RESUMO
BACKGROUND: The underlying mechanism of allergen-specific subcutaneous immunotherapy (SCIT) is not yet fully understood, but suppression of allergen-specific Th2 cells and production of allergen-specific IgG4 antibodies are two hallmarks. The impact on the innate arm of the immune system is far less clear. OBJECTIVE: The aim of this study was to investigate the effect of birch pollen (BP) SCIT on the innate immune response in a BP SCIT mouse model. METHODS: Mice with birch pollen-induced allergic airway inflammation received weekly subcutaneous immunotherapy injections with birch pollen extract (BPE) adsorbed to alum. The effect of the BP SCIT on innate cytokine levels in lung, the number and the functionality of ILC2s and the airway inflammation was determined. RESULTS: Mice with BP allergy had an increased level of the innate cytokines IL-33, IL-25, GM-CSF and IL-5+ ILC2s in the lungs. BP SCIT suppressed the number of IL-5+ ILC2s, mast cell tryptase release, Th2 cytokine production, eosinophil recruitment and peribronchial inflammatory infiltrates. In contrast, innate cytokine production and collagen deposition in the airways were not affected. CONCLUSION AND CLINICAL RELEVANCE: BP SCIT is able to suppress the adaptive and part of the innate immune response, but this is not sufficient to inhibit collagen deposition and the IL-33 expression in the airways in mice.
Assuntos
Betula/efeitos adversos , Dessensibilização Imunológica , Interleucina-33/metabolismo , Pólen/imunologia , Rinite Alérgica Sazonal/imunologia , Rinite Alérgica Sazonal/metabolismo , Células Th2/imunologia , Células Th2/metabolismo , Alérgenos/imunologia , Animais , Líquido da Lavagem Broncoalveolar/imunologia , Citocinas/metabolismo , Dessensibilização Imunológica/efeitos adversos , Dessensibilização Imunológica/métodos , Modelos Animais de Doenças , Feminino , Imunidade Inata , Pulmão/imunologia , Pulmão/metabolismo , Linfonodos/citologia , Linfonodos/imunologia , Linfonodos/metabolismo , Mastócitos/imunologia , Mastócitos/metabolismo , Camundongos , Rinite Alérgica Sazonal/diagnóstico , Rinite Alérgica Sazonal/terapiaRESUMO
Allergic asthma is characterized by persistent chronic airway inflammation, which leads to mucus hypersecretion and airway hyperresponsiveness. Nuclear receptor Nur77 plays a pivotal role in distinct immune and inflammatory cells and is expressed in eosinophils and lung epithelium. However, the role of Nur77 in allergic airway inflammation has not been studied so far. In the present study, we determined the role of Nur77 in airway inflammation using a murine model of OVA-induced allergic airway inflammation. We found that OVA-challenged Nur77 knockout (KO) mice show significantly enhanced infiltration of inflammatory cells, including eosinophils and lymphocytes, and aggravated mucus production. The infiltration of macrophages is limited in this model and was similar in wild-type and Nur77 KO mice. Higher levels of Th2 cytokines were found in bronchoalveolar lavage fluid and draining lymph node cells of Nur77-KO mice, as well as increased serum IgG1 and IgG2a levels. Knockdown of Nur77 in human lung epithelial cells resulted in a marked increase in IκBα phosphorylation, corresponding with elevated NF-κB activity, whereas Nur77 overexpression decreased NF-κB activity. Consistently, Nur77 significantly decreased mRNA levels of inflammatory cytokines and Muc5ac expression and also attenuated mucus production in lung epithelial cells. To further corroborate these findings, we searched for association of single nucleotide polymorphisms in Nur77 gene with asthma and with the severity of bronchial hyperresponsiveness. We identified three Nur77 single nucleotide polymorphisms showing association with severity of bronchial hyperresponsiveness in asthma patients. Collectively, these findings support a protective role of Nur77 in OVA-induced airway inflammation and identify Nur77 as a novel therapeutic target for airway inflammation.
Assuntos
Células Epiteliais Alveolares/metabolismo , Inflamação/metabolismo , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/metabolismo , Hipersensibilidade Respiratória/metabolismo , Alelos , Animais , Asma/genética , Asma/imunologia , Asma/metabolismo , Asma/patologia , Líquido da Lavagem Broncoalveolar/imunologia , Citocinas/metabolismo , Modelos Animais de Doenças , Ativação Enzimática , Eosinófilos/patologia , Expressão Gênica , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Inflamação/genética , Inflamação/imunologia , Inflamação/patologia , Contagem de Leucócitos , Linfonodos/imunologia , Camundongos , Camundongos Knockout , Mucina-5AC/genética , Mucina-5AC/metabolismo , NF-kappa B/metabolismo , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/deficiência , Membro 1 do Grupo A da Subfamília 4 de Receptores Nucleares/genética , Razão de Chances , Polimorfismo de Nucleotídeo Único , Hipersensibilidade Respiratória/genética , Hipersensibilidade Respiratória/imunologia , Hipersensibilidade Respiratória/patologia , Transdução de SinaisRESUMO
Allergies arise from aberrant T helper type 2 responses to allergens. Several respiratory allergens possess proteolytic activity, which has been recognized to act as an adjuvant for the development of a Th2 response. Allergen source-derived proteases can activate the protease-activated receptor-2, have specific effects on immune cells by cleaving cell membrane-bound regulatory molecules, and can disrupt tight junctions. The protease activity can induce a non-allergen-specific inflammatory response in the airways, which will set the stage for an allergen-specific Th2 response. In this review, we will discuss the evidence for the induction of oxidative stress as an underlying mechanism in Th2 sensitization to proteolytic allergens. We will discuss recent data linking the proteolytic activity of an allergen to its potential to induce oxidative stress and how this can facilitate allergic sensitization. Based on experimental data, we propose that a less proficient anti-oxidant response to allergen-induced oxidative stress contributes to the susceptibility to allergic sensitization. Besides the effect of oxidative stress on the immune response, we will also discuss how oxidative stress can increase the immunogenicity of an allergen by chemical modification.
