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1.
J Med Microbiol ; 68(12): 1813-1822, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31674896

RESUMO

Introduction. Chronic pulmonary infection is associated with colonization with multiple micro-organisms but host-microbe and microbe-microbe interactions are poorly understood.Aim. This study aims to investigate the differences in host responses to mono- and co-infection with S. aureus and B. cenocepacia in human airway epithelial cells.Methodology. We assessed the effect of co-infection with B. cenocepacia and S. aureus on host signalling and inflammatory responses in the human airway epithelial cell line 16HBE, using ELISA and western blot analysis.Results. The results show that B. cenocepacia activates MAPK and NF-κB signalling pathways, subsequently eliciting robust interleukin (IL)-8 production. However, when airway epithelial cells were co-treated with live B. cenocepacia bacteria and S. aureus supernatants (conditioned medium), the pro-inflammatory response was attenuated. This anti-inflammatory effect was widely exhibited in the S. aureus isolates tested and was mediated via reduced MAPK and NF-κB signalling, but not via IL-1 receptor or tumour necrosis factor receptor modulation. The staphylococcal effectors were characterized as small, heat-stable, non-proteinaceous and not cell wall-related factors.Conclusion. This study demonstrates for the first time the host response in a S. aureus/B. cenocepacia co-infection model and provides insight into a staphylococcal immune evasion mechanism, as well as a therapeutic intervention for excessive inflammation.


Assuntos
Brônquios/imunologia , Infecções por Burkholderia/imunologia , Burkholderia cenocepacia/imunologia , Coinfecção/imunologia , Inflamação/etiologia , Infecções Estafilocócicas/imunologia , Células Cultivadas , Células Epiteliais/imunologia , Humanos , Evasão da Resposta Imune , Interleucina-8/biossíntese , Sistema de Sinalização das MAP Quinases/fisiologia , NF-kappa B/fisiologia
2.
Redox Biol ; 26: 101256, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31229842

RESUMO

Environmental proteases have been widely associated to the pathogenesis of allergic disorders. Der p 1, a cysteine-protease from house dust mite (HDM) Dermatophagoides pteronyssinus, constitutes one of the most clinically relevant indoor aeroallergens worldwide. Der p 1 protease activity depends on the redox status of its catalytic cysteine residue, which has to be in the reduced state to be active. So far, it is unknown whether Der p 1-protease activity could be regulated by host redox microenvironment once it reaches the lung epithelial lining fluid in addition to endogenous mite components. In this sense, Glutathione-S-transferase pi (GSTpi), an enzyme traditionally linked to phase II detoxification, is highly expressed in human lung epithelial cells, which represent the first line of defence against aeroallergens. Moreover, GSTpi is a generalist catalyst of protein S-glutathionylation reactions, and some polymorphic variants of this enzyme has been associated to the development of allergic asthma. Here, we showed that human GSTpi increased the cysteine-protease activity of Der p 1, while GSTmu (the isoenzyme produced by the mite) did not alter it. GSTpi induces the reduction of Cys residues in Der p 1, probably by rearranging its disulphide bridges. Furthermore, GSTpi was detected in the apical medium collected from human bronchial epithelial cell cultures, and more interesting, it increased cysteine-protease activity of Der p 1. Our findings support the role of human GSTpi from airways in modulating of Der p 1 cysteine-protease activity, which may have important clinical implications for immune response to this aeroallergen in genetically susceptible individuals.


Assuntos
Antígenos de Dermatophagoides/metabolismo , Proteínas de Artrópodes/metabolismo , Cisteína Endopeptidases/metabolismo , Cisteína/metabolismo , Dermatophagoides pteronyssinus/química , Células Epiteliais/enzimologia , Glutationa S-Transferase pi/metabolismo , Animais , Antígenos de Dermatophagoides/imunologia , Proteínas de Artrópodes/imunologia , Brônquios/citologia , Brônquios/enzimologia , Brônquios/imunologia , Linhagem Celular , Cisteína/imunologia , Cisteína Endopeptidases/imunologia , Dermatophagoides pteronyssinus/enzimologia , Dermatophagoides pteronyssinus/imunologia , Células Epiteliais/citologia , Células Epiteliais/imunologia , Glutationa S-Transferase pi/imunologia , Humanos , Isoenzimas/imunologia , Isoenzimas/metabolismo , Cinética , Oxirredução , Proteólise , Especificidade da Espécie
3.
Eur J Med Chem ; 175: 63-71, 2019 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-31075609

RESUMO

In the frame of a research program aimed to explore the relationship between chirality of iminosugars and their therapeutic potential, herein we report the synthesis of N-akyl l-deoxyiminosugars and the evaluation of the anti-inflammatory properties of selected candidates for the treatment of Pseudomonas aeruginosa infections in Cystic Fibrosis (CF) lung disease. Target glycomimetics were prepared by the shortest and most convenient approach reported to date, relying on the use of the well-known PS-TPP/I2 reagent system to prepare reactive alkoxyalkyl iodides, acting as key intermediates. Iminosugars ent-1-3 demonstrated to efficiently reduce the inflammatory response induced by P. aeruginosa in CuFi cells, either alone or in synergistic combination with their d-enantiomers, by selectively inhibiting NLGase. Surprisingly, the evaluation in murine models of lung disease showed that the amount of ent-1 required to reduce the recruitment of neutrophils was 40-fold lower than that of the corresponding d-enantiomer. The remarkably low dosage of the l-iminosugar, combined with its inability to act as inhibitor for most glycosidases, is expected to limit the onset of undesired effects, which are typically associated with the administration of its d-counterpart. Biological results herein obtained place ent-1 and congeners among the earliest examples of l-iminosugars acting as anti-inflammatory agents for therapeutic applications in Cystic Fibrosis.


