Growing up in an alpine farming environment with frequent contact to cowsheds is protective against allergic diseases. Arabinogalactan from this environment is one of the protective compounds. The source of this polysaccharide was traced back to grasses such as Alopecurus pratensis. This study shows that the total synthesis of arabinose and galactose containing partial structures from such arabinogalactan is possible. After coupling of these pentasaccharides to a carrier protein, the glycoconjugates protect against allergic response in a mouse model asthma. More information can be found in the Communication by T. Opatz, M. Peters, et al. on page 928.

Arabinogalactan, a microheterogeneous polysaccharide occurring in plants, is known for its allergy-protective activity, which could potentially be used for preventive allergy treatment. New treatment options are highly desirable, especially in a preventive manner, due to the constant rise of atopic diseases worldwide. The structural origin of the allergy-protective activity of arabinogalactan is, however, still unclear and isolation of the polysaccharide is not feasible for pharmaceutical applications due to a variation of the activity of the natural product and contaminations with endotoxins. Therefore, a pentasaccharide partial structure was selected for total synthesis and subsequently coupled to a carrier protein to form a neoglycoconjugate. The allergy-protective activity of arabinogalactan could be reproduced with the partial structure in subsequent in vivo experiments. This is the first example of a successful simplification of arabinogalactan with a single partial structure while retaining its allergy-preventive potential.

BACKGROUND:Neuropeptide S Receptor 1 ( NPSR1) and Retinoid Acid Receptor-Related Orphan Receptor Alpha (RORA ) interact biologically, are both known candidate genes for asthma, and are involved in controlling circadian rhythm. Thus, we assessed (1) whether interactions between RORA and NPSR1 specifically affect the nocturnal asthma phenotype and (2) how this may differ from other asthma phenotypes. METHODS:Interaction effects between 24 single-nucleotide polymorphisms (SNPs) in RORA and 35 SNPs in NPSR1 on asthma and nocturnal asthma symptoms were determined in 1432 subjects (763 asthmatics [192 with nocturnal asthma symptoms]; 669 controls) from the Multicenter Asthma Genetic in Childhood/International Study of Asthma and Allergies in Childhood studies. The results were validated and extended in children from the Manchester Asthma and Allergy Study (N = 723) and the Children Allergy Milieu Stockholm and Epidemiological cohort (N = 1646). RESULTS:RORA* NPSR1 interactions seemed to affect both asthma and nocturnal asthma. In stratified analyses, however, interactions mainly affected nocturnal asthma and less so asthma without nocturnal symptoms or asthma severity. Results were replicated in two independent cohorts and seemed to remain constant over time throughout youth. CONCLUSION:RORA* NPSR1 interactions appear to be involved in mechanisms specific for nocturnal asthma. In contrast to previous studies focusing on the role of beta 2 receptor polymorphisms in nocturnal asthma as a feature of asthma control or severity in general, our data suggest that changes in circadian rhythm control are associated with nighttime asthma symptoms.

Influenza A, rhinovirus and the human respiratory syncytial virus (RSV) are leading causes of severe lower respiratory tract infections in young children, immunocompromised patients and the elderly. Up to 50% of the children under the age of 16 with these viral infections represent with bacterial co-infections. There are hints from the literature that these bacterial co-infections may aggravate the clinical symptoms substantially, while the reason for this observation especially in premature babies is still unknown. In search for a clinical relevant animal model of RSV-infection we compared BALB/c with DBA/2 N mice during infection, the later suffering from a complement factor 5 (C5) deficiency. It turned out that DBA/2 N mice, especially when co-infected with mycoplasma spp, showed significantly more clinical symptoms, higher viral replication in lung homogenate, strong influx of inflammatory cells and increased airway hyperreactivity (AHR). This data was confirmed by co-infection of RSV and mycoplasma spp. in a C5a-receptor knock-out (C5aR1-KO) also showing enhanced clinical signs of infection like weight loss, prominent invasion of inflammatory cells into the lung and AHR. The connection between infection based AHR and induction of cysteinyl leukotrienes (CysLT) by viral pathogens is known; interestingly, we observed that co-infection in C5aR1-KO-mice led to higher levels of CysLT, which in turn resulted in a more distinctive AHR. Cell influx into BALF and AHR could be reduced by treatment with Zileuton inhibiting the synthesis of CysLT by the 5-Lipoxygenase. Furthermore, the application of recombinant C5a dampened the inflammatory effect after co-infection. In summary our data show a possible link between signaling through the C5aR and the regulation of the synthesis of CysLT. Since CysLT was already recognized as important targets in inflammatory airway disease the modulation of this signaling axis may be another meaningful concept to treat symptoms of severe airway infections.

