RESUMO
Exposure to dogs in early infancy has been shown to reduce the risk of childhood allergic disease development, and dog ownership is associated with a distinct house dust microbial exposure. Here, we demonstrate, using murine models, that exposure of mice to dog-associated house dust protects against ovalbumin or cockroach allergen-mediated airway pathology. Protected animals exhibited significant reduction in the total number of airway T cells, down-regulation of Th2-related airway responses, as well as mucin secretion. Following dog-associated dust exposure, the cecal microbiome of protected animals was extensively restructured with significant enrichment of, amongst others, Lactobacillus johnsonii. Supplementation of wild-type animals with L. johnsonii protected them against both airway allergen challenge or infection with respiratory syncytial virus. L. johnsonii-mediated protection was associated with significant reductions in the total number and proportion of activated CD11c(+)/CD11b(+) and CD11c(+)/CD8(+) cells, as well as significantly reduced airway Th2 cytokine expression. Our results reveal that exposure to dog-associated household dust results in protection against airway allergen challenge and a distinct gastrointestinal microbiome composition. Moreover, the study identifies L. johnsonii as a pivotal species within the gastrointestinal tract capable of influencing adaptive immunity at remote mucosal surfaces in a manner that is protective against a variety of respiratory insults.
Assuntos
Hiper-Reatividade Brônquica/imunologia , Poeira/imunologia , Exposição Ambiental , Trato Gastrointestinal/microbiologia , Lactobacillus/crescimento & desenvolvimento , Microbiota/genética , Infecções por Vírus Respiratório Sincicial/imunologia , Animais , Hiper-Reatividade Brônquica/prevenção & controle , Cães , Citometria de Fluxo , Fluorescência , Lactobacillus/imunologia , Pulmão/patologia , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologia , Infecções por Vírus Respiratório Sincicial/prevenção & controle , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células Th2/imunologiaRESUMO
IL-27 is a heterodimeric cytokine composed of the subunits p28 and Epstein-Barr virus induced gene (EBI)-3 and is known for its effects on T-cell function and differentiation. IL-27 signals through the widely expressed IL-27 receptor (IL-27R), composed of the ligand-specific IL-27Rα chain and gp130. Engagement of the IL-27R activates STAT1 signaling, induces the expression of the type 1 helper T-cell (Th1) cytokine, interferon γ, and suppresses the differentiation of Th2 and Th17 cells. This study investigates the role of IL-27 signaling in respiratory syncytial virus (RSV) infection using IL-27Rα-deficient mice (IL-27rKO). Analysis of lungs from RSV-infected IL-27rKO mice showed exacerbation of mucus secretion compared with wild type, as well as enhanced expression of Muc5ac and Gob5 mRNA, markers of goblet cell metaplasia/hyperplasia. When compared with wild-type mice, RSV-challenged IL-27rKO mice had enhanced expression of Th17-associated cytokine IL-17a and an imbalance between Th1 and Th2 cytokine levels. Neutralization of IL-17 in RSV-infected IL-27rKO mice resulted in a significant decrease in the pulmonary mucus response and inhibition of the Th2-associated cytokines. Interestingly, IL-17 blockage led to an increase in the expression of IL-27 subunits p28 and EBI-3 in the lungs and lymph nodes of RSV-infected mice. Thus, IL-27 functions as a regulatory cytokine during RSV pathogenesis by suppressing the development of Th17 cells, but it also appears to be regulated by IL-17 induced by the virus.