Assuntos
Antibacterianos/uso terapêutico , Fibrose Cística/complicações , Imino Açúcares/uso terapêutico , Infecções por Pseudomonas/complicações , Infecções por Pseudomonas/tratamento farmacológico , Pseudomonas aeruginosa/isolamento & purificação , Animais , Antibacterianos/administração & dosagem , Antibacterianos/química , Antibacterianos/farmacologia , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/farmacologia , Brônquios/imunologia , Brônquios/microbiologia , Brônquios/patologia , Relação Dose-Resposta a Droga , Humanos , Imino Açúcares/administração & dosagem , Imino Açúcares/química , Imino Açúcares/farmacologia , Inflamação/prevenção & controle , Concentração Inibidora 50 , Camundongos , Neutrófilos/imunologia , Infecções por Pseudomonas/microbiologia , Pseudomonas aeruginosa/efeitos dos fármacos , Estereoisomerismo , beta-Glucosidase/antagonistas & inibidores
4.
Mol Immunol ; 112: 140-150, 2019 08.
Artigo em Inglês | MEDLINE | ID: mdl-31102986

RESUMO

The prevalence of fish allergy among fish-processing workers is higher than in the general population, possibly due to sensitization via inhalation and higher exposure. However, the response of the bronchial epithelium to fish allergens has never been explored. Parvalbumins (PVs) from bony fish are major sensitizers in fish allergy, while cartilaginous fish and their PVs are considered less allergenic. Increasing evidence demonstrates that components other than proteins from the allergen source, such as low molecular weight components smaller than 3 kDa (LMC) from pollen, may act as adjuvants during allergic sensitization. We investigated the response of bronchial epithelial cells to PVs and to LMC from Atlantic cod, a bony fish, and gummy shark, a cartilaginous fish. Polarized monolayers of the bronchial epithelial cell line 16HBE14o- were stimulated apically with fish PVs and/-or the corresponding fish LMC. Barrier integrity, transport of PVs across the monolayers and release of mediators were monitored. Intact PVs from both the bony and the cartilaginous fish were rapidly internalized by the cells and transported to the basolateral side of the monolayers. The PVs did not disrupt the epithelial barrier integrity nor did they modify the release of proinflammatory cytokines. In contrast, LMC from both fish species modified the physical and immunological properties of the epithelial barrier and the responses differed between bony and cartilaginous fish. While the barrier integrity was lowered by cod LMC 24 h after cell stimulation, it was increased by up to 2.3-fold by shark LMC. Furthermore, LMC from both fish species increased basolateral and apical release of IL-6 and IL-8, while CCL2 release was increased by cod but not by shark LMC. In summary, our study demonstrated the rapid transport of PVs across the epithelium which may result in their availability to antigen presenting cells required for allergic sensitization. Moreover, different cell responses to LMC derived from bony versus cartilaginous fish were observed, which may play a role in different allergenic potentials of these two fish classes.


Assuntos
Alérgenos/imunologia , Brônquios/imunologia , Citocinas/imunologia , Células Epiteliais/imunologia , Peixes/imunologia , Hipersensibilidade Alimentar/imunologia , Inflamação/imunologia , Animais , Linhagem Celular , Quimiocina CCL2/imunologia , Humanos , Interleucina-6/imunologia , Interleucina-8/imunologia , Peso Molecular , Parvalbuminas/imunologia , Alimentos Marinhos
5.
Nat Commun ; 10(1): 1856, 2019 04 23.
Artigo em Inglês | MEDLINE | ID: mdl-31015447

RESUMO

Bronchial premalignant lesions (PMLs) are precursors of lung squamous cell carcinoma, but have variable outcome, and we lack tools to identify and treat PMLs at risk for progression to cancer. Here we report the identification of four molecular subtypes of PMLs with distinct differences in epithelial and immune processes based on RNA-Seq profiling of endobronchial biopsies from high-risk smokers. The Proliferative subtype is enriched with bronchial dysplasia and exhibits up-regulation of metabolic and cell cycle pathways. A Proliferative subtype-associated gene signature identifies subjects with Proliferative PMLs from normal-appearing uninvolved large airway brushings with high specificity. In progressive/persistent Proliferative lesions expression of interferon signaling and antigen processing/presentation pathways decrease and immunofluorescence indicates a depletion of innate and adaptive immune cells compared with regressive lesions. Molecular biomarkers measured in PMLs or the uninvolved airway can enhance histopathological grading and suggest immunoprevention strategies for intercepting the progression of PMLs to lung cancer.


