Andrea Müller

Role of Lipoteichoic Acid in the Phagocyte Response to Group B Streptococcus

Group B Streptococcus (GBS) cell walls potently activate phagocytes by a largely TLR2-independent mechanism. In contrast, the cell wall component lipoteichoic acid (LTA) from diverse Gram-positive bacterial species has been shown to engage TLR2. In this study we examined the role of LTA from GBS in phagocyte activation and the requirements for TLR-LTA interaction. Using cells from knockout mice and genetic complementation in epithelial cells we found that highly pure LTA from both GBS and Staphylococcus aureus interact with TLR2 and TLR6, but not TLR1, in contrast to previous reports. Furthermore, NF-κB activation by LTA required the integrity of two putative PI3K binding domains within TLR2 and was inhibited by wortmannin, indicating an essential role for PI3K in cellular activation by LTA. However, LTA from GBS proved to be a relatively weak stimulus of phagocytes containing ∼20% of the activity observed with LTA from Staphylococcus aureus. Structural analysis by nuclear magnetic resonance spectrometry revealed important differences between LTA from GBS and S. aureus, specifically differences in glycosyl linkage, in the glycolipid anchor and a lack of N-acetylglucosamine substituents of the glycerophosphate backbone. Furthermore, GBS expressing LTA devoid of d-alanine residues, that are essential within immune activation by LTA, exhibited similar inflammatory potency as GBS with alanylated LTA. In conclusion, LTA from GBS is a TLR2/TLR6 ligand that might contribute to secreted GBS activity, but does not contribute significantly to GBS cell wall mediated macrophage activation.

Increased asthma and respiratory symptoms in children exposed to petrochemical pollution.

BACKGROUND Epidemiologic studies show statistical associations between levels of air pollutants and respiratory outcomes. OBJECTIVE We sought to determine the effects of exposure to petrochemical pollution on the respiratory health of children. METHODS Children aged 6 to 12 years living close to the petrochemical plants in La Plata, Argentina (n = 282), were compared with those living in a region with exposure to heavy traffic (n = 270) or in 2 relatively nonpolluted areas (n = 639). Parents answered a validated questionnaire providing health and demographic data. A random sample (n = 181) had lung function measured. Particulate matter and outdoor and indoor volatile organic compound levels were measured during 4-week study periods and reported as overall means for each study area. RESULTS Children living near the petrochemical plant had more asthma (24.8% vs 10.1% to 11.5%), more asthma exacerbations (6.7 vs 2.9-3.6 per year), more respiratory symptoms (current wheeze, dyspnea, nocturnal cough, and rhinitis), and lower lung function (>13% decrease in FEV(1) percent predicted) than those living in other regions. Length of residence in the area was a significant risk factor, but age, sex, body mass index, proximity to busy roads and other nonpetrochemical industries, length of breast-feeding, and socioeconomic and demographic characteristics of children or their families were not. CONCLUSION Exposure to particulate matter and volatile organic compounds arising from petrochemical plants but not from high traffic density was associated ith worse respiratory health in children.

Identification of spore allergens from the indoor mould Aspergillus versicolor

Background:  Indoor mould growth and dampness are associated with respiratory health effects and allergies and several studies demonstrated that mainly Aspergillus versicolor and Penicillium expansum are responsible for indoor mould exposure. In contrast, commercialized test systems to diagnose allergic reactions to this mould species are not available. In this study, allergenic proteins from spores of the indoor relevant species A. versicolor and P. expansum should get detected and identified.

Increased incidence of allergic sensitisation and respiratory diseases due to mould exposure: Results of the Leipzig Allergy Risk children Study (LARS)

To investigate mould effects on health, the concentration of mould spores in air and dust was determined during a prospective cohort study. Clinical outcome was estimated by questionnaires and determination of specific IgE antibodies and intracellular cytokine production of T cells. A significant association was observed between the incidence of respiratory tract infections and exposure to Penicillium spores. Moreover, Aspergillus exposure was found to be associated with allergic rhinitis or related symptoms. In addition, T cells of children exposed to Aspergillus showed a significantly lower content of TH1 cytokines (IFN-gamma, TNF-alpha, IL-2) producing cells. Our data suggest that mould exposure is associated with several effects on health, depending on the species involved.

Epoxide and Thiirane Toxicity In vitro with the Ciliates Tetrahymena pyriformis: Structural Alerts Indicating Excess Toxicity

The 48 h toxicity of 18 organic narcotics, 13 epoxides, and 2 thiiranes toward the ciliates Tetrahymena pyriformis was determined in terms of 50% growth inhibition EC(50). Nominal EC(50) was corrected for volatilization and sorption to quantify the freely dissolved compound fraction in solution. The derived baseline narcosis model served to evaluate toxicity enhancements T(e) as ratios of narcosis-predicted over experimental EC(50) values. Among the nine heterocycles with aliphatic side chains that include two thiiranes, three compounds yielded T(e) > 10, suggesting their covalent binding at nucleophilic protein sites such as -OH, -NHR, and -SH through S(N)2-type ring-opening. As a general trend of this group, T(e) decreases with increasing alkyl group size. Moreover, four of the six nonaliphatic epoxides exerted substantial excess toxicities with T(e) > 10, which could be rationalized by ring-opening activation through negative inductive effect, benzylic stabilization, and phenyl ring H-bonding. By contrast, 1,2 substituted epoxides showed narcosis-level toxicity, despite the opportunity of side-chain Schiff-base formation with protein amino groups. The resulting structural alerts enable an in silico screening of epoxides and thiiranes for their potential to exert excess toxicity. Note that observed differences in T(e) sensitivity between ciliates, bacteria and fish should be taken into account when designing in vitro alternatives to fish toxicity studies.

