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Thorsten Dürk,
Daniel Duerschmied,
Tobias Müller,
Melanie Grimm,
Sebastian Reuter,
Rodolfo Paula Vieira,
Korcan Ayata,
Sanja Cicko, Stephan Sorichter,
Diego J Walther,
J Christian Virchow,
Christian Taube,
Marco Idzko
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ABSTRACT: RATIONALE: 5-hydroyxtryptamine (5-HT) is involved in the pathogenesis of allergic airway inflammation (AAI). It is unclear, however, how 5-HT contributes to AAI and whether this depends on tryptophan hydroxylase (TPH) 1, the critical enzyme for peripheral 5-HT synthesis. OBJECTIVES: To elucidate the role TPH1 and the peripheral source of 5-HT in asthma pathogenesis. METHODS: TPH1-deficient and TPH1-inhibitor-treated animals were challenged in OVA-alum and HDM-models of allergic airway inflammation (AAI). Experiments with bone-marrow chimera, mast cell-deficient animals, platelets transfusion, and bone-marrow dendritic cells (BMDC)-driven model of AAI were performed. 5-HT-levels were measured in bronchoalveolar fluid (BALF) and/or serum of animals with AAI and in human asthma. MEASUREMENTS AND MAIN RESULTS: 5-HT-levels are increased in BALF of mice and human asthmatics following allergen provocation. TPH1-deficiency and TPH1-inhibition reduced all cardinal features of AAI. Administration of exogenous 5-HT restored AAI in TPH1-deficient mice. The pivotal role of 5-HT-production by structural cells was corroborated by BM-chimera experiments. Experiments in mast cell-deficient mice revealed that mast cells are not a source of 5-HT while transfusion of platelets from WT and TPH1-deficient mice revealed that only platelets containing 5-HT enhanced AAI. Lack of endogenous 5-HT in vitro and in vivo was associated with an impaired Th2-priming capacity of BMDC. CONCLUSION: In summary, TPH1-deficiency or -inhibition reduces AAI. Platelet- and not mast cell-derived 5-HT is pivotal in AAI, and lack of 5-HT leads to an impaired Th2-priming capacity of BMDC. Thus, targeting TPH1 could offer novel therapeutic options for asthma.
American Journal of Respiratory and Critical Care Medicine 01/2013; · 11.08 Impact Factor
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Elisabeth Panther,
Thorsten Dürk,
Davide Ferrari,
Francesco Di Virgilio,
Melanie Grimm, Stephan Sorichter,
Sanja Cicko,
Yared Herouy,
Johannes Norgauer,
Marco Idzko,
Tobias Müller
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ABSTRACT: The nucleotide adenosine-5'-monophosphate (AMP) can be released by various cell types and has been shown to elicit different cellular responses. In the extracellular space AMP is dephosphorylated to the nucleoside adenosine which can then bind to adenosine receptors. However, it has been shown that AMP can also activate A(1) and A(2a) receptors directly. Here we show that AMP is a potent modulator of mouse and human dendritic cell (DC) function. AMP increased intracellular Ca(2+) concentration in a time and dose dependent manner. Furthermore, AMP stimulated actin-polymerization in human DCs and induced migration of immature human and bone marrow derived mouse DCs, both via direct activation of A(1) receptors. AMP strongly inhibited secretion of TNF-α and IL-12p70, while it enhanced production of IL-10 both via activation of A(2a) receptors. Consequently, DCs matured in the presence of AMP and co-cultivated with naive CD4(+)CD45RA(+) T cells inhibited IFN-γ production whereas secretion of IL-5 and IL-13 was up-regulated. An enhancement of Th2-driven immune response could also be observed when OVA-pulsed murine DCs were pretreated with AMP prior to co-culture with OVA-transgenic naïve OTII T cells. An effect due to the enzymatic degradation of AMP to adenosine could be ruled out, as AMP still elicited migration and changes in cytokine secretion in bone-marrow derived DCs generated from CD73-deficient animals and in human DCs pretreated with the ecto-nucleotidase inhibitor 5'-(alpha,beta-methylene) diphosphate (APCP). Finally, the influence of contaminating adenosine could be excluded, as AMP admixed with adenosine desaminase (ADA) was still able to influence DC function. In summary our data show that AMP when present during maturation is a potent regulator of dendritic cell function and point out the role for AMP in the pathogenesis of inflammatory disorders.
PLoS ONE 01/2012; 7(5):e37560. · 4.09 Impact Factor
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Rodolfo Paula Vieira,
Tobias Müller,
Melanie Grimm,
Volker von Gernler,
Britta Vetter,
Thorsten Dürk,
Sanja Cicko,
C Korcan Ayata, Stephan Sorichter,
Bernard Robaye,
Robert Zeiser,
Davide Ferrari,
Andreas Kirschbaum,
Gernot Zissel,
J Christian Virchow,
Jean-Marie Boeynaems,
Marco Idzko
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ABSTRACT: Extracellular nucleotides have recently been identified as proinflammatory mediators involved in asthma pathogenesis by signaling via purinergic receptors, but the role of the purinergic receptor type 6 (P2Y6R) has not been previously investigated.
