidade Respiratória/sangue , Traqueia/fisiopatologiaRESUMO
Viral respiratory infections can cause bronchial hyperresponsiveness and exacerbate asthma. In mice, respiratory syncytial virus (RSV) infection results in airway hyperresponsiveness (AHR) and eosinophil influx into the airways. The immune cell requirements for these responses to RSV infection are not well defined. To delineate the role of CD8 T cells in the development of RSV-induced AHR and lung eosinophilia, we tested the ability of mice depleted of CD8 T cells to develop these symptoms of RSV infection. BALB/c mice were depleted of CD8 T cells using anti-CD8 Ab treatment before intranasal administration of infectious RSV. Six days postinfection, airway responsiveness to inhaled methacholine was assessed by barometric body plethysmography, and numbers of lung eosinophils and levels of IFN-gamma, IL-4, and IL-5 in bronchoalveolar lavage fluid were monitored. RSV infection resulted in airway eosinophilia and AHR in control mice, but not in CD8-depleted animals. Further, whereas RSV-infected mice secreted increased amounts of IL-5 into the airways as compared with noninfected controls, no IL-5 was detectable in both bronchoalveolar lavage fluid and culture supernatants from CD8-depleted animals. Treatment of CD8-depleted mice with IL-5 fully restored both lung eosinophilia and AHR. We conclude that CD8 T cells are essential for the influx of eosinophils into the lung and the development of AHR in response to RSV infection.
Assuntos
Hiper-Reatividade Brônquica/imunologia , Linfócitos T CD8-Positivos/imunologia , Eosinofilia/imunologia , Infecções por Vírus Respiratório Sincicial/imunologia , Vírus Sincicial Respiratório Humano/imunologia , Doença Aguda , Animais , Hiper-Reatividade Brônquica/prevenção & controle , Linfócitos T CD8-Positivos/efeitos dos fármacos , Linfócitos T CD8-Positivos/patologia , Feminino , Humanos , Interleucina-5/metabolismo , Contagem de Linfócitos , Depleção Linfocítica , Camundongos , Camundongos Endogâmicos BALB C , Infecções por Vírus Respiratório Sincicial/complicações , Infecções por Vírus Respiratório Sincicial/patologiaRESUMO
Viral respiratory infections can cause bronchial hyperresponsiveness and exacerbate asthma. In mice, respiratory syncytial virus (RSV) infection, which induces an immune response dominated by IFN-gamma, results in airway hyperresponsiveness (AHR) and eosinophil influx into the airways, both of which are prevented by pretreatment with anti-IL-5 Ab. To delineate the role of IL-5, IL-4, and IFN-gamma in the development of RSV-induced AHR and lung eosinophilia, we tested the ability of mice deficient in each of these cytokines to develop these symptoms of RSV infection. Mice deficient in either IL-5, IL-4, or IFN-gamma were administered infectious RSV intranasally, and 6 days later, airway responsiveness to inhaled methacholine was assessed by barometric body plethysmography, and numbers of lung eosinophils and production of IFN-gamma, IL-4, and IL-5 by mononuclear cells from peribronchial lymph nodes were monitored. RSV infection resulted in airway eosinophilia and AHR in both IL-4- and IFN-gamma-deficient mice, but not in IL-5-deficient mice. Reconstitution of IL-5-deficient mice with IL-5 restored these responses and enhanced the responses in IL-4-deficient mice. Anti-VLA-4 (very late Ag-4) treatment prevented lung eosinophilia and AHR following RSV infection and IL-5 reconstitution. We conclude that in response to RSV, IL-5 is essential for the influx of eosinophils into the lung and that eosinophils in turn are critical for the development of AHR. IFN-gamma and IL-4 are not essential for these responses to RSV infection.