According to the World Health Organization millions of people worldwide suffer from respiratory diseases such as bronchial Asthma, Ideopathic Pulmonary Fibrosis (IPF) and Chronic Obstructive Pulmonary Disease (COPD) with COPD being the fourth leading cause of death. A persistent inflammatory response has been thought to be the reason for such pulmonary diseases to become chronic. The consequence is a remodeling of the airway tissue structure, where most pulmonary diseases share similar features like epithelial loss, smooth muscle cell (SMC) hyperplasia and hypertrophy, angiogenesis and a deposition of extracellular matrix (fibrosis). Inhaled bronchodilators and corticosteroids only help to decrease the severity of symptoms, but there is no strategy available known to prevent airway remodeling. Nitric oxide (NO), which is excessively generated under allergic airway inflammation, is known to modulate immune responses. In general NO acts via its receptor NO-sensitive guanylyl cyclase (NO-GC), thereby promoting cyclic guanosine monophosphate (cGMP) formation. Consequently, a modulatory role of NO-GC for immune responses is suggested. Furthermore, antifibrotic effects of cGMP by an inhibition of TGFβ-mediated myofibroblast activation have been described. To study the role of NO-GC1 in allergic inflammation and tissue remodeling we used NO-GC1 knockout mice (NO-GC1 k. o.) and sensitized and challenged them with ovalbumin. However, we found reduced allergic inflammation and Th2 response in these mice due to a shift in T-helper cells towards a Th1 phenotype. We suggest a reduced activity of phosphodiesterases in NO-GC k. o. mice, due to an increased intracellular cyclic adenosine monophosphate (cAMP) concentration in dendritic cells, to influence the Th1/Th2 balance. Despite the reduced allergic inflammation we found increased airway fibrosis in the k. o. mice as measured in lung sections stained for collagen. In order to identify the cell type responsible for the elevated tissue remodeling without influencing the immune response, we aim to specifically delete NO-GC1 in SMC, where NOGC-1 is predominantly expressed in the lungs. With the help of a tamoxifen-inducible Cre recombinase system we are able to delete NO-GC 1 specifically in αSMA-expressing cells (SMC, myofibroblasts) of adult mice. This way, the potential exacerbation of tissue remodeling due to a decreased cGMP forming capacity of SMCs and myofibroblasts can be investigated for different pulmonary disease models like chronic OVA-asthma and Bleomycin-induced IPF.

Since the first description of dendritic cells by Steinman and Cohn in 1973, this important cell type has gained increasing attention. Over 4000 papers have been published on this topic annually during the last few years. At the beginning, dendritic cells were recognized for their immune stimulatory properties and their importance in initiating an adaptive immune response. Later, it was found that dendritic cells do not only initiate but also regulate immune responses. This attribute makes the so-called regulatory dendritic cells highly important for the prevention of exaggerated immune responses. Immune cells make contact with different Ags every day and must be tightly controlled to prevent excessive inflammation and subsequent organ destruction, particularly in organs such as the gut and lungs. Here, we give a brief overview of our current knowledge on how immune responses are controlled by dendritic cells, highlighting how they are involved in the induction of peripheral tolerance. We focus on what is known about these processes in the lung, with a closer look at their role in the induction and control of diseases such as bronchial asthma, chronic obstructive pulmonary disease and lung infections. Finally, we summarize some current approaches to modulate the behavior of dendritic cells that may hopefully lead to future therapeutics to control exaggerated immune responses.

BackgroundAllergen-specific immunotherapy is the only causal form of therapy for IgE-mediated allergic diseases. Subcutaneous immunotherapy (SCIT) is considered safe and well tolerated in adults, yet there is less evidence of safety in the pediatric population. MethodsA non-interventional prospective observing longitudinal study was carried out to determine the incidence of local and systemic side effects by SCIT, routinely performed in pediatric patients. A total of 581 pediatric patients were observed in 18 study centers between March 2012 and October 2014, recording 8640 treatments and 10015 injections. ResultsA total of 54.6% of the patients experienced immediate local side effects at least once; delayed local side effects were seen in 56.1%. Immediate systemic adverse reactions occurred in 2.2% of patients; 7.4% experienced delayed systemic side effects. However, severe systemic side effects (grade III in the classification of Ring and Messmer) were seen in 0.03% of all treatments, all appearing within 30minutes after the injections. No grade IV reactions were observed. In addition, many potential risk factors were investigated, yet only a few were found to be associated with the occurrence of side effects. ConclusionsSubcutaneous immunotherapy is a safe form of therapy in pediatric patients, with similar rates of local side effects compared to adult patients and low rates of severe systemic side effects. However, local and systemic reactions occurring later than 30minutes after injection were observed more often than expected, which makes it essential to be attentive on behalf of pediatricians, patients, and parents.

