Volatile Organic Compounds Enhance Allergic Airway Inflammation in an Experimental Mouse Model

<div><h3>Background</h3><p>Epidemiological studies suggest an association between exposure to volatile organic compounds (VOCs) and adverse allergic and respiratory symptoms. However, whether VOCs exhibit a causal role as adjuvants in asthma development remains unclear.</p> <h3>Methods</h3><p>To investigate the effect of VOC exposure on the development of allergic airway inflammation Balb/c mice were exposed to VOCs emitted by new polyvinylchloride (PVC) flooring, sensitized with ovalbumin (OVA) and characterized in acute and chronic murine asthma models. Furthermore, prevalent evaporated VOCs were analyzed and mice were exposed to selected single VOCs.</p> <h3>Results</h3><p>Exposure of mice to PVC flooring increased eosinophilic lung inflammation and OVA-specific IgE serum levels compared to un-exposed control mice. The increased inflammation was associated with elevated levels of Th2-cytokines. Long-term exposure to PVC flooring exacerbated chronic airway inflammation. VOCs with the highest concentrations emitted by new PVC flooring were N-methyl-2-pyrrolidone (NMP) and 2,2,4-trimethyl-1,3-pentanediol diisobutyrate (TXIB). Exposure to NMP or TXIB also increased the allergic immune response in OVA-sensitized mice. <em>In vitro</em> or <em>in vivo</em> exposure to NMP or TXIB reduced IL-12 production in maturing dendritic cells (DCs) and enhanced airway inflammation after adoptive DC transfer into Balb/c mice. At higher concentrations both VOCs induced oxidative stress demonstrated by increased isoprostane and glutathione-S-transferase-pi1 protein levels in the lung of non-sensitized mice. Treatment of PVC flooring-exposed mice with N-acetylcysteine prevented the VOC-induced increase of airway inflammation.</p> <h3>Conclusions</h3><p>Our results demonstrate that exposure to VOCs may increase the allergic immune response by interfering with DC function and by inducing oxidative stress and has therefore to be considerate as risk factor for the development of allergic diseases.</p> </div

Additional file 6: Figure S5. of 2.7 million samples genotyped for HLA by next generation sequencing: lessons learned

Mean primer dimer rates of HLA loci for different workflows. Error bars show standard deviation. (PDF 5 kb

Regional HLA Differences in Poland and Their Effect on Stem Cell Donor Registry Planning

<div><p>Regional HLA frequency differences are of potential relevance for the optimization of stem cell donor recruitment. We analyzed a very large sample (<i>n</i> = 123,749) of registered Polish stem cell donors. Donor figures by 1-digit postal code regions ranged from <i>n</i> = 5,243 (region 9) to <i>n</i> = 19,661 (region 8). Simulations based on region-specific haplotype frequencies showed that donor recruitment in regions 0, 2, 3 and 4 (mainly located in the south-eastern part of Poland) resulted in an above-average increase of matching probabilities for Polish patients. Regions 1, 7, 8, 9 (mainly located in the northern part of Poland) showed an opposite behavior. However, HLA frequency differences between regions were generally small. A strong indication for regionally focused donor recruitment efforts can, therefore, not be derived from our analyses. Results of haplotype frequency estimations showed sample size effects even for sizes between <i>n</i>≈5,000 and <i>n</i>≈20,000. This observation deserves further attention as most published haplotype frequency estimations are based on much smaller samples.</p></div

Additional file 1: Figure S1. of 2.7 million samples genotyped for HLA by next generation sequencing: lessons learned

Prevalence of low-quality samples over time and for different sample provenances (DE, Germany; PL, Poland; UK, United Kingdom). (PDF 5 kb

Increased acute allergic immune response by PVC flooring is abrogated by the antioxidant NAC.

<p>Treatment of Balb/c mice with NAC (1g/l) during exposure to PVC flooring reduced the enhanced acute airway inflammation (A), the number of eosinophils in the BAL fluid (B), OVA-specific IgE levels (C), IL-13 and IL-5 cytokine levels (D) and the production of 8-isoprostane in lung homogenates (E) compared to NAC-untreated mice. Data are expressed as mean ± SEM, n ≥ 9 animals per group; *<i>P</i><0.05, PVC flooring <i>vs.</i> OVA; ^#P<0.05, PVC flooring <i>vs</i>. PVC flooring + NAC.</p

NMP and TXIB induce oxidative stress <i>in vitro</i> and <i>in vivo.</i>

<p>A549 human lung epithelial cells were exposed to NMP or TXIB for 24 h. Expression of GSTP-1 was quantified by western blotting (A). Naïve mice were exposed to NMP or TXIB on two consecutive days for 6 hours, lung tissues were taken and GSTP-1 expression was quantified by western blotting (B) and 8-isoprostane (C) was measured as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039817#pone.0039817.s003" target="_blank">File S1</a>. In the pseudoimages, the blue to red coloring represents the lowest to the highest light intensity in NMP (52 µg/m³)- or TXIB (31 µg/m³)-exposed NFκB/luciferase transgenic mice (D). Photon emission was quantified over the lung area (arrow) as mean fold of induction (E). Data are expressed as mean ± SEM from three independent experiments (A), n ≥ 9 animals per group (B, C) or <i>n</i> = 4 per group (D, E); *<i>p</i><0.05 <i>vs</i>. CON.</p

Exposure to PVC flooring increases acute airway inflammation, antigen-specific IgE and Th2 cytokine levels and reduced IFN-γ production in OVA-sensitized Balb/c mice.

<p>Balb/c mice were immunized with OVA (day 1 and 14) and then challenged with OVA on days 14 and 17–19. Mice were exposed to PVC flooring from day 0 to day 20. Effect of PVC flooring on inflammation (H&E, x200, A), verified by an objective, investigator-independent computer-based quantification of lung inflammation (B), on total cell numbers in the BAL fluid (C), OVA-specific IgE levels (D), lung resistance (E), lung compliance (F) and Th2 cytokine levels in the supernatant of OVA-re-stimulated splenocytes (G) or mediastinal lymphnodes (H). Data are expressed as mean ± SEM, n ≥ 9 animals per group; *<i>p</i><0.05 <i>vs</i>. OVA.</p

Long-term exposure to PVC flooring exacerbates chronic airway inflammation.

<p>To induce a chronic asthma phenotype mice were immunized with OVA (day 1 and 14) and then challenged with the antigen twice per week for 8 weeks. Effect of exposure to PVC flooring on inflammation of lung tissues (A), total cell numbers in BAL fluid (B), IgE levels (C), lung resistance (D), dynamic compliance (E), total lung collagen (F), and α-SMA- and PCNA-stained lung sections (G, H). Data are expressed as mean ± SEM, n ≥ 9 animals per group; *<i>P</i><0.05 OVA + PVC <i>vs</i>. OVA; ^#P<0.05 Alum + PVC <i>vs</i>. Alum.</p

VOCs emitted by PVC flooring.

<p>New PVC flooring was installed on the top cover of the mice cage. VOCs emitted by PVC flooring were measured on day 1 as described in the supporting information (<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0039817#pone.0039817.s003" target="_blank">File S1</a>).</p