Assuntos
Biomarcadores Tumorais/genética , Carcinoma Broncogênico/patologia , Regulação Neoplásica da Expressão Gênica/imunologia , Neoplasias Pulmonares/patologia , Lesões Pré-Cancerosas/patologia , Antineoplásicos Imunológicos/farmacologia , Antineoplásicos Imunológicos/uso terapêutico , Biomarcadores Tumorais/imunologia , Biópsia , Brônquios/diagnóstico por imagem , Brônquios/imunologia , Brônquios/patologia , Broncoscopia , Carcinoma Broncogênico/genética , Carcinoma Broncogênico/imunologia , Carcinoma Broncogênico/prevenção & controle , Estudos de Coortes , Conjuntos de Dados como Assunto , Progressão da Doença , Detecção Precoce de Câncer/métodos , Perfilação da Expressão Gênica , Redes Reguladoras de Genes/genética , Redes Reguladoras de Genes/imunologia , Humanos , Imunidade Celular/efeitos dos fármacos , Imunidade Celular/genética , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/prevenção & controle , Programas de Rastreamento/métodos , Pessoa de Meia-Idade , Lesões Pré-Cancerosas/diagnóstico por imagem , Lesões Pré-Cancerosas/genética , Lesões Pré-Cancerosas/imunologia , RNA Mensageiro/genética , Mucosa Respiratória/citologia , Mucosa Respiratória/diagnóstico por imagem , Mucosa Respiratória/imunologia , Mucosa Respiratória/patologia , Análise de Sequência de RNA , Linfócitos T/imunologia , Tomografia Computadorizada por Raios X , Regulação para Cima
6.
J Immunol ; 202(9): 2519-2526, 2019 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-31010841

RESUMO

Inducible bronchus-associated lymphoid tissue (iBALT) is a tertiary lymphoid structure that resembles secondary lymphoid organs. iBALT is induced in the lung in response to Ag exposure. In some cases, such as infection with Mycobacterium tuberculosis, the formation of iBALT structure is indicative of an effective protective immune response. However, with persistent exposure to Ags during chronic inflammation, allergy, or autoimmune diseases, iBALT may be associated with exacerbation of inflammation. iBALT is characterized by well-organized T and B areas enmeshed with conventional dendritic cells, follicular dendritic cells, and stromal cells, usually located surrounding airways or blood vessels. Several of the molecular signals and cellular contributors that mediate formation of iBALT structures have been recently identified. This review will outline the recent findings associated with the formation and maintenance of iBALT and their contributions toward a protective or pathogenic function in pulmonary disease outcome.


Assuntos
Brônquios/imunologia , Células Dendríticas Foliculares/imunologia , Células Dendríticas/imunologia , Imunidade nas Mucosas , Pneumopatias/imunologia , Tecido Linfoide/imunologia , Animais , Brônquios/patologia , Células Dendríticas/patologia , Células Dendríticas Foliculares/patologia , Humanos , Pneumopatias/patologia , Tecido Linfoide/patologia
7.
Int Arch Allergy Immunol ; 179(1): 43-52, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30943513

RESUMO

BACKGROUND: The aim of this study was to investigate the role of Notch-1 signaling through Notch-1 ligands on bronchial epithelial cells (BECs) in regulating the development of T helper 2 (Th2) lymphocytes after RSV infection. METHODS: Firstly, we analyzed the expression of cytokines and Notch-1 ligands in BECs by using real-time PCR. Then, RSV-infected BECs were co-cultured with CD4+ T cells in a transwell chamber for 48 h, and differentiation of T cells in the lower chamber was determined using flow cytometry and real-time PCR. JAG1 siRNA was then used to determine the effects of Jagged/Notch-1 signaling on the differentiation of Th2. An RSV-infected mouse model was also used to analyze the secretion of Th differentiation-associated cytokines in serum and lung tissues using ELISA, the histopathological changes using HE staining, and the expression of JAG1 and JAG2 in BECs. RESULTS: The results showed that RSV promoted the expression of Th2-type cytokines and Jagged-1 and inhibited the expression of Jagged-2 in normal BECs. RSV-infected BECs induced Th2 differentiation. In addition, JAG1 downregulation inhibited the differentiation of Th2 and promoted differentiation of Th1. In the RSV-infected mouse model, the RSV titer, inflammation decreased with time. IL-4 and IL-17 increased on day 28 and 60, while IFNγ increased on day 7 and 28. Moreover, the expression of Jagged-1 increased and that of Jagged-2 decreased in BECs, which was consistent with IL-4 production in lung tissues. CONCLUSION: Our data showed that BECs had the potential to promote the differentiation of Th2 lymphocytes through Jagged-1/Notch-1 signaling.