Assuntos
Alérgenos/imunologia , Estresse Oxidativo/fisiologia , Animais , Hipersensibilidade/enzimologia , Hipersensibilidade/genética , Hipersensibilidade/imunologia , Estresse Oxidativo/genética , Espécies Reativas de Oxigênio/metabolismoRESUMO
Hexabromocyclododecane (HBCD) has been recognized as an indoor pollutant. HBCD is added as a flame retardant to many consumer products and leaches from the products into house dust. HBCD might be potentially hazardous to the airways because of inhalation of house dust. Sensitization to house dust mite (HDM) is a risk factor for the development of allergic asthma. In this study, we examined whether HBCD can affect the immune response to HDM allergens. Bone-marrow-derived dendritic cells (BMDCs) were exposed simultaneously to HBCD and HDM in vitro. HBCD enhanced oxidative stress in HDM-pulsed BMDCs, which was accompanied by a higher production of Interleukin (IL)-6 and -10. Adoptive transfer of HDM/HBCD-exposed BMDCs into naiÌve mice resulted in enhanced levels of IL-17A after inhalational challenge with HDM. Direct mucosal exposure to HBCD during HDM inhalation enhanced IL-4 or IL-17A production, depending on the HDM extract used, but did not aggravate the eosinophilic airway inflammation or airway hyper-reactivity. Our results indicate that exposure to HBCD can have a mild immune-modulating effect by enhancing the inflammatory cytokine production in response to inhaled HDM in mice.
Assuntos
Poluentes Atmosféricos/farmacologia , Alérgenos/imunologia , Antígenos de Dermatophagoides/imunologia , Asma/imunologia , Hidrocarbonetos Bromados/farmacologia , Imunidade Celular/efeitos dos fármacos , Fatores Imunológicos/farmacologia , Animais , CamundongosRESUMO
BACKGROUND: Suppression of Th2 cytokine production by allergen-specific Th2 cells is considered to be critical for the suppression of allergic symptoms by subcutaneous immunotherapy. The aim of this study was to develop a mouse model for birch pollen (BP) immunotherapy to elucidate the underlying mechanisms that contribute to the improvement of clinical symptoms. METHODS: Mice with BP-induced allergic airway inflammation received weekly subcutaneous immunotherapy (SCIT) injections with BP extract (BPE) adsorbed to alum. The effect of an increasing dose of BPE adsorbed to a fixed concentration of alum on the suppression of airway inflammation and airway hyper-responsiveness (AHR) was determined. After 2, 4, 6 or 8 immunotherapy injections, the mice were rechallenged with the same allergen and all hallmarks of allergic asthma were evaluated. RESULTS: Suppression of the immunological parameters by immunotherapy was dependent on the BPE dose. Two injections were sufficient to suppress IL-4, IL-5, IL-13, IL-10 and IFN-γ production, eosinophil recruitment and peribronchial inflammatory infiltrates. BP-specific immunoglobulins were upregulated, but this was not sufficient to reduce AHR. Eight injections were needed to suppress AHR. The gradual reduction in AHR was inversely associated with the increase of BP IgG2a. CONCLUSIONS: BP SCIT induces an early suppression of Th2-mediated eosinophilic airway inflammation, but AHR is only effectively reduced after continued SCIT conceivably by allowing IgG2a antibody titres to build up.