Spatiotemporal distribution of airborne mould spores in apartments

Indoor air contamination with mould spores currently experiences an increasing interest with respect to their relevance to health. To assess adverse health effects, epidemiological studies combine the health outcome of individuals with their concomitant exposure to airborne spores, which is observed, for example, during the current month. While the latter is representative for the studied period, health effects might also be the result of long term-exposure or emerge in consequence of a peak of pollution throughout the year. To consider such questions, additional information about the spatiotemporal distribution of airborne spores is necessary. This paper aims at elucidating the spatial and temporal variation of spore concentrations in Leipzig, Germany. The analysis is based on 1165 matched pairs of indoor and outdoor measurements taken in the period 1998-2002. All data were collected in the frame of previous epidemiological studies and refer to apartments. The analysis comprised spore concentrations (as CFU m(-3) in air) of the most important genera, such as Penicillium, Aspergillus, Alternaria, Mucorales, Cladosporium, and also for yeasts. We found two groups of fungi differing in their spatiotemporal distribution. As this behaviour can be explained by the predominant origin and growing conditions, we call them indoor-relevant and outdoor-relevant genera. Penicillium species are a representative of the former group, while the latter is well represented by Cladosporium. In the studied period we did not observe a clear trend in the spore concentration. Outdoors there is a year-to-year variation of Cladosporium spore concentrations, which follow the prevalent climatic conditions. For the spore concentration of the outdoor-relevant group a significant annual cycle was observed. Highest concentrations occurred during the summer months and were about 100 x the winter burden. That means, for a direct comparison of measurements of spore concentrations taken during different months the season has to be considered. We summarise the findings in a seasonal model, which is fitted to our measurements. Based on the model we developed a procedure for seasonal adjustment, which enabled us to estimate the annual peak spore concentration utilising one monthly observation.

From the exposome to mechanistic understanding of chemical-induced adverse effects

The exposome encompasses an individuals exposure to exogenous chemicals, as well as endogenous chemicals that are produced or altered in response to external stressors. While the exposome concept has been established for human health, its principles can be extended to include broader ecological issues. The assessment of exposure is tightly interlinked with hazard assessment. Here, we explore if mechanistic understanding of the causal links between exposure and adverse effects on human health and the environment can be improved by integrating the exposome approach with the adverse outcome pathway (AOP) concept that structures and organizes the sequence of biological events from an initial molecular interaction of a chemical with a biological target to an adverse outcome. Complementing exposome research with the AOP concept may facilitate a mechanistic understanding of stress-induced adverse effects, examine the relative contributions from various components of the exposome, determine the primary risk drivers in complex mixtures, and promote an integrative assessment of chemical risks for both human and environmental health.

Different immunomodulatory effects associated with sub-micrometer particles in ambient air from rural, urban and industrial areas

Immunomodulatory effects of chemicals adsorbed to particles with aerodynamic diameter below 0.49 microm (PM(0.5)) collected in winter 2001 at three sampling points (industrial area [LPIn], traffic-influenced urban area [LPCi], and control area [LPCo]) of La Plata, Argentina, were investigated. The sampling of particulate matter was carried out with high-volume collectors using cascade impactors. PM(0.5)-adsorbed compounds were hexane-extracted by accelerated solvent extraction. For immunological investigations, human peripheral blood lymphocytes were activated by phytohemagglutinin and exposed to dimethyl-sulfoxide dilutions of PM(0.5)-extracts for 24h. Vitality/proliferation was quantified using MTT, released interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) by ELISA. Cytokine production but not vitality/proliferation was significantly suppressed by all of the highest extract concentrations. Generally, suppression of IFN-gamma by PM(0.5)-extracts was stronger than those of IL-4. Based on administered mass of PM(0.5), all extracts suppressed IFN-gamma production nearly uniform. Contrary, LPCi-extracts exerted maximum IFN-gamma suppression based either on air volume or regarding PM(0.5)-adsorbed PAH. Also the ranking of PM(0.5)-associated effects on IL-4 production differs in dependence of the chosen reference points, either mass or [microg/ml] or air volume [m(3)/ml] related dust quantities in cell culture. Based on the corresponding air volume, LPCi-extracts inhibited IL-4 production to the maximum extend, whereas suppression of IL-4 was comparable based on concentrations. This indicates that not only the disparate PM(0.5)-masses in air cause varying impacts, but also that disparities in PM(0.5)-adsorbed chemicals provoke different effects on immune responses and shifts in the regulatory balance that might have implications for allergy and cancer development.

Indoor Mould Exposure Reduces Th 1 Reactivity in Early Childhood

A birth cohort study (LARS - Leipzig Allergy Risk Children Study) was used to investigate the influence of indoor mould exposure on T cell function in early childhood. T cell cytokines (IFN-γ, IL-4) were analysed in a sub-group of the LARS cohort (birth: n = 39, 6 months: n = 11, 12 months: n = 50, 36 months: n = 33). Mould burden was assessed by analysis of questionnaires completed at the birth of the child and measurement of indoor mould spores was carried out when the children were 3 years old. Children whose parents reported mould burden or dampness in their dwellings at the time of birth showed lower amounts of IFN-γ producing Th1 cells during the first year of life (significant at 12 months) in comparison to unexposed children. The capacity to produce IL-4 was not altered, however. When the children were 3 years old, an association between measured indoor mould and suppressed Th1 reactivity was observed. The data suggest that indoor mould exposure is associated with reduced Th1 reactivity in early childhood.