To investigate the role of P2Y6R in asthma pathogenesis.
Acute and chronic OVA model and also HDM model of allergic inflammation in C57Bl/6 mice treated with specific P2Y6R antagonist and P2Y6R(-/-) mice were evaluated for classical features of asthmatic inflammation. In addition, primary epithelial cell culture from human and epithelial cell lines from mouse and human were stimulated with P2Y6R agonist and treated with P2Y6R antagonist and assessed for IL-6, IL-8/CXCL8 and KC levels. Experiments with P2Y6R(-/-) and P2Y6R(+/+) chimera were performed to discriminate the role of P2Y6R activation in structural lung cells and in cells from hematopoietic system.
We observed that the intratracheal application of a P2Y6R antagonist (MRS2578) and P2Y6R deficiency inhibited cardinal features of asthma, such as bronchoalveolar lavage eosinophilia, airway remodeling, Th2 cytokine production, and bronchial hyperresponsiveness in the ovalbumin-alum model. MRS2578 was also effective in reducing airway inflammation in a model using house dust mite extracts to induce allergic lung inflammation. Experiments with bone marrow chimeras revealed the importance of the P2Y6R expression on lung structural cells in airway inflammation. In accordance with this finding, we found a strong up-regulation of P2Y6 expression on airway epithelial cells of animals with experimental asthma. Concerning the underlying mechanism, we observed that MRS2578 inhibited the release of IL-6 and IL-8/KC by lung epithelial cells in vivo, whereas intrapulmonary application of the P2Y6R agonist uridine-5'-diphosphate increased the bronchoalveolar levels of IL-6 and KC. In addition, selective activation of P2Y6 receptors induced the release of IL-6 and KC/IL-8 by murine and human lung epithelial cells in vitro.
P2Y6R expression on airway epithelial cells is up-regulated during acute and chronic allergic airway inflammation, and selective blocking of P2Y6R or P2Y6R deficiency on the structural cells reduces cardinal features of experimental asthma. Thus, blocking pulmonary P2Y6R might be a target for the treatment of allergic airway inflammation.
American Journal of Respiratory and Critical Care Medicine 04/2011; 184(2):215-23. · 11.08 Impact Factor
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Tobias Müller,
Rodolfo Paula Vieira,
Melanie Grimm,
Thorsten Dürk,
Sanja Cicko,
Robert Zeiser,
Thilo Jakob,
Stefan F Martin,
Britta Blumenthal, Stephan Sorichter,
Davide Ferrari,
Francesco Di Virgillio,
Marco Idzko
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ABSTRACT: P2X₇R deficiency is associated with a less severe outcome in acute and chronic inflammatory disorders. Recently, we demonstrated that extracellular adenosine triphosphate is involved in the pathogenesis of asthma by modulating the function of dendritic cells (DCs). However, the role of the purinergic receptor subtype P2X₇ is unknown. To elucidate the role of P2X₇R in allergic airway inflammation (AAI) in vitro and in vivo, P2X₇R expression was measured in lung tissue and immune cells of mice or in humans with allergic asthma. By using a specific P2X₇R-antagonist and P2X₇R-deficient animals, the role of this receptor in acute and chronic experimental asthma was explored. P2X₇R was found to be up-regulated during acute and chronic asthmatic airway inflammation in mice and humans. In vivo experiments revealed the functional relevance of this finding because selective P2X₇R inhibition or P2X₇R deficiency was associated with reduced features of acute and chronic asthma in the ovalbumin-alum or HDM model of AAI. Experiments with bone marrow chimeras emphasized that P2X₇R expression on hematopoietic cells is responsible for the proasthmatic effects of P2X₇R signaling. In the DC-driven model of AAI, P2X₇R-deficient DCs showed a reduced capacity to induce Th2 immunity in vivo. Up-regulation of P2X₇R on BAL macrophages and blood eosinophils could be observed in patients with chronic asthma. Our data suggest that targeting P2X₇R on hematopoietic cells (e.g., DCs or eosinophils) might be a new therapeutic option for the treatment of asthma.