Assuntos
Hiper-Reatividade Brônquica/etiologia , Eosinófilos/fisiologia , Interleucina-5/fisiologia , Infecções por Vírus Respiratório Sincicial/fisiopatologia , Doença Aguda , Animais , Feminino , Integrina alfa4beta1 , Integrinas/fisiologia , Interferon gama/biossíntese , Cloreto de Metacolina/farmacologia , Camundongos , Camundongos Endogâmicos C57BL , Receptores de Retorno de Linfócitos/fisiologiaRESUMO
Aeroallergens such as Amb a I from short ragweed are important in the development of allergic airway disease. We tested the ability of a human monoclonal immunoglobulin-A (IgA) antibody specific for Amb a I (A-IgA) to modulate airway responsiveness and lung eosinophilia after airway challenge with nebulized Amb a I or ragweed extract in mice sensitized to Amb a I or ragweed extract, respectively. A-IgA or nonspecific IgA (C-IgA) were given intranasally up to 8 h before each challenge. Allergen challenge resulted in increases in airway responsiveness, in numbers of lung eosinophils, and in Amb a I-specific IgE levels. These were prevented by pretreatment with A-IgA but not with C-IgA. Decreases in IFN-gamma and increases in IL-4 and IL-5 production following challenge with Amb a I were also reduced by A-IgA treatment. In contrast, increases in total IgE and total IgG and in numbers of lung neutrophils after challenge were not significantly affected by A-IgA, which additionally induced increased levels of Amb a I-specific IgG2a antibodies. In mice sensitized and challenged with ovalbumin (OVA), A-IgA did not affect airway responsiveness, lung eosinophilia, cytokine production, or immunoglobulin levels. These data indicate that allergen-specific IgA can prevent airway hyperresponsiveness and reduce eosinophil influx into the lungs following allergen challenge via the airways in sensitized mice, and these effects are allergen-specific. Neutralization of allergen may contribute to the effects of IgA, but the induction of allergen-specific IgG2a in A-IgA-treated mice suggests an immunomodulatory action for A-IgA.
Assuntos
Hiper-Reatividade Brônquica/imunologia , Imunoglobulina A/fisiologia , Animais , Anticorpos Monoclonais , Especificidade de Anticorpos , Testes de Provocação Brônquica , Citocinas , Modelos Animais de Doenças , Eosinofilia , Feminino , Pulmão/imunologia , Camundongos , Camundongos Endogâmicos BALB CRESUMO
BACKGROUND: Systemic administration of IL-12 can prevent airway hyperresponsiveness (AHR) in mice after sensitization and repeated allergen challenge. However, systemic IL-12 has been associated with severe adverse effects. OBJECTIVE: We determined whether IL-12 administration to the airways in a dose sufficiently low so as not to result in systemic effects can modify allergic inflammation and AHR after allergen challenge. METHODS: Mice were sensitized to ovalbumin by intraperitoneal injection and challenged with ovalbumin aerosol on 3 consecutive days. During the period of challenge, IL-12 was administered intranasally following 2 regimens, designated high (1500 ng) or low (150 ng). We monitored airway responsiveness to inhaled methacholine by barometric body plethysmography, lung inflammatory cells, local cytokine production, and, to assess systemic effects of IL-12 treatment, spleen weights and numbers of eosinophils in the bone marrow. RESULTS: Allergen challenge resulted in increases in airway responsiveness and in numbers of lung eosinophils. These increases were prevented by both high- and low-dose IL-12. Additionally, IL-12 administration resulted in enhanced local interferon-gamma production and prevented the increases in local IL-4 and IL-5 production after airway challenge. A high dose, but not a low dose, of IL-12 resulted in increased spleen weights and prevented the increase in numbers of bone marrow eosinophils after allergen challenge. CONCLUSION: These data indicate that local administration of IL-12 can prevent AHR and reduce lung eosinophilia after allergen challenge in sensitized mice without eliciting systemic adverse effects. IL-12 exerts these effects by inducing local T(H1)-type responses in the airways in a setting that is normally dominated by T(H2)-type responses.