Assuntos
Alérgenos/imunologia , Betula/efeitos adversos , Dessensibilização Imunológica , Pólen/imunologia , Hipersensibilidade Respiratória/imunologia , Hipersensibilidade Respiratória/terapia , Alérgenos/administração & dosagem , Animais , Asma/imunologia , Citocinas/biossíntese , Modelos Animais de Doenças , Feminino , Imunização , Inflamação/imunologia , Injeções Subcutâneas , Camundongos , Mucosa Respiratória/imunologia , Mucosa Respiratória/metabolismo , Células Th2/imunologia , Células Th2/metabolismoRESUMO
RATIONALE: Polysensitization of patients who are allergic is a common feature. The underlying immunologic mechanism is not clear. The maturation status of dendritic cells (DCs) is considered to be important for priming naive T cells in the draining lymph nodes. We hypothesized that chronic airway inflammation can induce an enhanced maturation of airway DCs and facilitate subsequent priming to neoallergens. OBJECTIVES: To investigate whether chronic airway inflammation could induce an altered activation of airway DCs in mice and whether this influences the development of allergic sensitization. METHODS: Balb/c mice were repeatedly challenged with DCs to induce a chronic airway inflammation. We evaluated (1) the induction of the main characteristic features of human asthma including persistent remodeling, (2) the maturation status of airway DCs 1 month after inflammation resolved, (3) whether this influences tolerance to inhaled neoallergen, and (4) what type of T helper response would be induced by DCs. MEASUREMENTS AND MAIN RESULTS: Airway DCs displayed a mature phenotype after complete resolution of airway eosinophilia. Inhalation of a neoallergen without any adjuvant was able to induce airway inflammation in postinflammation lungs but not in control lungs. One month after inflammation, airway DCs were able to induce Th2 polarization in naive T cells consistent with the up-regulation of the Th2 skewing molecules Ym1/2 and OX-40L compared with DCs of control airways. CONCLUSIONS: This study provides evidence that sustained maturation of DCs after resolution of Th2-mediated inflammation can contribute to polysensitization.
Assuntos
Alérgenos/imunologia , Células Dendríticas/imunologia , Inflamação/imunologia , Hipersensibilidade Respiratória , Administração por Inalação , Alérgenos/administração & dosagem , Alérgenos/genética , Animais , Modelos Animais de Doenças , Humanos , Camundongos , Camundongos Endogâmicos BALB C , FenótipoRESUMO
Although dendritic cells (DCs) play an important role in sensitization to inhaled allergens, their function in ongoing T helper (Th)2 cell-mediated eosinophilic airway inflammation underlying bronchial asthma is currently unknown. Here, we show in an ovalbumin (OVA)-driven murine asthma model that airway DCs acquire a mature phenotype and interact with CD4(+) T cells within sites of peribronchial and perivascular inflammation. To study whether DCs contributed to inflammation, we depleted DCs from the airways of CD11c-diphtheria toxin (DT) receptor transgenic mice during the OVA aerosol challenge. Airway administration of DT depleted CD11c(+) DCs and alveolar macrophages and abolished the characteristic features of asthma, including eosinophilic inflammation, goblet cell hyperplasia, and bronchial hyperreactivity. In the absence of CD11c(+) cells, endogenous or adoptively transferred CD4(+) Th2 cells did not produce interleukin (IL)-4, IL-5, and IL-13 in response to OVA aerosol. In CD11c-depleted mice, eosinophilic inflammation and Th2 cytokine secretion were restored by adoptive transfer of CD11c(+) DCs, but not alveolar macrophages. These findings identify lung DCs as key proinflammatory cells that are necessary and sufficient for Th2 cell stimulation during ongoing airway inflammation.
Assuntos
Asma/imunologia , Antígeno CD11c/imunologia , Células Dendríticas/imunologia , Pulmão/imunologia , Células Th2/imunologia , Transferência Adotiva , Aerossóis/administração & dosagem , Alérgenos/administração & dosagem , Alérgenos/imunologia , Animais , Asma/induzido quimicamente , Asma/genética , Antígeno CD11c/genética , Citocinas/imunologia , Toxina Diftérica/genética , Toxina Diftérica/imunologia , Eosinófilos/imunologia , Inflamação/induzido quimicamente , Inflamação/genética , Inflamação/imunologia , Pulmão/citologia , Macrófagos Alveolares/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Transgênicos , Ovalbumina/administração & dosagem , Células Th2/transplanteRESUMO
Organophosphate flame retardants (PFRs) are commonly used as alternatives for the banned polybrominated diphenyl ethers (PBDEs) and are ubiquitously detected in indoor dust. PFRs can be potentially hazardous to respiratory health via the inhalation of house dust. Dendritic cells (DCs) are crucial in the immunological defense against pathogens in the airways. In respiratory allergy however, an aberrant immune response is induced against innocuous proteins, like house dust mite allergens. In this study, we examined whether exposure to PFRs Triphenylphosphate (TPHP) and Tris(1,3-dichloroisopropyl) phosphate (TDCIPP) affected activation/maturation of DCs at steady state and during exposure to house dust mite allergens (HDM). Bone marrow-derived dendritic cells (BMDCs) were exposed to a concentration range of each PFR (0.1-100 µM) with or without HDM in vitro to analyze the effect on the expression of major histocompatibility complex class II (MHCII), co-stimulatory molecules and cytokine production. Concentrations of TPHP and TDCIPP of ≥50 µM were cytotoxic to BMDCs. At these cytotoxic concentrations, TPHP exposure induced an activated phenotype in steady state DCs, while HDM exposed DCs acquired a tolerogenic phenotype. In contrast, TDCIPP exposure had no effect at steady state DCs but suppressed the expression of MHCII, costimulatory molecules, and the IL-6 production in HDM exposed DCs. The cytotoxic concentrations induced the anti-oxidant enzyme hemeoxigenase-1, which is a marker for oxidative stress. These results demonstrate that PFRs can be immunotoxic for DCs and suggest the necessity to evaluate the effects on the immune system on a cellular level during the risk assessment of these alternative flame retardants.