Assuntos
Interleucina-17/imunologia , Receptores de Citocinas/imunologia , Infecções por Vírus Respiratório Sincicial/imunologia , Vírus Sinciciais Respiratórios/imunologia , Células Th17/imunologia , Células Th2/imunologia , Animais , Canais de Cloreto/genética , Canais de Cloreto/imunologia , Interleucina-17/genética , Interleucinas/genética , Interleucinas/imunologia , Camundongos , Camundongos Knockout , Antígenos de Histocompatibilidade Menor , Mucina-5AC/genética , Mucina-5AC/imunologia , Mucoproteínas/genética , Mucoproteínas/imunologia , Muco/imunologia , Receptores de Citocinas/genética , Receptores de Interleucina , Infecções por Vírus Respiratório Sincicial/genética , Infecções por Vírus Respiratório Sincicial/patologia , Fator de Transcrição STAT1/genética , Fator de Transcrição STAT1/imunologia , Células Th17/patologia , Células Th2/patologiaRESUMO
Recent work demonstrated the importance of macroautophagy in dendritic cell (DC) maturation and innate cytokine production upon viral infection through delivery of cytoplasmic viral components to intracellular TLRs. To study the functional consequences of impaired autophagosome formation during a respiratory syncytial virus (RSV) infection, mice harboring significant autophagy defects due to Beclin-1 haploinsufficiency (Beclin-1(+/-)) were used. Upon RSV infection in vivo, lungs of Beclin-1(+/-) mice showed increased Th2 cytokine production, mucus secretion, and lung infiltration of eosinophils and inflammatory DCs. Although isolated airway epithelial cells from Beclin-1(+/-) mice demonstrated little change compared with wild-type mice, Beclin-1(+/-) pulmonary and bone marrow-derived DCs showed decreased expression of MHC class II and innate cytokine production upon RSV infection. Further examination indicated that Beclin-1(+/-) DCs stimulated less IFN-γ and IL-17 production by cocultured CD4(+) T cells and increased Th2 cytokine production in comparison with wild-type controls. Finally, adoptive transfer of RSV-infected Beclin-1(+/-) DCs into the airways of wild-type mice produced severe lung pathology and increased Th2 cytokine production upon subsequent RSV challenge compared with wild-type DC transfer controls. These results indicate a critical role for autophagy in DCs during pulmonary viral infection, facilitating appropriate antiviral adaptive immune responses.
Assuntos
Proteínas Reguladoras de Apoptose/imunologia , Autofagia/imunologia , Linfócitos T CD4-Positivos/imunologia , Células Dendríticas/imunologia , Infecções por Vírus Respiratório Sincicial/imunologia , Animais , Proteínas Reguladoras de Apoptose/metabolismo , Proteína Beclina-1 , Células Dendríticas/metabolismo , Feminino , Citometria de Fluxo , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Microscopia Confocal , Infecções por Vírus Respiratório Sincicial/metabolismo , Infecções por Vírus Respiratório Sincicial/patologia , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Activation of the canonical Notch pathways has been implicated in Th cell differentiation, but the role of specific Notch ligands in Th2-mediated allergic airway responses has not been completely elucidated. In this study, we show that delta-like ligand 4 (Dll4) was upregulated on dendritic cells in response to cockroach allergen. Blocking Dll4 in vivo during either the primary or secondary response enhanced allergen-induced pathogenic consequences including airway hyperresponsiveness and mucus production via increased Th2 cytokines. In vitro assays demonstrated that Dll4 regulates IL-2 in T cells from established Th2 responses as well as during primary stimulation. Notably, Dll4 blockade during the primary, but not the secondary, response increased IL-2 levels in lung and lymph node of allergic mice. The in vivo neutralization of Dll4 was associated with increased expansion and decreased apoptosis during the primary allergen sensitization. Moreover, Dll4-mediated Notch activation of T cells during primary stimulation in vitro increased apoptosis during the contraction/resting phase of the response, which could be rescued by exogenous IL-2. Consistent with the role for Dll4-mediated IL-2 regulation in overall T cell function, the frequency of IL-4-producing cells was also significantly altered by Dll4 both in vivo and in vitro. These data demonstrate a regulatory role of Dll4 both in initial Th2 differentiation and in Th2 cytokine production in established allergic responses.
Assuntos
Interleucina-2/biossíntese , Peptídeos e Proteínas de Sinalização Intracelular/imunologia , Proteínas de Membrana/imunologia , Hipersensibilidade Respiratória/imunologia , Células Th2/imunologia , Proteínas Adaptadoras de Transdução de Sinal , Animais , Proteínas de Ligação ao Cálcio , Diferenciação Celular/imunologia , Separação Celular , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Citometria de Fluxo , Interleucina-2/imunologia , Peptídeos e Proteínas de Sinalização Intracelular/metabolismo , Ativação Linfocitária/imunologia , Proteínas de Membrana/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Hipersensibilidade Respiratória/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Células Th2/citologia , Células Th2/metabolismoRESUMO
In this study, the early innate cytokine and chemokine response of murine dendritic cells (DCs) and macrophages to Mycobacterium tuberculosis infection was compared. The findings indicate a dissimilar gene expression pattern between the two cell types. The expression of IL-12 and IL-23, important for promoting Th1 and Th17 cells, respectively, was up-regulated only in DCs. In addition, expression of CCL1 and CCL17, which are important in recruitment of T regulatory cells, was DC-specific, as was the expression of the immunosuppressive cytokine IL-10. Macrophages, in contrast, exhibited enhanced expression for CCL2 and CXCL10, chemokines that recruit cells to sites of inflammation, and for mycobactericidal molecules NO synthase 2 and TNF. Together, the findings suggest that a component of the innate DC response is not only programmed toward Th1 priming but is also for controlling the magnitude of the Th1 response, and part of the macrophage response is intended for recruiting cells to the lung and for mycobactericidal functions.