Assuntos
Brônquios/fisiologia , Proteína Jagged-1/fisiologia , Proteína Jagged-2/fisiologia , Receptor Notch1/fisiologia , Infecções por Vírus Respiratório Sincicial/imunologia , Transdução de Sinais/fisiologia , Células Th2/citologia , Animais , Brônquios/imunologia , Brônquios/patologia , Diferenciação Celular , Citocinas/biossíntese , Células Epiteliais/fisiologia , Células HeLa , Humanos , Masculino , Camundongos , Camundongos Endogâmicos C57BL
8.
Basic Clin Pharmacol Toxicol ; 125(3): 304-314, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30861304

RESUMO

Reactive oxygen species (ROS) is regarded as a critical denominator in nanoparticle toxicology and inflammation. Previously, we have shown that silica nanoparticles sized 50 nm (Si50) induce release of CXCL8 and IL-6 from BEAS-2B cells, via mechanisms involving NFκB, p38 MAP kinase and TGF-α-activated EGF receptor. In the present study, the role of ROS-mediated mechanisms in the concentration-dependent Si50 induction of CXCL8 and IL-6 responses was examined. Si50 (200 µg/mL) induced a time-dependent ROS formation and a postponed increase in expression of haem oxygenase (HO-1) mRNA and protein. Pre-treatment with the ROS inhibitors N-acetyl cysteine (NAC) and diphenyleneiodonium (DPI) partially attenuated CXCL8 and IL-6 responses to 200 µg/mL, but not to 100 µg/mL Si50. The release of TGF-α induced by Si50 (200 µg/mL) was significantly reduced by NAC, but not by DPI nor siRNA against NADPH oxidase DUOX-1 (siDUOX-1). Furthermore, siDUOX-1 reduced Si50-induced CXCL8, but not IL-6. Both p38 and p65 phosphorylations were inhibited by siDUOX-1, but for NAC only p65 phosphorylation reached a significant reduction. Neither NAC nor DPI reduced Si50-induced CXCL8 and IL-6 gene expressions. In conclusion, Si50-induced CXCL8 and IL-6 involved both ROS-dependent and ROS-independent mechanisms. Notably, the role of ROS seemed restricted to effects of higher concentrations of Si50 and not mediated via the gene expression.


Assuntos
Brônquios/efeitos dos fármacos , Nanopartículas/toxicidade , Espécies Reativas de Oxigênio/metabolismo , Mucosa Respiratória/efeitos dos fármacos , Dióxido de Silício/toxicidade , Brônquios/citologia , Brônquios/imunologia , Linhagem Celular , Oxidases Duais/genética , Oxidases Duais/metabolismo , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/imunologia , Regulação da Expressão Gênica/efeitos dos fármacos , Regulação da Expressão Gênica/imunologia , Humanos , Interleucina-6/imunologia , Interleucina-6/metabolismo , Interleucina-8/imunologia , Interleucina-8/metabolismo , Proteína Quinase 14 Ativada por Mitógeno/metabolismo , Tamanho da Partícula , Fosforilação/efeitos dos fármacos , Fosforilação/genética , RNA Interferente Pequeno/metabolismo , Espécies Reativas de Oxigênio/antagonistas & inibidores , Espécies Reativas de Oxigênio/imunologia , Mucosa Respiratória/citologia , Mucosa Respiratória/imunologia , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologia , Fator de Transcrição RelA/metabolismo
9.
Immunity ; 50(3): 707-722.e6, 2019 03 19.
Artigo em Inglês | MEDLINE | ID: mdl-30824323

RESUMO

Type 2 lymphocytes promote both physiologic tissue remodeling and allergic pathology, yet their physical tissue niches are poorly described. Here, we used quantitative imaging to define the tissue niches of group 2 innate lymphoid cells (ILC2s), which are critical instigators of type 2 immunity. We identified a dominant adventitial niche around lung bronchi and larger vessels in multiple tissues, where ILC2s localized with subsets of dendritic and regulatory T cells. However, ILC2s were most intimately associated with adventitial stromal cells (ASCs), a mesenchymal fibroblast-like subset that expresses interleukin-33 (IL-33) and thymic stromal lymphopoietin (TSLP). In vitro, ASCs produced TSLP that supported ILC2 accumulation and activation. ILC2s and IL-13 drove reciprocal ASC expansion and IL-33 expression. During helminth infection, ASC depletion impaired lung ILC2 and Th2 cell accumulation and function, which are in part dependent on ASC-derived IL-33. These data indicate that adventitial niches are conserved sites where ASCs regulate type 2 lymphocyte expansion and function.


Assuntos
Imunidade Inata/imunologia , Linfócitos/imunologia , Células Estromais/imunologia , Animais , Brônquios/imunologia , Citocinas/imunologia , Interleucina-13/imunologia , Interleucina-33/imunologia , Camundongos , Linfócitos T Reguladores/imunologia , Células Th2/imunologia
10.
Artigo em Inglês | MEDLINE | ID: mdl-30744811

RESUMO

We investigated the mechanism responsible for the oxidative stress and pro-inflammatory response triggered by PM2.5 collected from Nanjing of China. Two human bronchial epithelia cell lines (HBE and BEAS-2B) were used. Human gene expression profile microarray was performed to investigate the alteration of gene expression in PM2.5-treated HBE cells. The results of ROS assay and ELISA indicated that PM2.5 (150 µg/ml) increased the level of cellular reactive oxygen species (ROS) and promoted the release of interleukin-6 (IL-6) in HBE cells. CYP1A1 and CYP1B1 were the top two up-regulated genes by PM2.5 (150 µg/ml, 48 h of exposure) in HBE cells. Co-knockdown of CYP1A1/1B1 by siRNA substantially inhibited PM2.5-induced ROS generation, IL-6/IL-8 secretion and STAT3/P-STAT3 expression. Similarly, the knockdown of STAT3 also effectively inhibited PM2.5-induced rise in ROS level and IL-6/IL-8 secretion. In summary, PM2.5 mediated oxidative stress and pro-inflammatory response via up-regulating the expression of CYP1A1/1B1 in two human bronchial epithelial cell lines.