American Journal of Respiratory Cell and Molecular Biology 04/2011; 44(4):456-64. · 5.13 Impact Factor
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Monica Lucattelli,
Sanja Cicko,
Tobias Müller,
Marek Lommatzsch,
Giovanna De Cunto,
Silvia Cardini,
William Sundas,
Melanie Grimm,
Robert Zeiser,
Thorsten Dürk,
Gernot Zissel, Stephan Sorichter,
Davide Ferrari,
Francesco Di Virgilio,
J Christian Virchow,
Giuseppe Lungarella,
Marco Idzko
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ABSTRACT: Extracellular ATP is up-regulated in the airways of patients with chronic obstructive pulmonary disease, and may contribute to the pathogenesis of the disease. However, the precise mechanisms are poorly understood. Our objective was to investigate the functional role of the ATP receptor P2X(7) in the pathogenesis of cigarette smoke (CS)-induced lung inflammation and emphysema in vivo. Expression of the P2X(7) receptor (P2X(7)R) was measured in lung tissue und immune cells of mice with CS-induced lung inflammation. In a series of experiments using P2X(7) antagonists and genetically engineered mice, the functional role of the P2X(7)R in CS-induced lung inflammation was explored. CS-induced inflammation was associated with an up-regulation of the P2X(7)R on blood and airway neutrophils, alveolar macrophages, and in whole lung tissue. Selective intrapulmonary inhibition of the P2X(7)R reduced CS-induced lung inflammation and prevented the development of emphysema. Accordingly, P2X(7)R knockout mice showed a reduced pulmonary inflammation after acute CS exposure. Experiments with P2X(7)R chimera animals revealed that immune cell P2X(7)R expression plays an important role in CS-induced lung inflammation and emphysema. Extracellular ATP contributes to the development of CS-induced lung inflammation and emphysema via activation of the P2X(7)R. Inhibition of this receptor may be a new therapeutic target for the treatment of chronic obstructive pulmonary disease.
American Journal of Respiratory Cell and Molecular Biology 03/2011; 44(3):423-9. · 5.13 Impact Factor
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ABSTRACT: Pulmonary hypertension (PH) is associated with platelet activation, vascular inflammation and endothelial dysfunction leading to often life threatening thrombo-embolic complications. Microparticles (MPs) are cell vesicles with strong coagulatory and inflammatory effects being released during cell activation and apoptosis. As there are currently no established surrogate markers predicting platelet activation and pro-coagulation in PH patients, the aim of the study was to analyze different pro-coagulatory MP populations that might be related to thrombo-embolic complications in PH patients. Circulating MPs from platelet- (PMP, CD31(+)/61(+)), leukocyte- (LMP, CD11b(+)) and endothelial- (EMP, CD62E(+)) origin were measured by flow cytometry in 19 PH patients and were compared to 16 controls. PH patients had increased levels of PMP (PH vs. control 1,016 ± 201 vs. 527 ± 59 counts per min [cpm], P = 0.032), LMP (PH vs. control 31 ± 3 cpm vs. 18 ± 2 cpm, P = 0.001) and EMP (PH vs. control 99 ± 14 cpm vs. 46 ± 6 cpm, P = 0.001). Furthermore, PMP correlated to LMP (PMP vs. LMP: r = 0.75, P < 0.001) and LMP correlated to EMP levels (LMP vs. EMP, r = 0.74, P < 0.001) indicating a functional interaction between the different types of MP. In comparison to non-embolic PH patients, patients with a thrombo-embolic PH suffered from enhanced endothelial cell dysfunction as represented by significantly increased EMP levels (thrombo-embolic PH vs. non-embolic PH 137 ± 27 vs. 72 ± 10, P = 0.02). PH patients have increased levels of platelet-, leukocyte- and endothelial MP indicating an increased vascular pro-coagulation and inflammation which might be related to thrombo-embolic complications as well as PH progression.
Journal of Thrombosis and Thrombolysis 02/2011; 31(2):173-9. · 1.48 Impact Factor
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ABSTRACT: Extensive breath-hold (BH) diving imposes high pulmonary stress by performing voluntary lung hyperinflation maneuvers (glossopharyngeal insufflation, GI), hyperinflating the lung up to 50% of total lung capacity. Breath-hold durations of up to 10 min without oxygen support may also presume cerebral alterations of respiratory drive. Little is known about the long-term effects of GI onto the pulmonary parenchyma and respiratory adaptation processes in this popular extreme sport.
Lung function assessments and subsequent measures of pulmonary static compliance were obtained for 5 min after GI in 12 elite competitive breath-hold divers (BHD) with a mean apnea diving performance of 6.6 yr. Three-year follow-up measurements were performed in 4 BHD. Respiratory drive was assessed in steady-state measurements for 6% and 9% CO2 in ambient air.
Short-term pulmonary stress effects for static compliance during GI (13.75 L·kPa) could be confirmed in these 12 divers without exhibiting permanent changes to the lungs' distensibility (7.41 L·kPa) or lung function parameters as confirmed by the follow-up measurements and for 4 BHD after 3 yr (P>0.05). Respiratory drive was significantly reduced in these BHD revealing a characteristic breathing pattern with a significant increase in VE and mouth occlusion pressure (P0.1) between free breathing and 6% CO2, as well as between 6% CO2 and 9% CO2 (all P<0.001).