Assuntos
Retardadores de Chama/toxicidade , Éteres Difenil Halogenados/toxicidade , Organofosfatos/toxicidade , Alérgenos/efeitos adversos , Animais , Antioxidantes/química , Células da Medula Óssea/efeitos dos fármacos , Células Dendríticas/efeitos dos fármacos , Relação Dose-Resposta a Droga , Feminino , Antígenos de Histocompatibilidade Classe II/química , Inflamação , Interleucina-6/química , Camundongos , Camundongos Endogâmicos BALB C , Estresse Oxidativo , Fenótipo , Fosfatos/toxicidade , PyroglyphidaeRESUMO
Respiratory viral infections can influence the course of asthma at different time points. Severe respiratory viral infections at early age might be associated with a higher prevalence of asthma in later childhood. In established asthma, viral infections are a frequent cause of asthma exacerbation. Epidemiological and experimental animal data can illuminate the mechanisms by which viral infections can lead to sensitization to antigen and exacerbate ongoing allergic airway inflammation. In experimental rodent models of asthma, respiratory viral infection at the time of a first inhaled antigen exposure is described to induce Th2 sensitization and to enhance the allergic response to a second encounter with the same antigen. Virus infections can modulate airway dendritic cell function by up-regulation of costimulatory molecule expression, enhanced recruitment, and by inducing an inflammatory environment, all leading to an enhanced antigen presentation and possibly changing the normal tolerogenic response to inhaled antigen into an immunogenic response. In established asthma, respiratory viral infections attract several inflammatory cells, alter receptor expression on airway smooth muscle and modulate neuroimmune mechanisms, possibly leading to exacerbation of disease. Animal data suggest that the link between respiratory viral infections and increased asthma is causally related, the viral infection acting on the immune and structural cells to enhance antigen presentation and inflammatory cell recruitment.
Assuntos
Asma/etiologia , Asma/patologia , Células Dendríticas/imunologia , Infecções Respiratórias/patologia , Alérgenos , Animais , Asma/epidemiologia , Asma/virologia , Humanos , Infecções Respiratórias/virologia , Células Th2/imunologia , Viroses/imunologiaRESUMO
The indoor pollutant hexabromocyclododecane (HBCD) has been added as flame retardant to many consumer products but detaches and accumulates in house dust. Inhalation of house dust leads to exposure to house dust mite (HDM) allergens in the presence of HBCD. Activation of dendritic cells is crucial in the sensitization to HDM allergens. The current study examined whether exposure to HBCD affected activation/maturation of HDM-exposed human dendritic cells (DC). Human monocyte-derived DC (moDC) were exposed simultaneously to HDM and a concentration range of HBCD (0.1-20 µM) in vitro. HDM exposure of moDC induced expression of co-stimulatory molecule CD80 and production of pro-inflammatory cytokines interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α. However, simultaneous exposure of moDC to HBCD and HDM enhanced the expression of antigen presenting molecule HLA-DR, co-stimulatory molecule CD86 and pro-inflammatory cytokine IL-8 depending on the dose of HBCD. Our results indicate that simultaneous exposure of HDM and HBCD can enhance the antigen presentation and maturation/activation of DC.
Assuntos
Antígenos de Dermatophagoides/imunologia , Células Dendríticas/imunologia , Hidrocarbonetos Bromados/imunologia , Poluição do Ar em Ambientes Fechados/efeitos adversos , Animais , Apresentação de Antígeno , Antígeno B7-1/metabolismo , Antígeno B7-2/metabolismo , Diferenciação Celular , Células Cultivadas , Retardadores de Chama , Antígenos HLA-DR/metabolismo , Humanos , Interleucina-6/metabolismo , Interleucina-8/metabolismo , Monócitos/imunologia , Pyroglyphidae/imunologia , Fator de Necrose Tumoral alfa/metabolismoRESUMO
BACKGROUND: Respiratory viral infections can influence the course of asthma at different time points. Severe respiratory viral infections during early age are associated with a higher prevalence of asthma in later childhood. In established asthma, viral infections are a frequent cause of asthma exacerbation. OBJECTIVES: The present review focuses on epidemiological and experimental animal data that can illuminate the mechanisms by which viral infections can lead to sensitization to antigen, and exacerbate ongoing allergic airway inflammation and focuses on the role played by dendritic cells (DCs). RESULTS: In experimental rodent models of asthma, respiratory viral infection at the time of a first inhaled antigen exposure is described to induce Th2 sensitization and to enhance the allergic response to a second encounter with the same antigen. Virus infections can modulate airway dendritic cell function by upregulation of costimulatory molecule expression, enhanced recruitment, and by inducing an inflammatory environment, all leading to an enhanced antigen presentation and possibly changing the normal tolerogenic response to inhaled antigen into an immunogenic response. In established asthma, respiratory viral infections attract several inflammatory cells, alter receptor expression on airway smooth muscle and modulate neuroimmune mechanisms, possibly leading to exacerbation of disease. CONCLUSIONS: Animal data suggest that the link between respiratory viral infections and increased asthma is causally related, the viral infection acting on the immune and structural cells to enhance antigen presentation and inflammatory cell recruitment.