Assuntos
Quimiocinas/genética , Citocinas/genética , Células Dendríticas/imunologia , Macrófagos/imunologia , Mycobacterium tuberculosis/imunologia , Tuberculose Pulmonar/imunologia , Animais , Quimiocina CCL1/genética , Quimiocina CCL17/genética , Quimiocina CXCL12/genética , Células Dendríticas/microbiologia , Modelos Animais de Doenças , Regulação da Expressão Gênica , Interleucina-12/genética , Interleucina-23/genética , Pulmão/imunologia , Pulmão/patologia , Macrófagos/microbiologia , Camundongos , Camundongos Endogâmicos BALB C , Reação em Cadeia da Polimerase , Células Th1/imunologia , Células Th2/imunologia , Tuberculose Pulmonar/genéticaRESUMO
CD95 and CD95 ligand (CD95L) are critical molecules of the immune system. They play an important role in regulating T cell apoptosis. The membrane form of CD95L can be cleaved to release a soluble version, which possesses reduced apoptotic activity. In this study, we formally demonstrate that the reduced activity of soluble CD95L is due to its inability to oligomerize the CD95 receptor, an event that is critical for initiating death signaling from the receptor.
Assuntos
Apoptose , Peptídeos e Proteínas de Sinalização Intracelular , Glicoproteínas de Membrana/farmacologia , Linfócitos T/imunologia , Receptor fas/metabolismo , Proteína Reguladora de Apoptosis Semelhante a CASP8 e FADD , Proteínas de Transporte/metabolismo , Caspase 8 , Caspases/metabolismo , Células Clonais , Proteína Ligante Fas , Humanos , Glicoproteínas de Membrana/química , Transdução de Sinais , Linfócitos T/citologia , Linfócitos T/efeitos dos fármacosRESUMO
Receptors of the various cytokines although structurally diverse, can yet be grouped into four major families of receptor proteins. Most cytokines that function in the immune system bind to either the Class I or Class II receptor families. Two other important receptor families are the immunoglobulin superfamily receptor and the TNF receptor family. Members of these receptor families also have critical roles in the immune system. A common feature of all these receptor families is that they do not exhibit any intrinsic tyrosine kinase activity. Receptor signaling is initiated through recruitment of kinases and through recruitment of cytosolic proteins to the receptor. In this review we will examine receptor signaling pathways initiated from five receptors that are all involved in either initiating T helper-1 (Th1) responses, or in downregulating Th1 responses. The following receptors: Interleukin (IL)-12, Interferon (IFN), IL-4, IL-10, and Tumor necrosis factor (TNF)-alpha will be examined. Signaling initiated from IL-12, IFN-gamma and TNF-alpha are important for inducing Th1 responses, and on the other hand signaling from IL-4 and IL-10 receptors inhibit Th1 responses. We will also discuss human immunodeficiencies resulting from mutations in the genes that encode the Type I cytokine receptors.
Assuntos
Citocinas/metabolismo , Receptores de Citocinas/fisiologia , Transdução de Sinais/fisiologia , Células Th1/metabolismo , Animais , Humanos , Imunidade/fisiologia , Mutação , Infecções por Mycobacterium/imunologia , Receptores de Citocinas/genética , Receptores de Interferon/genética , Receptores de Interferon/fisiologia , Receptores de Interleucina/genética , Receptores de Interleucina/fisiologia , Receptores de Interleucina-12 , Receptor de Interferon gamaRESUMO
Thymic stromal lymphopoietin (TSLP) has been implicated in the development of allergic inflammation by promoting Th2-type responses and has become a potential therapeutic target. Using in vitro T cell differentiation cultures we were able to validate that TSLP played a more critical role in the early development of Th2 immune responses with less significant enhancement of already developed Th2 responses. Adoptive transfer of naive DO11.10 ovalbumin-specific T cells followed by airway exposure to ovalbumin showed an early impairment of Th2 immune response in TSLP-/- mice compared to wild type mice during the development of a Th2 response. In contrast, transfer of already differentiated Th2 cells into TSLP-/- mice did not change lung pathology or Th2 cytokine production upon ovalbumin challenge compared to transfer into wild type mice. An allergen-induced Th2 airway model demonstrated that there was only a difference in gob5 expression (a mucus-associated gene) between wild type and TSLP-/- mice. Furthermore, when allergic animals with established disease were treated with a neutralizing anti-TSLP antibody there was no change in airway hyperreponsiveness (AHR) or Th2 cytokine production compared to the control antibody treated animals, whereas a change in gob5 gene expression was also observed similar to the TSLP-/- mouse studies. In contrast, when animals were treated with anti-TSLP during the initial stages of allergen sensitization there was a significant change in Th2 cytokines during the final allergen challenge. Collectively, these studies suggest that in mice TSLP has an important role during the early development of Th2 immune responses, whereas its role at later stages of allergic disease may not be as critical for maintaining the Th2-driven allergic disease.