BH diving with performance of GI does not permanently alter pulmonary distensibility or impair ventilatory flows and volumes. A blunted response to elevated CO2 concentrations could be demonstrated, which was supportive of the hypothesis that CO2 tolerance is a training effect due to BH diving rather than being an inherited phenomenon.
Medicine and science in sports and exercise 12/2010; 43(7):1214-9. · 3.71 Impact Factor
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Konrad Wilhelm,
Jayanthi Ganesan,
Tobias Müller,
Christoph Dürr,
Melanie Grimm,
Andreas Beilhack,
Christine D Krempl, Stephan Sorichter,
Ulrike V Gerlach,
Eva Jüttner,
Alf Zerweck,
Frank Gärtner,
Patrizia Pellegatti,
Francesco Di Virgilio,
Davide Ferrari,
Neeraja Kambham,
Paul Fisch,
Jürgen Finke,
Marco Idzko,
Robert Zeiser
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ABSTRACT: Danger signals released upon cell damage can cause excessive immune-mediated tissue destruction such as that found in acute graft-versus-host disease (GVHD), allograft rejection and systemic inflammatory response syndrome. Given that ATP is found in small concentrations in the extracellular space under physiological conditions, and its receptor P2X(7)R is expressed on several immune cell types, ATP could function as a danger signal when released from dying cells. We observed increased ATP concentrations in the peritoneal fluid after total body irradiation, and during the development of GVHD in mice and in humans. Stimulation of antigen-presenting cells (APCs) with ATP led to increased expression of CD80 and CD86 in vitro and in vivo and actuated a cascade of proinflammatory events, including signal transducer and activator of transcription-1 (STAT1) phosphorylation, interferon-γ (IFN-γ) production and donor T cell expansion, whereas regulatory T cell numbers were reduced. P2X(7)R expression increased when GVHD evolved, rendering APCs more responsive to the detrimental effects of ATP, thereby providing positive feedback signals. ATP neutralization, early P2X(7)R blockade or genetic deficiency of P2X(7)R during GVHD development improved survival without immune paralysis. These data have major implications for transplantation medicine, as pharmacological interference with danger signals that act via P2X(7)R could lead to the development of tolerance without the need for intensive immunosuppression.
Nature medicine 12/2010; 16(12):1434-8. · 27.14 Impact Factor
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Sanja Cicko,
Monica Lucattelli,
Tobias Müller,
Marek Lommatzsch,
Giovanna De Cunto,
Silvia Cardini,
William Sundas,
Melanine Grimm,
Robert Zeiser,
Thorsten Dürk,
Gernot Zissel,
Jean-Marie Boeynaems, Stephan Sorichter,
Davide Ferrari,
Francesco Di Virgilio,
J Christian Virchow,
Giuseppe Lungarella,
Marco Idzko
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ABSTRACT: Extracellular ATP acts as a "danger signal" and can induce inflammation by binding to purinergic receptors. Chronic obstructive pulmonary disease is one of the most common inflammatory diseases associated with cigarette smoke inhalation, but the underlying mechanisms are incompletely understood. In this study, we show that endogenous pulmonary ATP levels are increased in a mouse model of smoke-induced acute lung inflammation and emphysema. ATP neutralization or nonspecific P2R-blockade markedly reduced smoke-induced lung inflammation and emphysema. We detected an upregulation the purinergic receptors subtypes on neutrophils (e.g., P2Y2R), macrophages, and lung tissue from animals with smoke-induced lung inflammation. By using P2Y(2)R deficient ((-/-)) animals, we show that ATP induces the recruitment of blood neutrophils to the lungs via P2Y(2)R. Moreover, P2Y(2)R deficient animals had a reduced pulmonary inflammation following acute smoke-exposure. A series of experiments with P2Y(2)R(-/-) and wild type chimera animals revealed that P2Y(2)R expression on hematopoietic cell plays the pivotal role in the observed effect. We demonstrate, for the first time, that endogenous ATP contributes to smoke-induced lung inflammation and then development of emphysema via activation of the purinergic receptor subtypes, such as P2Y(2)R.
The Journal of Immunology 07/2010; 185(1):688-97. · 5.79 Impact Factor
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Marek Lommatzsch,
Sanja Cicko,
Tobias Müller,
Monica Lucattelli,
Kai Bratke,
Paul Stoll,
Melanie Grimm,
Thorsten Dürk,
Gernot Zissel,
Davide Ferrari,
Francesco Di Virgilio, Stephan Sorichter,
Giuseppe Lungarella,
J Christian Virchow,
Marco Idzko
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ABSTRACT: Extracellular ATP promotes inflammation, but its role in chronic obstructive pulmonary disease (COPD) is unknown.