Assuntos
Asma/virologia , Células Dendríticas/imunologia , Infecções Respiratórias/complicações , Viroses/imunologia , Alérgenos/imunologia , Animais , Asma/epidemiologia , Asma/imunologia , Humanos , Imunização , Inflamação/virologia , Inalação , Camundongos , Modelos Imunológicos , Prevalência , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/imunologia , Células Th2/imunologia , Viroses/complicaçõesRESUMO
Mouse models of allergic asthma are increasingly used to study the immunopathology of this complex disorder. The degree and type of airway inflammation is often studied by determination of differential cell counts on cytospins of bronchoalveolar lavage fluid (BALF) cells stained with May-Grünwald Giemsa, in which the separation of eosinophils (eos) from neutrophils (neutro) and of monocytes (mono) from activated T cells can be quite problematic. In this study, we compared differential cell counts based on morphological criteria on May-Grünwald Giemsa stained cytospins with a newly developed flow cytometric method. BAL fluid cells were identified based on forward and side scatter characteristics (FSC and SSC), autofluorescence of macrophages, and simultaneous one-step staining with antibodies for T cells (CD3-Cy-Chrome), B cells (B220-Cy-Chrome), eosinophils (CCR3-PE), and dendritic cells (DCs) (MHCII-FITC, CD11c-APC). The validity of this flow cytometric determination was tested by morphological analysis of flow-sorted cellular subsets. In an animal model of ovalbumin-induced asthma, this new method correlated very well with the differential counts based on cytospins. Flow cytometric determination of the cellular composition of BAL fluid in mouse models of asthma is a rapid and easy method that can replace differential cell counts based on morphology.
Assuntos
Asma , Líquido da Lavagem Broncoalveolar/citologia , Citometria de Fluxo/métodos , Contagem de Leucócitos/métodos , Animais , Asma/imunologia , Líquido da Lavagem Broncoalveolar/imunologia , Feminino , Leucócitos/classificação , Camundongos , Camundongos Endogâmicos BALB CRESUMO
BACKGROUND: T-cells are a main target for antiinflammatory drugs in inflammatory bowel disease. As the innate immune system is also implicated in the pathogenesis of these diseases, T-cell suppressors may not only inhibit T-cell-dependent production of proinflammatory mediators but also affect innate immune cell function. Specifically, these drugs may impair innate immune cell recruitment and activation through inhibition of T-cells or act independent of T-cell modulation. We explored the extent of immune modulation by the T-cell inhibitor tacrolimus in a murine colitis model. METHODS: We assessed the effects of tacrolimus on trinitro-benzene sulphonic acid (TNBS) colitis in wildtype and Rag2-deficient mice. The severity of colitis was assessed by means of histological scores and weight loss. We further characterized the inflammation using immunohistochemistry and by analysis of isolated intestinal leukocytes at various stages of disease. RESULTS: Tacrolimus-treated wildtype mice were less sensitive to colitis and had fewer activated T-cells. Inhibition of T-cell function was associated with strongly diminished recruitment of infiltrating neutrophils in the colon at the early stages of this model. In agreement, immunohistochemistry demonstrated that tacrolimus inhibited production of the neutrophil chemoattractants CXCL1 and CXCL2. Rag2-deficient mice displayed an enhanced baseline level of lamina propria neutrophils that was moderately increased in TNBS colitis and remained unaffected by tacrolimus. CONCLUSIONS: Both the innate and the adaptive mucosal immune system contribute to TNBS colitis. Tacrolimus suppresses colitis directly through inhibition of T-cell activation and by suppression of T-cell-mediated recruitment of neutrophils.