Assuntos
Alérgenos/imunologia , Diferenciação Celular/imunologia , Citocinas/metabolismo , Pneumopatias/imunologia , Pneumopatias/patologia , Células Th2/citologia , Células Th2/imunologia , Animais , Antígenos/imunologia , Baratas , Citocinas/biossíntese , Citocinas/deficiência , Células Dendríticas/imunologia , Hipersensibilidade/imunologia , Hipersensibilidade/patologia , Imunidade/imunologia , Linfonodos/imunologia , Linfonodos/patologia , Camundongos , Camundongos Endogâmicos BALB C , Transdução de Sinais/imunologia , Linfopoietina do Estroma do TimoRESUMO
In the present studies, we have established that RSV can elicit a more pathogenic environment dependent on improper DC-associated sensitization. Our initial studies demonstrated that RSV, but not influenza, infection during an allergen exposure into the airway induced a more severe allergen response. The RSV-induced exacerbation included an increased Th2 cytokine response and pathophysiology as monitored by AHR and mucus overproduction. DCs played a central role in the allergen-induced responses, as instilling RSV-infected BMDC into the airway could recapitulate a live virus challenge. With the use of CCR6-/- mice that have a primary defect in the recruitment of mDC subsets, reduced exacerbation of disease was observed when RSV was administered along with allergen. Furthermore, sensitization of mice with RSV-infected BMDC into the airway produced a more severe immune response to a live virus challenge. Subsequently, using RSV-infected BMDC from CCR7-/- mice (that do not migrate efficiently to LNs) to sensitize the exacerbated response demonstrated that the response was dependent on DC migration to the LN. Finally, the ability of RSV-infected DCs to elicit an exacerbated, allergen-induced pathogenic response could be maintained for as long as 3 weeks, suggesting that RSV-infected DCs themselves created an altered immune environment that impacts off-target mucosal responses that could have prolonged effects.
Assuntos
Alérgenos/imunologia , Movimento Celular/imunologia , Células Dendríticas/imunologia , Pneumopatias/etiologia , Receptores CCR6/fisiologia , Infecções por Vírus Respiratório Sincicial/imunologia , Vírus Sinciciais Respiratórios/imunologia , Animais , Feminino , Citometria de Fluxo , Pneumopatias/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos KnockoutRESUMO
The control of IL-12 production from dendritic cells (DCs) and macrophages in response to Mycobacterium tuberculosis (Mtb) is not well understood. The objective of this study was to pursue the mechanism underlying our previous report that in response to Mtb infection, DCs release abundant IL-12, whereas secretion is limited in macrophages. An initial comparison of IL-12p35 and IL-12p40 gene induction showed that p35 transcription is similar in murine bone marrow-derived DCs and macrophages, but a rapid and enhanced IL-12p40 transcription occurs only in DCs. Consistent with the p40 gene transcription profile, Mtb-induced remodeling at nucleosome 1 of the p40 promoter also occurs rapidly and extensively in DCs in comparison to macrophages. Removal of IL-10 or addition of IFNgamma enhances macrophage IL-12 release to Mtb, but without affecting the kinetics of remodeling at the macrophage p40 promoter. Furthermore, we show that Mtb-induced remodeling at the p40 promoter and IL-12 release in DCs is TLR9 dependent, and in contrast, TLR2 dependent, in macrophages. Data are also presented to demonstrate that a TLR9 agonist induces quantitatively more extensive remodeling at the IL-12p40 promoter and larger IL-12 release in comparison to a TLR2 agonist. Collectively, these findings suggest that DCs and macrophages handle Mtb differently resulting in only DCs being able to engage the more efficient TLR9 pathway for IL-12 gene induction. Our results also imply that TLR2 signaling is not a good inducer of IL-12, supporting the increasingly strong paradigm that TLR2 favors Th2 responses.