To analyze the expression of ATP and its functional consequences in never-smokers, asymptomatic smokers, and patients with COPD.
ATP was quantified in bronchoalveolar lavage fluid (BALF) of never-smokers, asymptomatic smokers, and patients with COPD of different severity. The expression of specific ATP (purinergic) receptors was measured in airway macrophages and blood neutrophils from control subjects and patients with COPD. The release of mediators by macrophages and neutrophils and neutrophil chemotaxis was assessed after ATP stimulation.
Chronic smokers had elevated ATP concentrations in BALF compared with never-smokers. Acute smoke exposure led to a further increase in endobronchial ATP concentrations. Highest ATP concentrations in BALF were present in smokers and ex-smokers with COPD. In patients with COPD, BALF ATP concentrations correlated negatively with lung function and positively with BALF neutrophil counts. ATP induced a stronger chemotaxis and a stronger elastase release in blood neutrophils from patients with COPD, as compared with control subjects. In addition, airway macrophages from patients with COPD responded with an increased secretion of proinflammatory and tissue-degrading mediators after ATP stimulation. These findings were accompanied by an up-regulation of specific purinergic receptors in blood neutrophils and airway macrophages of patients with COPD.
COPD is characterized by a strong and persistent up-regulation of extracellular ATP in the airways. Extracellular ATP appears to contribute to the pathogenesis of COPD by promoting inflammation and tissue degradation.
American Journal of Respiratory and Critical Care Medicine 05/2010; 181(9):928-34. · 11.08 Impact Factor
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Monika Eichinger,
Stephan Walterspacher,
Tobias Scholz,
Ralf Tetzlaff,
Michael Puderbach,
Kay Tetzlaff,
Annette Kopp-Schneider,
Sebastian Ley,
Kyuok Choe,
Hans-Ulrich Kauczor, Stephan Sorichter
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ABSTRACT: Acute voluntary lung hyperinflation provoked by glossopharyngeal insufflation (GI) elicits numerous, possibly deleterious, effects on the cardiopulmonary system by increasing intrathoracic pressures far above normal values. This study quantifies acute pulmonary hemodynamics during GI using phase-contrast magnetic resonance imaging (MRI).
Hemodynamic parameters were measured in nine elite male breath hold divers with a mean age of 30 yr (range = 20-43 yr) by velocity-encoding cine (VEC)-MRI of the main pulmonary artery (PA) before, during, and after GI. Simultaneously, GI-lung volume (GIVEC-MRI) was measured by MR-compatible spirometry.
Hemodynamic parameters were associated with GIVEC-MRI. Highly significant changes during GI were shown for the mean flow in the PA, which decreased by 45% (P < 0.007), and right ventricular output and cardiac index, which decreased by 41% and 40%, respectively (P < 0.007). Acceleration time also decreased highly significant by 36% during GI (P < 0.007). All hemodynamic parameters except acceleration time returned to baseline after GI.
Acute voluntary lung hyperinflation mimics changes seen in pulmonary arterial hypertension, but unlike the latter, these changes are fully reversible shortly after cessation of voluntary lung hyperinflation. Persistent changes due to repetitive GI could not be detected.
Medicine and science in sports and exercise 02/2010; 42(9):1688-95. · 3.71 Impact Factor
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ABSTRACT: The aim of this study was to investigate the effects of obstructive sleep apnea (OSA) on procedural and declarative memory encoding in the evening prior to sleep, on memory consolidation during subsequent sleep, and on retrieval in the morning after sleep.
Memory performance (procedural mirror-tracing task, declarative visual and verbal memory task) and general neuropsychological performance were assessed before and after one night of polysomnographic monitoring in 15 patients with moderate OSA and 20 age-, sex-, and IQ-matched healthy subjects.
Encoding levels prior to sleep were similar across groups for all tasks. Conventional analyses of averaged mirror tracing performance suggested a significantly reduced overnight improvement in OSA patients. Single trial analyses, however, revealed that this effect was due to significantly flattened learning curves in the evening and morning session in OSA patients. OSA patients showed a significantly lower verbal retention rate and a non-significantly reduced visual retention rate after sleep compared to healthy subjects. Polysomnography revealed a significantly reduced REM density, increased frequency of micro-arousals, elevated apnea-hypopnea index, and subjectively disturbed sleep quality in OSA patients compared to healthy subjects.
The results suggest that moderate OSA is associated with a significant impairment of procedural and verbal declarative memory. Future work is needed to further determine the contribution of structural or functional alterations in brain circuits relevant for memory, and to test whether OSA treatment improves or normalizes the observed deficits in learning.