Assuntos
Brônquios/patologia , Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP1B1/metabolismo , Células Epiteliais/patologia , Mediadores da Inflamação/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Material Particulado/farmacologia , Poluentes Atmosféricos/farmacologia , Brônquios/efeitos dos fármacos , Brônquios/imunologia , Brônquios/metabolismo , Células Cultivadas , Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1B1/genética , Células Epiteliais/efeitos dos fármacos , Células Epiteliais/imunologia , Células Epiteliais/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Técnicas In Vitro , Espécies Reativas de Oxigênio
11.
Part Fibre Toxicol ; 16(1): 8, 2019 02 13.
Artigo em Inglês | MEDLINE | ID: mdl-30760282

RESUMO

BACKGROUND: It is well established that toxicological evaluation of engineered nanomaterials (NMs) is vital to ensure the health and safety of those exposed to them. Further, there is a distinct need for the development of advanced physiologically relevant in vitro techniques for NM hazard prediction due to the limited predictive power of current in vitro models and the unsustainability of conducting nano-safety evaluations in vivo. Thus, the purpose of this study was to develop alternative in vitro approaches to assess the potential of NMs to induce genotoxicity by secondary mechanisms. RESULTS: This was first undertaken by a conditioned media-based technique, whereby cell culture media was transferred from differentiated THP-1 (dTHP-1) macrophages treated with γ-Fe2O3 or Fe3O4 superparamagnetic iron oxide nanoparticles (SPIONs) to the bronchial cell line 16HBE14o-. Secondly construction and SPION treatment of a co-culture model comprising of 16HBE14o- cells and dTHP-1 macrophages. For both of these approaches no cytotoxicity was detected and chromosomal damage was evaluated by the in vitro micronucleus assay. Genotoxicity assessment was also performed using 16HBE14o- monocultures, which demonstrated only γ-Fe2O3 nanoparticles to be capable of inducing chromosomal damage. In contrast, immune cell conditioned media and dual cell co-culture SPION treatments showed both SPION types to be genotoxic to 16HBE14o- cells due to secondary genotoxicity promoted by SPION-immune cell interaction. CONCLUSIONS: The findings of the present study demonstrate that the approach of using single in vitro cell test systems precludes the ability to consider secondary genotoxic mechanisms. Consequently, the use of multi-cell type models is preferable as they better mimic the in vivo environment and thus offer the potential to enhance understanding and detection of a wider breadth of potential damage induced by NMs.


Assuntos
Dano ao DNA , Compostos Férricos/toxicidade , Nanopartículas de Magnetita/toxicidade , Testes de Mutagenicidade/métodos , Brônquios/efeitos dos fármacos , Brônquios/imunologia , Brônquios/patologia , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/genética , Sobrevivência Celular/efeitos dos fármacos , Técnicas de Cocultura , Meios de Cultivo Condicionados , Citocinas/biossíntese , Endocitose/efeitos dos fármacos , Humanos , Técnicas In Vitro , Macrófagos/efeitos dos fármacos , Macrófagos/imunologia , Macrófagos/patologia , Células THP-1
12.
Viruses ; 11(1)2019 01 11.
Artigo em Inglês | MEDLINE | ID: mdl-30641955

RESUMO

Immunopathologic examination of the lungs of mouse models of experimental influenza virus infection provides new insights into the immune response in this disease. First, there is rapidly developing perivascular and peribronchial infiltration of the lung with T-cells. This is followed by invasion of T-cells into the bronchiolar epithelium, and separation of epithelial cells from each other and from the basement membrane leading to defoliation of the bronchial epithelium. The intraepithelial reaction may involve either CD8 or CD4 T-cytotoxic cells and is analogous to a viral exanthema of the skin, such as measles and smallpox, which occur when the immune response against these infections is activated and the infected cells are attacked by T-cytotoxic cells. Then there is formation of B-cell follicles adjacent to bronchi, i.e., induced bronchial associated lymphoid tissue (iBALT). iBALT reacts like the cortex of a lymph node and is a site for a local immune response not only to the original viral infection, but also related viral infections (heterologous immunity). Proliferation of Type II pneumocytes and/or terminal bronchial epithelial cells may extend into the adjacent lung leading to large zones filled with tumor-like epithelial cells. The effective killing of influenza virus infected epithelial cells by T-cytotoxic cells and induction of iBALT suggests that adding the induction of these components might greatly increase the efficacy of influenza vaccination.