Background: Large sample sizes are needed for sublingual immunotherapy (SLIT) trials because of inherent data variability secondary to inconsistent allergen exposure. Obtaining large sample sizes for pediatric SLIT trials is challenging, but a Bayesian approach using prior adult data can reduce the necessary sample size. Objective: We sought to describe how a Bayesian framework using prior information from adult trials can be used to improve pediatric SLIT clinical development. Methods: Data were compiled by using a frequentist approach (conventional clinical trial approach independent of prior data) from trials conducted during the clinical development of timothy grass SLIT-tablets. Results: The treatment effect of timothy grass SLIT-tablets was considered similar between pediatric (n = 795) and adult (n = 2299) data pools, with relative total combined symptom plus medication score improvement versus placebo of 21% (95% CI, 11.0% to 30.4%) and 20% (95% CI, 14.6% to 24.4%), respectively. Phleum pratense-specific IgG(4) and IgE-blocking factor increased from baseline in both children and adults treated with timothy grass SLIT-tablets. Given the reasonable assumption in similarity of treatment response between adults and children, a Bayesian approach is described to demonstrate rigorous efficacy criteria for pediatric trials incorporating information from prior adult trials and thereby reduce the sample size. Conclusions: Data support the similarity of efficacy and immunologic changes between children and adults treated with SLIT for allergic rhinoconjunctivitis. Therefore it is appropriate to use data from adult trials to design feasible trials in children, which might reduce unsafe off-label use by promoting more quickly proper labeling of approved products.

Allergic airway inflammation is accompanied by excessive generation of nitric oxide (NO). Beside its detrimental activity due to the generation of reactive nitrogen species, NO was found to modulate immune responses by activating the NO-sensitive Guanylyl Cyclases (NO-GCs) thereby mediating the formation of the second messenger cyclic GMP (cGMP). To investigate the contribution of the key-enzyme NO-GC on the development of Th2 immunity in vivo, we sensitized knock-out (KO) mice of the major isoform NO-GC1 to the model allergen ovalbumin (OVA). The loss of NO-GC1 attenuates the Th2 response leading to a reduction of airway inflammation and IgE production. Further, in vitro-generated OVA-presenting DCs of the KO induce only a weak Th2 response in the WT recipient mice upon re-exposure to OVA. In vitro, these NO-GC1 KO BMDCs develop a Th1-polarizing phenotype and display increased cyclic AMP (cAMP) formation, which is known to induce Th1-bias. According to our hypothesis of a NO-GC1/cGMP-dependent regulation of cAMP-levels we further demonstrate activity of the cGMP-activated cAMP-degrading phosphodiesterase 2 in DCs. Herewith, we show that activity of NO-GC1 in DCs is important for the magnitude and bias of the Th response in allergic airway disease most likely by counteracting intracellular cAMP.

Kinder, die auf Bauernhöfen groß werden, sind meistens vor allergischer Sensibilisierung und der damit verbundenen Entstehung von Asthma geschützt. Wir konnten in vorhergehenden Studien zeigen, dass Arabinogalactane (AG), die aus Stallstaubextrakt isoliert werden können, in Mausmodellen für Asthma eine Allergie-protektive Wirkung zeigen. Ziel der hier vorgestellten Studie war es Oberflächenepitope im AG zu identifizieren, die ebenfalls eine Allergie-protektive Wirkung aufweisen.