Journal of clinical sleep medicine: JCSM: official publication of the American Academy of Sleep Medicine 12/2009; 5(6):540-8. · 3.23 Impact Factor
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ABSTRACT: Pneumonia caused by Pseudomonas (P.) aeruginosa is a leading cause of morbidity and mortality in patients with chronic lung diseases. Systemic vaccination in patients with cystic fibrosis has been only successful in part. Mucosal vaccination could lead to enhanced airway immunogenicity. Pathogen specific secretory IgA antibodies could prevent bacterial invasion into the lung mucosa.
A phase 1-2 mucosal vaccination trial with an intranasal P. aeruginosa vaccine was performed.
12 patients with chronic lung diseases (8 COPD, 2 cystic fibrosis, 1 bronchiectasis, 1 histiocytosis X) were vaccinated three times intranasally followed by a systemic booster vaccination with a recombinant hybrid protein encompassing the main protective epitopes of two outer membrane proteins of P. aeruginosa. Mucosal and systemic antibody responses were measured after boosting and after a half-year follow-up compared to a representative control cohort.
Specific IgG and IgA antibodies in the patient's sera, saliva and sputum were determined by enzyme-linked immunosorbent assay (ELISA) and IgG subclass distributions were defined with monoclonal mouse antibodies.
Both forms of vaccination were well tolerated. Significant elevated IgA and IgG antibodies could be measured in sputum, saliva and in the sera of 11/12 patients.
Mucosal vaccination followed by systemic boost with an outer membrane protein vaccine against P. aeruginosa leads to airway immunogenicity against the pathogen. Further clinical trials should elucidate the protective efficacy of this vaccination method.
Vaccine 04/2009; 27(21):2755-9. · 3.77 Impact Factor
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Tobias Müller,
Thorsten Dürk,
Britta Blumenthal,
Melanie Grimm,
Sanja Cicko,
Elisabeth Panther, Stephan Sorichter,
Yared Herouy,
Francesco Di Virgilio,
Davide Ferrari,
Johannes Norgauer,
Marco Idzko
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ABSTRACT: Beside its well described role in the central and peripheral nervous system 5-hydroxytryptamine (5-HT), commonly known as serotonin, is also a potent immuno-modulator. Serotoninergic receptors (5-HTR) are expressed by a broad range of inflammatory cell types, including dendritic cells (DCs). In this study, we aimed to further characterize the immuno-biological properties of serotoninergic receptors on human monocyte-derived DCs. 5-HT was able to induce oriented migration in immature but not in LPS-matured DCs via activation of 5-HTR(1) and 5-HTR(2) receptor subtypes. Accordingly, 5-HT also increased migration of pulmonary DCs to draining lymph nodes in vivo. By binding to 5-HTR(3), 5-HTR(4) and 5-HTR(7) receptors, 5-HT up-regulated production of the pro-inflammatory cytokine IL-6. Additionally, 5-HT influenced chemokine release by human monocyte-derived DCs: production of the potent Th1 chemoattractant IP-10/CXCL10 was inhibited in mature DCs, whereas CCL22/MDC secretion was up-regulated in both immature and mature DCs. Furthermore, DCs matured in the presence of 5-HT switched to a high IL-10 and low IL-12p70 secreting phenotype. Consistently, 5-HT favoured the outcome of a Th2 immune response both in vitro and in vivo. In summary, our study shows that 5-HT is a potent regulator of human dendritic cell function, and that targeting serotoninergic receptors might be a promising approach for the treatment of inflammatory disorders.
PLoS ONE 02/2009; 4(7):e6453. · 4.09 Impact Factor
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ABSTRACT: Extracellular nucleotides are the focus of increasing attention for their role as extracellular mediators since they are released into the extracellular environment in a regulated manner and/or as a consequence of cell damage.Here, we show that human eosinophils stimulated with different nucleotides release eosinophil cationic protein (ECP) and the chemokine interleukin 8 (IL-8), and that release of these two proteins has a different nucleotide requirement.Release of ECP was triggered in a dose-dependent manner by ATP, UTP and UDP, but not by 2′-&3′-o-(4-benzoyl-benzoyl)adenosine 5′-triphosphate (BzATP), ADP and α,β-methylene adenosine 5′ triphosphate (α,β-meATP). Release of IL-8 was triggered by UDP, ATP, α,β-meATP and BzATP, but not by UTP or ADP. Pretreatment with pertussis toxin abrogated nucleotide-stimulated ECP but not IL-8 release.Release of IL-8 stimulated by BzATP was fully blocked by the P2X7 blocker KN-62, while release triggered by ATP was only partially inhibited. IL-8 secretion due to UDP was fully insensitive to KN-62 inhibition.Priming of eosinophils with GM-CSF increased IL-8 secretion irrespectively of the nucleotide used as a stimulant.It is concluded that extracellular nucleotides trigger secretion of ECP by stimulating a receptor of the P2Y subfamily (possibly P2Y2), while, on the contrary, nucleotide-stimulated secretion of IL-8 can be due to activation of both P2Y (P2Y6) and P2X (P2X1 and P2X7) receptors.British Journal of Pharmacology (2003) 138, 1244–1250. doi:10.1038/sj.bjp.0705145
British Journal of Pharmacology 01/2009; 138(7):1244 - 1250. · 4.41 Impact Factor