Assuntos
Células Dendríticas/imunologia , Interleucina-12/biossíntese , Macrófagos/imunologia , Receptor 2 Toll-Like/fisiologia , Receptor Toll-Like 9/fisiologia , Tuberculose/imunologia , Animais , MAP Quinases Reguladas por Sinal Extracelular/fisiologia , Subunidade p40 da Interleucina-12/genética , Camundongos , Camundongos Endogâmicos C57BL , Fosfatidilinositol 3-Quinases/fisiologia , Regiões Promotoras GenéticasRESUMO
Nucleotide-binding oligomerization domain proteins (NODs) are modular cytoplasmic proteins implicated in the recognition of peptidoglycan-derived molecules. NOD2 has recently been shown to be important for host cell cytokine responses to Mycobacterium tuberculosis, to synergize with Toll-like receptor 2 (TLR2) in mediating these responses, and thus to serve as a nonredundant recognition receptor for M. tuberculosis. Here, we demonstrate that macrophages and dendritic cells from NOD2-deficient mice were impaired in the production of proinflammatory cytokines and nitric oxide following infection with live, virulent M. tuberculosis. Mycolylarabinogalactan peptidoglycan (PGN), the cell wall core of M. tuberculosis, stimulated macrophages to release tumor necrosis factor (TNF) and interleukin-12p40 in a partially NOD2-dependent manner, and M. tuberculosis PGN required NOD2 for the optimal induction of TNF. However, NOD2-deficient mice were no more susceptible to infection with virulent M. tuberculosis than wild-type mice: they controlled the replication of M. tuberculosis in lung, spleen, and liver as well as wild-type mice, and both genotypes displayed similar lung pathologies. In addition, mice doubly deficient for NOD2 and TLR2 were similarly able to control an M. tuberculosis infection. Thus, NOD2 appears to participate in the recognition of M. tuberculosis by antigen-presenting cells in vitro yet is dispensable for the control of the pathogen during in vivo infection.
Assuntos
Mycobacterium tuberculosis/imunologia , Proteína Adaptadora de Sinalização NOD2/deficiência , Proteína Adaptadora de Sinalização NOD2/imunologia , Tuberculose/genética , Tuberculose/imunologia , Animais , Contagem de Colônia Microbiana , Citocinas/biossíntese , Células Dendríticas/imunologia , Subunidade p40 da Interleucina-12/biossíntese , Fígado/microbiologia , Pulmão/microbiologia , Pulmão/patologia , Macrófagos/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Mycobacterium tuberculosis/patogenicidade , Óxido Nítrico/biossíntese , Peptidoglicano/imunologia , Baço/microbiologia , Receptor 2 Toll-Like/deficiência , Fator de Necrose Tumoral alfa/genética , VirulênciaRESUMO
We previously reported that ligation of CD3 induces antiapoptotic signals in T helper 2 (Th2) cells, and in contrast causes Th1 cells to undergo apoptosis. Here we show that Cbl-b is accountable for the unequal response, revealing a previously unknown cell-specific regulatory function for the molecule. Absence of Cbl-b resulted in resistance to activation-induced apoptosis in murine Th1 cells following CD3 ligation, akin to what is observed in Th2 cells containing Cbl-b. Concurrent with the apoptosis profile, CD3 ligation in the absence of Cbl-b induced raft mobilization and cytoskeletal rearrangement in Th1 cells. Despite their ability to signal from CD3, Th2 cells did not aggregate their rafts, providing an explanation for cell-specific activity of Cbl-b.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/imunologia , Apoptose/imunologia , Proteínas Proto-Oncogênicas c-cbl/imunologia , Células Th1/imunologia , Células Th2/imunologia , Actinas/metabolismo , Animais , Complexo CD3/imunologia , Feminino , Microdomínios da Membrana/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Microscopia Confocal , Transdução de Sinais/imunologia , Células Th1/metabolismo , Células Th2/metabolismoRESUMO
The initial TLR-mediated interaction between Mycobacterium tuberculosis and dendritic cells is critical, since the cytokine production that ensues can greatly influence the class of adaptive immunity that is generated to the pathogen. In this study, we therefore determined the dependency on TLR2 and TLR4 for M. tuberculosis-induced cytokine production by murine dendritic cells. A key new finding of this study is that production of IL-6 and IL-10 from dendritic cells in response to M. tuberculosis is principally dependent on TLR2. The study also indicates that M. tuberculosis can induce IL-12 production in the absence of either TLR2 or TLR4, suggesting redundancy or possibly involvement of other receptors in IL-12 production. In addition, the data also reveal that lack of TLR2 or TLR4 does not impact on dendritic cell maturation or on their ability to influence the polarity of differentiating naive T cells. Collectively, data presented here provide a mechanistic insight for the contribution of TLR2 and TLR4 to tuberculosis disease progression and offer strategies for regulating IL-6 and IL-10 production in dendritic cell-based vaccine strategies.