Assuntos
Modelos Animais de Doenças , Vacinas contra Influenza/imunologia , Infecções por Orthomyxoviridae/imunologia , Linfócitos T Citotóxicos/imunologia , Linfócitos T Reguladores/imunologia , Animais , Anticorpos Antivirais/sangue , Brônquios/citologia , Brônquios/imunologia , Brônquios/virologia , Proliferação de Células , Desenho de Drogas , Imunidade nas Mucosas , Camundongos , Orthomyxoviridae
13.
Basic Clin Pharmacol Toxicol ; 125(1): 8-15, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-30697954

RESUMO

There has been considerable research on the involvement of RhoA/Rho kinase signalling in smooth muscle contractions. However, only a few reports have addressed the specific role of Rac1, which is a member of the Rho GTPase superfamily. Therefore, this study investigated the role of Rac1-related pathways in bronchial smooth muscle (BSM) contractions. Bronchial rings isolated from mice were suspended in an organ bath, and the isometric contractions of circular smooth muscles were monitored. The phosphorylation of myosin light chains (MLCs) was analysed by immunoblotting. The Rac1 inhibitor EHT1864 inhibited carbachol (CCh)-induced BSM contractions, although high K+ depolarization-induced BSM contractions were not significantly attenuated by EHT1864. Moreover, high K+ - and phorbol 12,13-dibutyrate (PDBu; PKC activator)-induced contractions were not attenuated by Rac1 inhibition, whereas sodium fluoride (NaF)-induced force development was inhibited by EHT1864. The gene and protein expression of Rac1 was increased in the BSM of a murine model with antigen-induced airway hyper-responsiveness (AHR). In addition, an increased force of the BSM contractions in AHR was suppressed by EHT1864 treatment, suggesting that the up-regulation of Rac1 is involved in AHR. These findings suggest that an increase in Rac1-mediated signalling is involved in the augmented contractions of BSMs in antigen-induced AHR mice.


Assuntos
Asma/patologia , Brônquios/patologia , Contração Muscular/imunologia , Músculo Liso/patologia , Neuropeptídeos/metabolismo , Proteínas rac1 de Ligação ao GTP/metabolismo , Animais , Asma/imunologia , Brônquios/efeitos dos fármacos , Brônquios/imunologia , Carbacol/farmacologia , Modelos Animais de Doenças , Humanos , Masculino , Camundongos , Contração Muscular/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Músculo Liso/imunologia , Neuropeptídeos/antagonistas & inibidores , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Pironas/farmacologia , Quinolinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Regulação para Cima , Proteínas rac1 de Ligação ao GTP/antagonistas & inibidores
14.
Am J Physiol Lung Cell Mol Physiol ; 316(1): L71-L81, 2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-30335498

RESUMO

Corticosteroids (CSs) are commonly used to manage wheezing and asthma in pediatric populations. Although corticosteroids are effective in alleviating airway diseases, some children with more moderate-severe asthma phenotypes show CS resistance and exhibit significant airflow obstruction, persistent inflammation, and more frequent exacerbations. Previous studies have demonstrated that Th1 cytokines, such as TNF-α and IFN-γ, promote CS resistance in adult human airway smooth muscle (ASM). In the present study, using a human fetal ASM cell model, we tested the hypothesis that TNF-α/IFN-γ induces CS resistance. In contrast to TNF-α or IFN-γ alone, the combination of TNF-α/IFN-γ blunted the ability of fluticasone propionate (FP) to reduce expression of the chemokines CCL5 and CXCL10 despite expression of key anti-inflammatory glucocorticoid receptor target genes being largely unaffected by TNF-α/IFN-γ. Expression of the NF-κB subunit p65 and phosphorylation of Stat1 were elevated in cells treated with TNF-α/IFN-γ, an effect that remained in the presence of FP. siRNA knockdown studies demonstrated the effects of TNF-α/IFN-γ on increased p65 are mediated by Stat1, a transcription factor activated by IFN-γ. Expression of TNFAIP3, a negative regulator of NF-κB activity, was not altered by TNF-α/IFN-γ. However, the effects of TNF-α/IFN-γ were partially reduced by overexpression of TNFAIP3 but did not influence p65 expression. Together, these data suggest that IFN-γ augments the effects of TNF-α on chemokines by enhancing expression of key inflammatory pathways in the presence of CS. Interactions between TNF-α- and IFN-γ-mediated pathways may promote inflammation in asthmatic children resistant to CSs.


Assuntos
Corticosteroides/farmacologia , Brônquios/imunologia , Resistência a Medicamentos/efeitos dos fármacos , Fluticasona/farmacologia , Interferon gama/imunologia , Miócitos de Músculo Liso/imunologia , Traqueia/imunologia , Fator de Necrose Tumoral alfa/imunologia , Brônquios/crescimento & desenvolvimento , Criança , Pré-Escolar , Resistência a Medicamentos/imunologia , Feminino , Humanos , Masculino , Células Th1/imunologia , Traqueia/crescimento & desenvolvimento
16.
Mol Immunol ; 105: 205-212, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30553057

RESUMO

Carbon monoxide (CO) is an anti-inflammatory gaseous molecule produced endogenously by heme oxygenases (HOs) HO-1 and HO-2. However, the mechanisms underlying the anti-inflammatory effects of CO in the human bronchial epithelium are still not fully understood. In this study, the cationic peptide poly-l-arginine (PLA) was utilized to induce bronchial epithelial damage and subsequent pro-inflammatory cytokine release in the human bronchial epithelial cell line 16HBE14o-. Expression of both HO-1 and HO-2 after PLA exposure was examined. The polarized secretion of two pro-inflammatory cytokines, interleukin (IL)-6 and IL-8, was determined by ELISA. The anti-inflammatory effects of CO liberated from CO-releasing molecules (CORMs) were examined by both ELISA and western blot analysis. Our results indicate that PLA exposure leads to upregulation of HO-1 expression and p65 NF-κB phosphorylation, as well as IL-6 and IL-8 release. HO-1 induction by hemin or CORMs significantly suppressed IL-6 and IL-8 release. In addition, HO-1 knockdown further increased IL-6 and IL-8 release under basal and PLA-stimulated conditions. Our results thereby demonstrate that the HO-1/CO axis exerts significant anti-inflammatory activity during bronchial epithelial damage caused by cationic protein.