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Daniel Myrtek,
Tobias Müller,
Verena Geyer,
Natalie Derr,
Davide Ferrari,
Gernot Zissel,
Thorsten Dürk, Stephan Sorichter,
Werner Luttmann,
Michael Kuepper,
Johannes Norgauer,
Francesco Di Virgilio,
J Christian Virchow,
Marco Idzko
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ABSTRACT: Alveolar macrophages play a crucial role in the pathogenesis of inflammatory airway diseases. By the generation and release of different inflammatory mediators they contribute to both recruitment of different leukocytes into the lung and to airway remodeling. A potent stimulus for the release of inflammatory cytokines is ATP, which mediates its cellular effects through the interaction with different membrane receptors, belonging to the P2X and P2Y families. The aim of this study was to characterize the biological properties of purinoceptors in human alveolar macrophages obtained from bronchoalveolar lavages in the context of inflammatory airway diseases. The present study is the first showing that human alveolar macrophages express mRNA for different P2 subtypes, namely P2X(1), P2X(4), P2X(5), P2X(7), P2Y(1), P2Y(2), P2Y(4), P2Y(6), P2Y(11), P2Y(13), and P2Y(14). We also showed that extracellular ATP induced Ca(2+) transients and increased IL-1beta secretion via P2X receptors. Furthermore, extracellular nucleotides inhibited production of IL-12p40 and TNF-alpha, whereas IL-6 secretion was up-regulated. In summary, our data further support the hypothesis that purinoceptors are involved in the pathogenesis of inflammatory lung diseases.
The Journal of Immunology 08/2008; 181(3):2181-8. · 5.79 Impact Factor
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ABSTRACT: Competitive breath-hold divers (BHD) employ glossopharyngeal insufflation (GI) to increase intrapulmonary oxygen stores and prevent the lungs from dangerous compressions at great depths. Glossopharyngeal insufflation is associated with inflation of the lungs beyond total lung capacity (TLC). It is currently unknown whether GI transiently over-distends the lungs or adversely affects lung elastic properties in the long-term. Resting lung function, ventilatory drive, muscle strength, and lung compliance were measured in eight BHD who performed GI since 5.5 (range 2-6) years on average, eight scuba divers, and eight control subjects. In five BHD subsequent measures of static lung compliance (Cstat) were obtained after 1 and 3 min following GI. Breath-hold divers had higher than predicted ventilatory flows and volumes and did not differ from control groups with regard to gas transfer, inspiratory muscle strength, and lung compliance. A blunted response to CO2 was obtained in BHD as compared to control groups. Upon GI there was an increase in mean vital capacity (VCGI) by 1.75 +/- 0.85 (SD) L compared to baseline (p < 0.001). In five BHD Cstat raised from 3.7 (range 2.9-6.8) L/kPa at baseline to 8.1 (range 3.4-21.2) L/kPa after maximal GI and thereafter gradually decreased to 5.6 (range 3.3-8.1) L/kPa after 1 min and 4.2 (range 2.7-6.6) L/kPa after 3 min (p < 0.01). We conclude that in experienced BHD there is a transient alteration in lung elastic recoil. Resting lung function did not reveal a pattern indicative of altered lung ventilatory or muscle function.
Arbeitsphysiologie 04/2008; 103(4):469-75. · 2.15 Impact Factor
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ABSTRACT: There is accumulating evidence that points to a role of serotonin (5-hydroxytryptamine [5-HT]) in the pathophysiology of asthma. Therefore, we analyzed the expression of serotoninergic receptors (5-HTR), its linkage to intracellular calcium homeostasis, and its influence on the production and secretion of IL-6, prostaglandin E(2), the CCL-Chemokine CCL5/Rantes, and the CXC-chemokines CXCL8/IL-8, CXCL9/MIG, CXCL10/IP-10, and CXCL11/I-TAC in primary alveolar epithelial cells type II and the human lung cell lines A549 and BEAS-2B. Employing a PCR approach we were able to demonstrate mRNA expression of several 5-HTR, such as the heptahelical receptors 5-HTR1A, 5-HTR1B, 5-HTR1E, 5-HTR1F, 5-HTR2A, 5-HTR4, 5-HTR6, and 5-HTR7, as well as the ligand-gated ion channel 5-HTR3 in alveolar epithelial cells type II (AEC-II), A549, and BEAS-2B cells. To verify functional expression of 5-HTR subtypes, Ca(2+)-transients were analyzed. This enabled us to show that 5-HT induced an increase in intracellular calcium. Further experiments with isotype-selective receptor agonists allowed us to demonstrate that 5-HT induced calcium transients via activation of 5-HTR1, 5-HTR2, and 5-HTR3 in A549 and BEAS-2B cells. Moreover, we revealed that stimulation of 5-HTR1 and 5-HTR2 induced Ca(2+) mobilization from intracellular stores, whereas activation of 5-HTR3 induced Ca(2+) influx from the extracellular space. Functional studies indicated that activation of 5-HTR1B, 5-HTR1E/F, 5-HTR2, 5-HTR3, 5-HTR4, and 5-HTR7 regulated the release of the cytokine IL-6 and the CXC-chemokine CXCL8/IL-8. Our study shows that 5-HT stimulates different signaling pathways and regulates cytokine release in airway epithelial cells. In summary, our data implicate a pathophysiologic role of 5-HT in the asthmatic inflammatory responses in human airway epithelial cells.