Assuntos
Imunidade Adaptativa/efeitos dos fármacos , Colite/tratamento farmacológico , Colite/imunologia , Neutrófilos/efeitos dos fármacos , Linfócitos T/efeitos dos fármacos , Imunidade Adaptativa/imunologia , Animais , Colite/induzido quimicamente , Proteínas de Ligação a DNA/genética , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/patologia , Toxina Diftérica/toxicidade , Modelos Animais de Doenças , Progressão da Doença , Imunossupressores/farmacologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Knockout , Neutrófilos/imunologia , Neutrófilos/patologia , Índice de Gravidade de Doença , Linfócitos T/imunologia , Linfócitos T/patologia , Tacrolimo/farmacologiaRESUMO
Tertiary lymphoid organs (TLOs) are organized aggregates of B and T cells formed in postembryonic life in response to chronic immune responses to infectious agents or self-antigens. Although CD11c+ dendritic cells (DCs) are consistently found in regions of TLO, their contribution to TLO organization has not been studied in detail. We found that CD11c(hi) DCs are essential for the maintenance of inducible bronchus-associated lymphoid tissue (iBALT), a form of TLO induced in the lungs after influenza virus infection. Elimination of DCs after the virus had been cleared from the lung resulted in iBALT disintegration and reduction in germinal center (GC) reactions, which led to significantly reduced numbers of class-switched plasma cells in the lung and bone marrow and reduction in protective antiviral serum immunoglobulins. Mechanistically, DCs isolated from the lungs of mice with iBALT no longer presented viral antigens to T cells but were a source of lymphotoxin (LT) beta and homeostatic chemokines (CXCL-12 and -13 and CCL-19 and -21) known to contribute to TLO organization. Like depletion of DCs, blockade of LTbeta receptor signaling after virus clearance led to disintegration of iBALT and GC reactions. Together, our data reveal a previously unappreciated function of lung DCs in iBALT homeostasis and humoral immunity to influenza virus.
Assuntos
Células Dendríticas/imunologia , Pulmão/imunologia , Pulmão/virologia , Tecido Linfoide/imunologia , Tecido Linfoide/virologia , Infecções por Orthomyxoviridae/imunologia , Orthomyxoviridae/imunologia , Transferência Adotiva , Animais , Formação de Anticorpos/imunologia , Apresentação de Antígeno/imunologia , Antígenos Virais/imunologia , Antígeno CD11c/imunologia , Movimento Celular , Quimiocinas/imunologia , Células Dendríticas/citologia , Células Dendríticas/virologia , Fator de Crescimento Semelhante a EGF de Ligação à Heparina , Homeostase , Peptídeos e Proteínas de Sinalização Intercelular/imunologia , Pulmão/patologia , Camundongos , Camundongos Endogâmicos C57BL , Modelos Animais , Infecções por Orthomyxoviridae/virologiaRESUMO
The World Health Organization estimates that lower respiratory tract infections (excluding tuberculosis) account for approximately 35% of all deaths caused by infectious diseases. In many cases, the cause of death may be caused by multiple pathogens, e.g., the life-threatening bacterial pneumonia observed in patients infected with influenza virus. The ability to evolve more efficient immunity on each successive encounter with antigen is the hallmark of the adaptive immune response. However, in the absence of cross-reactive T and B cell epitopes, one lung infection can modify immunity and pathology to the next for extended periods of time. We now report for the first time that this phenomenon is mediated by a sustained desensitization of lung sentinel cells to Toll-like receptor (TLR) ligands; this is an effect that lasts for several months after resolution of influenza or respiratory syncytial virus infection and is associated with reduced chemokine production and NF-kappaB activation in alveolar macrophages. Although such desensitization may be beneficial in alleviating overall immunopathology, the reduced neutrophil recruitment correlates with heightened bacterial load during secondary respiratory infection. Our data therefore suggests that post-viral desensitization to TLR signals may be one possible contributor to the common secondary bacterial pneumonia associated with pandemic and seasonal influenza infection.
Assuntos
Infecções Bacterianas/imunologia , Infecções por Orthomyxoviridae/imunologia , Infecções Respiratórias/imunologia , Receptores Toll-Like/imunologia , Animais , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Citocinas/imunologia , Citocinas/farmacologia , Células Epiteliais/imunologia , Feminino , Flagelina/farmacologia , Ligantes , Lipopolissacarídeos/farmacologia , Macrófagos Alveolares/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , NF-kappa B/imunologia , Neutrófilos/imunologia , Pseudomonas aeruginosa , Sistema Respiratório/imunologia , Streptococcus pneumoniaeRESUMO
Although dendritic cells (DCs) play an important role in mediating protection against influenza virus, the precise role of lung DC subsets, such as CD11b- and CD11b+ conventional DCs or plasmacytoid DCs (pDCs), in different lung compartments is currently unknown. Early after intranasal infection, tracheal CD11b-CD11chi DCs migrated to the mediastinal lymph nodes (MLNs), acquiring co-stimulatory molecules in the process. This emigration from the lung was followed by an accumulation of CD11b+CD11chi DCs in the trachea and lung interstitium. In the MLNs, the CD11b+ DCs contained abundant viral nucleoprotein (NP), but these cells failed to present antigen to CD4 or CD8 T cells, whereas resident CD11b-CD8+ DCs presented to CD8 cells, and migratory CD11b-CD8- DCs presented to CD4 and CD8 T cells. When lung CD11chi DCs and macrophages or langerin+CD11b-CD11chi DCs were depleted using either CD11c-diphtheria toxin receptor (DTR) or langerin-DTR mice, the development of virus-specific CD8+ T cells was severely delayed, which correlated with increased clinical severity and a delayed viral clearance. 120G8+ CD11cint pDCs also accumulated in the lung and LNs carrying viral NP, but in their absence, there was no effect on viral clearance or clinical severity. Rather, in pDC-depleted mice, there was a reduction in antiviral antibody production after lung clearance of the virus. This suggests that multiple DCs are endowed with different tasks in mediating protection against influenza virus.