Assuntos
Anti-Inflamatórios/farmacologia , Brônquios/imunologia , Monóxido de Carbono/farmacologia , Heme Oxigenase-1/imunologia , Peptídeos/farmacologia , Mucosa Respiratória/imunologia , Linhagem Celular , Heme Oxigenase (Desciclizante)/genética , Heme Oxigenase (Desciclizante)/imunologia , Heme Oxigenase-1/genética , Humanos , Interleucina-6/genética , Interleucina-6/imunologia , Interleucina-8/genética , Interleucina-8/imunologia
17.
J Allergy Clin Immunol ; 143(1): 114-125.e4, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-29698627

RESUMO

BACKGROUND: The innate immune system senses viral infection through pattern recognition receptors (PRRs), leading to type I interferon production. The role of type I interferon and PPRs in rhinovirus-induced asthma exacerbations in vivo are uncertain. OBJECTIVES: We sought to compare bronchial mucosal type I interferon and PRR expression at baseline and after rhinovirus infection in atopic asthmatic patients and control subjects. METHODS: Immunohistochemistry was used to detect expression of IFN-α, IFN-ß, and the PRRs: Toll-like receptor 3, melanoma differentiation-associated gene 5, and retinoic acid-inducible protein I in bronchial biopsy specimens from 10 atopic asthmatic patients and 15 nonasthmatic nonatopic control subjects at baseline and on day 4 and 6 weeks after rhinovirus infection. RESULTS: We observed IFN-α/ß deficiency in the bronchial epithelium at 3 time points in asthmatic patients in vivo. Lower epithelial IFN-α/ß expression was related to greater viral load, worse airway symptoms, airway hyperresponsiveness, and reductions in lung function during rhinovirus infection. We found lower frequencies of bronchial subepithelial monocytes/macrophages expressing IFN-α/ß in asthmatic patients during infection. Interferon deficiency at baseline was not accompanied by deficient PRR expression in asthmatic patients. Both epithelial and subepithelial PRR expression were induced during rhinovirus infection. Rhinovirus infection-increased numbers of subepithelial interferon/PRR-expressing inflammatory cells were related to greater viral load, airway hyperresponsiveness, and reductions in lung function. CONCLUSIONS: Bronchial epithelial IFN-α/ß expression and numbers of subepithelial IFN-α/ß-expressing monocytes/macrophages during infection were both deficient in asthmatic patients. Lower epithelial IFN-α/ß expression was associated with adverse clinical outcomes after rhinovirus infection in vivo. Increases in numbers of subepithelial cells expressing interferon/PRRs during infection were also related to greater viral load/illness severity.


Assuntos
Asma/imunologia , Proteína DEAD-box 58/imunologia , Regulação da Expressão Gênica/imunologia , Helicase IFIH1 Induzida por Interferon/biossíntese , Interferon-alfa/imunologia , Interferon beta/imunologia , Infecções por Picornaviridae/imunologia , Rhinovirus/imunologia , Receptor 3 Toll-Like/imunologia , Adulto , Asma/metabolismo , Asma/patologia , Biópsia , Brônquios/imunologia , Brônquios/metabolismo , Brônquios/patologia , Proteína DEAD-box 58/biossíntese , Feminino , Humanos , Helicase IFIH1 Induzida por Interferon/imunologia , Interferon-alfa/metabolismo , Interferon beta/metabolismo , Masculino , Infecções por Picornaviridae/metabolismo , Infecções por Picornaviridae/patologia , Rhinovirus/metabolismo , Índice de Gravidade de Doença , Receptor 3 Toll-Like/biossíntese
18.
J Virol ; 93(2)2019 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-30333178

RESUMO

Rhinoviral infection is a common trigger of the excessive inflammation observed during exacerbations of asthma and chronic obstructive pulmonary disease. Rhinovirus (RV) recognition by pattern recognition receptors activates the mitogen-activated protein kinase (MAPK) pathways, which are common inducers of inflammatory gene production. A family of dual-specificity phosphatases (DUSPs) can regulate MAPK function, but their roles in rhinoviral infection are not known. We hypothesized that DUSPs would negatively regulate the inflammatory response to RV infection. Our results revealed that the p38 and c-Jun N-terminal kinase (JNK) MAPKs play key roles in the inflammatory response of epithelial cells to RV infection. Three DUSPs previously shown to have roles in innate immunity (DUSPs 1, 4, and 10) were expressed in primary bronchial epithelial cells, and one of them, DUSP10, was downregulated by RV infection. Small interfering RNA-mediated knockdown of DUSP10 identified a role for the protein in negatively regulating inflammatory cytokine production in response to interleukin-1ß (IL-1ß), alone and in combination with RV, without any effect on RV replication. This study identifies DUSP10 as an important regulator of airway inflammation in respiratory viral infection.IMPORTANCE Rhinoviruses are one of the causes of the common cold. In patients with asthma or chronic obstructive pulmonary disease, viral infections, including those with rhinovirus, are the commonest cause of exacerbations. Novel therapeutics to limit viral inflammation are clearly required. The work presented here identifies DUSP10 as an important protein involved in limiting the inflammatory response in the airway without affecting immune control of the virus.