American Journal of Respiratory Cell and Molecular Biology 02/2007; 36(1):85-93. · 5.13 Impact Factor
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ABSTRACT: Dyspnea and fatigue are frequent but poorly understood symptoms in sarcoidosis patients. This study was aimed at assessing the clinical impact of inspiratory muscle impairment on dyspnea and exercise tolerance. This is the first study using nonvolitional tests that are independent of the patient's cooperation and motivation in addition to volitional tests of inspiratory muscle strength in patients with sarcoidosis.
Peak maximal inspiratory mouth pressure (Pimaxpeak), maximal inspiratory pressure sustained for 1.0 s (Pimax1.0), twitch mouth pressure (TwPmo), lung function test results, blood gas measurements, 6-min walking distance (6MWD), and Borg dyspnea scale (BDS) scores were assessed in 24 male sarcoidosis patients and 24 healthy male control subjects matched for age and body mass index.
Mean (+/- SD) Pimaxpeak (95.2 +/- 25.3% vs 124.6 +/- 23.4% predicted, respectively; p < 0.001) and Pimax1.0 (85.6 +/- 31.4% vs 125.8 +/- 26.8% predicted, respectively; p < 0.001) were lower in sarcoidosis patients compared to control subjects. TwPmo tended to be lower in sarcoidosis patients, and there were three patients who had TwPmo values of < 1.0 kPa, which is a strong indicator of inspiratory muscle weakness. The mean 6MWD was 582 +/- 97 m in sarcoidosis patients and 638 +/- 65 in control subjects (p = 0.025). The mean BDS score was higher in sarcoidosis patients (3.3 +/- 1.7 vs 0.2 +/- 0.5, respectively; p < 0.001). Compared to maximal inspiratory pressure, lung function parameters, and blood gas levels, TwPmo was the strongest predictor for 6MWD (r = 0.663; p = 0.003) and BDS score (r = 0.575; p = 0.012) in sarcoidosis patients following multiple linear regression analysis.
Impairment of inspiratory muscle strength occurs in sarcoidosis patients, and has been suggested to be an important factor causing dyspnea and reduced walking capacity, but this is only reliably detectable when using nonvolitional tests of inspiratory muscle strength.
Chest 12/2006; 130(5):1496-502. · 5.25 Impact Factor
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ABSTRACT: Histamine is a well-known mediator eliciting a broad range of responses in different cell types. Four different subtypes of G protein-coupled histamine receptors (H1-H4) have been cloned and pharmacologically characterized. However, involvement of the different histamine receptor subtypes in immunomodulatory functions of bronchial epithelium has only been investigated marginally. The expression and function of histamine receptor subtypes on the human bronchial epithelial cell line BEAS-2B was analyzed by PCR, intracellular Ca++ -measurements and ELISA. We show mRNA expression of the histamine receptor subtypes H1, H2, and H3, but not H4 in the human bronchial epithelial cell line BEAS-2B. Using intracellular Ca++ -measurements, we demonstrated functional expression of the H1 and H3 receptors. To characterize the biological properties of histamine in airway epithelial biology, we also investigated its effects on cytokine secretion by BEAS-2B cells. Thereby, we were able to show up-regulation of the proinflammatory mediators IL-6 and CXCL8/ IL-8 via activation of the H1, H2 and H3 receptor subtypes. The Th1 cytokines CXCL9/MIG and CXCL10/IP-10 and the chemokine CCL5/RANTES were regulated in a distinct manner: Whereas histamine inhibited the IFN-gamma/TNF-alpha-induced secretion of MIG via the histamine receptor subtypes H1, H2, and H3, the histamine-induced suppression of RANTES was due to activation of the H2 and H3 receptors, while reduction of cytokine-triggered IP-10 secretion was mediated only by triggering the H2 receptor. In summary our data provide evidence that histamine released during allergic lung diseases exerts regulatory influence on airway epithelial cells.
International Journal of Molecular Medicine 12/2006; 18(5):925-31. · 1.98 Impact Factor