Die von der Gesellschaft für Hygiene, Umweltmedizin und Präventivmedizin (GHUP) federführend erstellte Leitlinie „Medizinisch klinische Diagnostik bei Schimmelpilzexposition in Innenräumen“ ist Gegenstand des vorliegenden Abschnitts. Schimmelpilzwachstum im Innenraum ist als ein potenzielles Gesundheitsrisiko zu betrachten, auch ohne dass ein quantitativer und/oder kausaler Zusammenhang zwischen dem Vorkommen einzelner Arten und Gesundheitsbeschwerden gesichert werden kann. Abgesehen von der Allergischen Bronchopulmonalen Aspergillose (ABPA) und den durch Schimmelpilze kausal verursachten Mykosen, liegen lediglich ausreichende Evidenzen für folgende Assoziationen von Feuchte-/Schimmelpilzschäden und unterschiedlichen Gesundheitseffekten vor: allergische Atemwegserkrankungen, Asthma (Manifestation, Progression, Exazerbation), allergische Rhinitis, Exogen Allergische Alveolitis, Begünstigung von Atemwegsinfekten/Bronchitis. Dabei ist das sensibilisierende Potenzial von Schimmelpilzen im Vergleich zu anderen Umweltallergenen deutlich geringer einzuschätzen. Aktuelle Studien zeigen europaweit eine vergleichsweise geringe Sensibilisierungsprävalenz von 3–10 % gemessen an der Gesamtbevölkerung. Eingeschränkte oder vermutete Evidenz für eine Assoziation liegt vor hinsichtlich „mucous membrane irritation“ und Atopischen Ekzems (Manifestation, Progression, Exazerbation). Inadäquate oder unzureichende Evidenz für eine Assoziation liegt vor für COPD, akute idiopathische pulmonale Hämorrhagie bei Kindern, Rheuma/Arthritis, Sarkoidose und Krebserkrankungen. Das Infektionsrisiko von den in Innenräumen regelmäßig vorkommenden Schimmelpilzarten ist für gesunde Personen gering, die meisten Arten sind in die Risikogruppe 1 und wenige in 2 (Aspergillus fumigatus, A. flavus) der Biostoffverordnung eingestuft. Nur Schimmelpilze, die potenziell in der Lage sind, Toxine zu bilden, kommen als Auslöser einer Intoxikation in Betracht. Ob im Einzelfall eine Toxinbildung im Innenraum stattfindet, entscheiden die Umgebungs- und Wachstumsbedingungen und hier vor allem das Substrat. Von Geruchswirkungen und/oder Befindlichkeitsstörungen kann bei Feuchte-/Schimmelpilzschäden im Innenraum grundsätzlich jeder betroffen sein. Hierbei handelt es sich nicht um eine Gesundheitsgefährdung. Prädisponierende Faktoren für Geruchswirkungen können genetische und hormonelle Einflüsse, Prägung, Kontext und Adaptationseffekte sein. Prädisponierende Faktoren für Befindlichkeitsstörungen können Umweltbesorgnisse, -ängste, -konditionierungen und -attributionen sowie eine Vielzahl von Erkrankungen sein. Besonders zu schützende Risikogruppen bezüglich eines Infektionsrisikos sind Personen unter Immunsuppression nach der Einteilung der Kommission für Krankenhaushygiene und Infektionsprävention (KRINKO) beim Robert Koch-Institut (RKI) und Personen mit Mukoviszidose (Zystischer Fibrose), bezüglich eines allergischen Risikos Personen mit Mukoviszidose (Zystischer Fibrose) und Personen mit Asthma bronchiale. Die rationale Diagnostik beinhaltet die Anamnese, eine körperliche Untersuchung, eine konventionelle Allergiediagnostik einschließlich gegebenenfalls Provokationstests, vereinzelt sind auch zelluläre Testsysteme indiziert. Zum Vorgehen bei Schimmelpilzinfektionen wird auf die angemeldete AWMF-Leitlinie Diagnose und Therapie invasiver Aspergillus-Infektionen verwiesen. Hinsichtlich Mykotoxine existieren zurzeit keine brauchbaren und validierten Testverfahren, die in der klinischen Diagnostik eingesetzt werden könnten. Präventivmedizinisch ist wichtig, dass Schimmelpilzbefall in relevantem Ausmaß aus Vorsorgegründen nicht toleriert werden darf. Zur Beurteilung des Schadensausmaßes und zum Vorgehen wird auf den „Schimmelpilzleitfaden“ des Umweltbundesamtes verwiesen.

The availability of convenient assays for the detection and quantification of pathogen-associated molecular patterns (PAMPs) is limited. In the case of lipopolysaccharide (LPS) the so-called LAL (limulus amebocyte lysate) test is available, an assay that is performed with the lysate of the blood of the horse shoe crab. Although a sensitive and convenient assay, it lacks specificity, since it is affected by other endotoxins like, for instance, fungal cell walls as well. Here, we describe a bioassay that can be used to detect and quantitate PAMPs in environmental samples. More specific we demonstrate the usage of TLR2 and TLR4/CD14/MD2 transfected Hek293 cells to quantitatively determine bacterial lipoproteins and LPS, respectively. We show the usefulness of these assays to measure LPS in tobacco before and after combustion.