Assuntos
Brônquios/virologia , Fosfatases de Especificidade Dupla/metabolismo , Interleucina-1beta/farmacologia , Fosfatases da Proteína Quinase Ativada por Mitógeno/metabolismo , Rhinovirus/patogenicidade , Brônquios/citologia , Brônquios/imunologia , Células Cultivadas , Regulação para Baixo , Fosfatases de Especificidade Dupla/genética , Células Epiteliais/citologia , Células Epiteliais/imunologia , Células Epiteliais/virologia , Técnicas de Silenciamento de Genes , Humanos , Sistema de Sinalização das MAP Quinases , Fosfatases da Proteína Quinase Ativada por Mitógeno/genética , Rhinovirus/imunologia
19.
J Clin Invest ; 129(2): 556-568, 2019 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-30561386

RESUMO

Antibody-mediated rejection (AMR) is a principal cause of acute and chronic failure of lung allografts. However, mechanisms mediating this oftentimes fatal complication are poorly understood. Here, we show that Foxp3+ T cells formed aggregates in rejection-free human lung grafts and accumulated within induced bronchus-associated lymphoid tissue (BALT) of tolerant mouse lungs. Using a retransplantation model, we show that selective depletion of graft-resident Foxp3+ T lymphocytes resulted in the generation of donor-specific antibodies (DSA) and AMR, which was associated with complement deposition and destruction of airway epithelium. AMR was dependent on graft infiltration by B and T cells. Depletion of graft-resident Foxp3+ T lymphocytes resulted in prolonged interactions between B and CD4+ T cells within transplanted lungs, which was dependent on CXCR5-CXCL13. Blockade of CXCL13 as well as inhibition of the CD40 ligand and the ICOS ligand suppressed DSA production and prevented AMR. Thus, we have shown that regulatory Foxp3+ T cells residing within BALT of tolerant pulmonary allografts function to suppress B cell activation, a finding that challenges the prevailing view that regulation of humoral responses occurs peripherally. As pulmonary AMR is largely refractory to current immunosuppression, our findings provide a platform for developing therapies that target local immune responses.


Assuntos
Citotoxicidade Celular Dependente de Anticorpos , Linfócitos B , Brônquios , Rejeição de Enxerto , Transplante de Pulmão , Ativação Linfocitária , Animais , Linfócitos B/imunologia , Linfócitos B/patologia , Brônquios/imunologia , Brônquios/patologia , Ligante de CD40/genética , Ligante de CD40/imunologia , Quimiocina CXCL13/genética , Quimiocina CXCL13/imunologia , Rejeição de Enxerto/genética , Rejeição de Enxerto/imunologia , Rejeição de Enxerto/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Knockout , Camundongos Nus , Receptores CXCR5/genética , Receptores CXCR5/imunologia , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/patologia
20.
PLoS One ; 13(11): e0208277, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30500862

RESUMO

Mouse models have been essential to generate supporting data for the research of infectious diseases. Burkholderia pseudomallei, the etiological agent of melioidosis, has been studied using mouse models to investigate pathogenesis and efficacy of novel medical countermeasures to include both vaccines and therapeutics. Previous characterization of mouse models of melioidosis have demonstrated that BALB/c mice present with an acute infection, whereas C57BL/6 mice have shown a tendency to be more resistant to infection and may model chronic disease. In this study, either BALB/c or C57BL/6 mice were exposed to aerosolized human clinical isolates of B. pseudomallei. The bacterial strains included HBPUB10134a (virulent isolate from Thailand), MSHR5855 (virulent isolate from Australia), and 1106a (relatively attenuated isolate from Thailand). The LD50 values were calculated and serial sample collections were performed in order to examine the bacterial burdens in tissues, histopathological features of disease, and the immune response mounted by the mice after exposure to aerosolized B. pseudomallei. These data will be important when utilizing these models for testing novel medical countermeasures. Additionally, by comparing highly virulent strains with attenuated isolates, we hope to better understand the complex disease pathogenesis associated with this bacterium.


Assuntos
Burkholderia pseudomallei/fisiologia , Melioidose/patologia , Animais , Formação de Anticorpos , Austrália/epidemiologia , Brônquios/imunologia , Brônquios/microbiologia , Brônquios/patologia , Burkholderia pseudomallei/patogenicidade , Citocinas/sangue , Citocinas/imunologia , Modelos Animais de Doenças , Progressão da Doença , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Melioidose/sangue , Melioidose/epidemiologia , Melioidose/imunologia , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Tailândia/epidemiologia , Virulência
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