Assuntos
Antígenos de Superfície/imunologia , Antígeno CD11b/imunologia , Células Dendríticas/imunologia , Lectinas Tipo C/imunologia , Pulmão/imunologia , Lectinas de Ligação a Manose/imunologia , Infecções por Orthomyxoviridae/imunologia , Orthomyxoviridae/imunologia , Animais , Anticorpos Antivirais/imunologia , Formação de Anticorpos/genética , Apresentação de Antígeno/genética , Antígenos de Superfície/genética , Antígeno CD11b/genética , Antígeno CD11c/genética , Antígeno CD11c/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/virologia , Antígenos CD8/genética , Antígenos CD8/imunologia , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/virologia , Movimento Celular/imunologia , Células Dendríticas/virologia , Cães , Feminino , Fator de Crescimento Semelhante a EGF de Ligação à Heparina , Peptídeos e Proteínas de Sinalização Intercelular/genética , Peptídeos e Proteínas de Sinalização Intercelular/imunologia , Lectinas Tipo C/genética , Pulmão/virologia , Linfonodos/imunologia , Linfonodos/virologia , Masculino , Lectinas de Ligação a Manose/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Transgênicos , Proteínas do Nucleocapsídeo/imunologia , Infecções por Orthomyxoviridae/genética , Plasmócitos/imunologia , Plasmócitos/virologia , Traqueia/imunologia , Traqueia/virologiaRESUMO
Respiratory syncytial virus (RSV) is the primary cause of bronchiolitis in young children. Upon infection both T helper 1 (Th1) and Th2 cytokines are produced. Because RSV-induced Th2 responses have been associated with severe immunopathology and aggravation of allergic reactions, the regulation of the immune response following RSV infection is crucial. In this study we examined the influence of RSV on the activation and function of murine bone marrow-derived dendritic cells (DCs). RSV induced the expression of maturation markers on myeloid DCs (mDCs) in vitro. The mDCs stimulated with RSV and ovalbumin (OVA) enhanced proliferation of OVA-specific T cells, which produced both Th1 and Th2 cytokines. In contrast to mDCs, RSV did not induce the expression of maturation markers on plasmacytoid DCs (pDCs), not did it enhance the proliferation of OVA-specific T cells that were cocultured with pDCs. However, RSV stimulated the production of interferon-alpha (IFN-alpha) by pDCs. Our findings indicate a clear difference in the functional activation of DC subsets. RSV-stimulated mDCs may have immunostimulatory effects on both Th1 and Th2 responses, while RSV-stimulated pDCs have direct antiviral activity through the release of IFN-alpha.
Assuntos
Células Dendríticas/imunologia , Vírus Sinciciais Respiratórios/imunologia , Alérgenos/imunologia , Animais , Células da Medula Óssea/imunologia , Diferenciação Celular/imunologia , Proliferação de Células , Células Cultivadas , Técnicas de Cocultura , Citocinas/biossíntese , Feminino , Interferon-alfa/biossíntese , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Células Th1/imunologia , Células Th2/imunologiaRESUMO
BACKGROUND: In allergic rhinitis (AR) CD4(+) T(H)2 lymphocytes control inflammation by secreting T(H)2 cytokines, but little is known about how these cells are activated to cause disease. OBJECTIVE: We sought to study the contribution of antigen-presenting dendritic cells (DCs) in activating T(H)2 cells and controlling allergic inflammation. METHODS: Nasal mucosal biopsy specimens were taken from patients with house dust mite allergy and perennial AR and healthy control subjects. DC numbers were evaluated by using immunohistochemistry. The functional role of DCs was studied in a novel mouse model for AR using BALB/c mice and CD11c-diphtheria toxin (DT) receptor transgenic mice. RESULTS: In symptomatic patients with perennial AR, the number of CD1a(+) and CD11c(+) MHCII(+) DCs was higher in the epithelium and lamina propria of the nasal mucosa compared with that seen in healthy control subjects. In patients with AR, DCs had a more mature (CD86(+)) phenotype and were found in close approximation with T lymphocytes. Similarly, in a mouse model of ovalbumin (OVA)-induced AR, CD11c(+) DCs accumulated in areas of nasal eosinophilic inflammation and clustered with CD4(+) T lymphocytes. CD11c(+) DCs were conditionally depleted during allergen challenge by means of systemic administration of DT to CD11c-diphtheria toxin receptor transgenic mice to address the functional role of DCs in maintaining inflammation. In the absence of CD11c(+) DCs, nasal OVA challenge in OVA-sensitized mice did not induce nasal eosinophilia and did not boost OVA-specific IgE levels or T(H)2 cytokine production in the cervical lymph nodes. Conversely, when OVA-pulsed DCs were administered intranasally to sensitized mice, they strongly enhanced OVA-induced nasal eosinophilia and T(H)2 cytokine production. CONCLUSIONS: These data in human subjects and mice suggest an essential role for nasal DCs in activation of effector T(H)2 function leading to AR. CLINICAL IMPLICATIONS: Nasal DCs play an essential role in AR and therefore constitute a novel target for therapeutic intervention.
Assuntos
Células Dendríticas/imunologia , Rinite Alérgica Perene/imunologia , Animais , Contagem de Células , Diferenciação Celular/imunologia , Células Dendríticas/metabolismo , Células Dendríticas/patologia , Modelos Animais de Doenças , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Mucosa Nasal/imunologia , Mucosa Nasal/metabolismo , Mucosa Nasal/patologia , Pyroglyphidae/imunologia , Rinite Alérgica Perene/patologia , Rinite Alérgica Sazonal/imunologia , Rinite Alérgica Sazonal/patologia , Células Th2/imunologia , Células Th2/patologiaRESUMO
Asthma is characterized by infiltration of the airway wall with eosinophils. Although eosinophils are considered to be effector cells, recent studies have reported their ability to activate primed Th2 cells. In this study, we investigated whether eosinophils are capable of presenting Ag to unprimed T cells in draining lymph nodes (DLN) of the lung and compared this capacity with professional dendritic cells (DC). During development of eosinophilic airway inflammation in OVA-sensitized and challenged mice, CCR3(+) eosinophils accumulated in the DLN. To study their function, eosinophils were isolated from the bronchoalveolar lavage fluid of mice by sorting on CCR3(+)B220(-)CD3(-)CD11c(dim) low autofluorescent cells, avoiding contamination with other APCs, and were intratracheally injected into mice that previously received CFSE-labeled OVA TCR-transgenic T cells. Eosinophils did not induce divisions of T cells in the DLN, whereas DC induced on average 3.7 divisions in 45.7% of T cells. To circumvent the need for Ag processing or migration in vivo, eosinophils were pulsed with OVA peptide and were still not able to induce T cell priming in vitro, whereas DC induced vigorous proliferation. This lack of Ag-presenting ability was explained by the very weak expression of MHC class II on fresh eosinophils, despite expression of the costimulatory molecules CD80 and ICAM-1. This investigation does not support any role for airway eosinophils as APCs to naive T cells, despite their migration to the DLN at times of allergen exposure. DC are clearly superior in activating T cells in the DLN of the lung.
Assuntos
Células Apresentadoras de Antígenos/imunologia , Movimento Celular/imunologia , Eosinófilos/imunologia , Pulmão/citologia , Pulmão/imunologia , Linfonodos/citologia , Linfonodos/imunologia , Subpopulações de Linfócitos T/imunologia , Animais , Apresentação de Antígeno , Células Apresentadoras de Antígenos/metabolismo , Líquido da Lavagem Broncoalveolar/citologia , Líquido da Lavagem Broncoalveolar/imunologia , Agregação Celular/imunologia , Células Cultivadas , Células Dendríticas/imunologia , Células Dendríticas/transplante , Eosinófilos/metabolismo , Eosinófilos/transplante , Feminino , Fator Estimulador de Colônias de Granulócitos e Macrófagos/farmacologia , Injeções Intravenosas , Interfase/imunologia , Intubação Intratraqueal , Mediastino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Transgênicos , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Receptores de Antígenos de Linfócitos T/genética , Subpopulações de Linfócitos T/metabolismoRESUMO
BACKGROUND: Airway dendritic cells (DCs) are crucial for the generation of TH2 cells from naive T cells during sensitization and for reactivation of primed TH2 cells on allergen challenge in mouse models of asthma. It is unknown whether CD80/CD86 costimulation is necessary during both phases of the response because primed T cells rely less on costimulatory molecules compared with naive T cells. OBJECTIVE: We sought to study the contribution of CD80/CD86 costimulatory molecules on DCs during sensitization or challenge in a mouse model of asthma. METHODS: Naive BALB/c mice received an intratracheal injection of ovalbumin (OVA)-pulsed DCs obtained from the bone marrow of wild-type (WT) or CD80/CD86-/- mice and were subsequently challenged with OVA aerosol to address the role of costimulation during sensitization. OVA-sensitized mice received OVA-pulsed WT or CD80/CD86-/- DCs without OVA aerosol to address the role of costimulation during challenge. RESULTS: WT DCs induced the proliferation and effector TH2 differentiation of naive OVA-specific T cells, whereas CD80/CD86-/- DCs induced only proliferation. Not surprisingly, WT DCs but not CD80/CD86-/- DCs induced sensitization to OVA in naive mice. In contrast, in OVA-sensitized mice intratracheal injection of CD80/CD86-/- OVA-pulsed DCs led to eosinophilic airway inflammation, goblet cell hyperplasia, and effector TH2 cytokine production that was not different from that seen after injection with WT OVA-DCs, even when the inducible costimulator ICOS was blocked or cytotoxic T lymphocyte-associated antigen 4 immunoglobulin was given. CONCLUSION: CD80/CD86 costimulation on DCs is only necessary during priming of naive T cells into TH2 cells but not during restimulation of previously primed TH2 cells in the challenge phase.