The adjuvant effect of particles on allergic immune responses has been shown to increase with decreasing particle size and increasing particle surface area. Like ultrafine particles, carbon nanotubes (CNTs) have nano-sized dimensions and a large relative surface area and might thus increase allergic responses. Therefore, we examined whether single-walled (sw) and multi-walled (mw) CNTs have the capacity to promote allergic responses in mice, first in an sc injection model and thereafter in an intranasal model. Balb/cA mice were exposed to three doses of swCNT, mwCNT, as well as ultrafine carbon black particles (ufCBPs, Printex90) during sensitization with the allergen ovalbumin (OVA). Five days after an OVA booster, OVA-specific IgE, IgG1, and IgG2a antibodies in serum and the numbers of inflammatory cells and cytokine levels in bronchoalveolar lavage fluid (BALF) were determined. Furthermore, ex vivo OVA-induced cytokine release from mediastinal lymph node (MLN) cells was measured. In separate experiments, differential cell counts were determined in BALF 24 h after a single intranasal exposure to the particles in the absence of allergen. We demonstrate that both swCNT and mwCNT together with OVA strongly increased serum levels of OVA-specific IgE, the number of eosinophils in BALF, and the secretion of Th2-associated cytokines in the MLN. On the other hand, only mwCNT and ufCBP with OVA increased IgG2a levels, neutrophil cell numbers, and tumor necrosis factor-alpha and monocyte chemoattractant protein-1 levels in BALF, as well as the acute influx of neutrophils after exposure to the particles alone. This study demonstrates that CNTs promote allergic responses in mice.

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The prevalence of allergic diseases is influenced by sex and age. Although mouse models are widely used in allergy research, few experimental studies have examined the interaction effects of sex and age on allergy outcomes. Our aim was to investigate the individual and combined effects of sex and age on allergic sensitization and inflammation in two mouse models: an intraperitoneal (i.p.) and an intranasal (i.n.) sensitization model. We also investigated how the allergen immunization dose interacted with age and sex in the i.p. model. Female and male mice were immunized i.p. or i.n. with ovalbumin when 1, 6 or 20 weeks old. In both models, allergen challenges were performed by i.n. delivery. Serum antibodies, draining lymph node cytokine release and airway inflammatory responses were assessed. In the i.p. model, the antibody and cytokine levels and airway inflammation were highly influenced by immunization dose and age. The responses increased with age when using a low immunization dose, but decreased with age when using a high immunization dose. In the i.n. model, antibody production and airway tissue inflammation increased with age. Female compared with male mice generally developed more pronounced antibody and inflammatory responses. Relative to older mice, juvenile mice had augmented airway inflammation to allergen exposures. The study demonstrates that immunization dose, sex and age are highly influential on allergy outcomes. To better mimic different life stages of human allergic airway disease, murine models, therefore, require careful optimization.

In the European Union (EU)-funded project Respiratory Allergy and Inflammation due to Ambient Particles (RAIAP), coarse and fine ambient particulate matter (PM) was collected at traffic dominated locations in Oslo, Rome, Lodz, and Amsterdam, in the spring, summer, and winter 2001/2002. PM was also collected in de Zilk, a rural seaside background location in the Netherlands. The aim of this study was to screen the ambient PM fractions for allergy adjuvant activity measured as the production of allergen- (ovalbumin-) specific immunoglobulin (Ig) E following subcutaneous (sc) injection into the footpad of mice. A second aim was to determine whether the 6-d popliteal lymph node (PLN) assay can be used to detect an allergy adjuvant activity. Allergy screening for IgE adjuvant activity showed that in the presence of ovalbumin (Ova) 12 out of 13 of the fine ambient PM fractions exerted a significant IgE adjuvant activity. In contrast, only 3 out of 13 of the coarse PM fractions had significant adjuvant activity. Overall, fine ambient PM exerted significantly greater IgE adjuvant activity per unit mass than coarse PM. No significant differences were observed between locations or seasons. Substantial higher levels of specific components of PM such as vanadium (V), nickel (Ni), zinc (Zn), ammonium (NH(4)), and sulfate (SO(4)) were present in the fine compared to coarse PM fractions. However, differences in the content of these components among fine PM fractions did not reflect the variation in the levels of IgE anti-Ova. Still, when comparing all seasons overall, positive correlations were observed between V, Ni, and SO(4) and the allergen specific IgE levels. The PLN responses (weight and cell number) to Ova and ambient PM in combination were significantly higher than to Ova or PM alone. Still, the PLN assay appears not to be useful as a quantitative assay for screening of allergy adjuvant activity since no correlation was observed between PLN responses and allergen specific IgE levels. In conclusion, fine ambient PM fractions consistently were found to increase the allergen-specific IgE responses more than the coarse ones. Our finding is in agreement with the notion that traffic-related air pollution contributes to the disease burden in asthma and allergy, and points to fine and ultrafine ambient PM as the most important fractions in relation to allergic diseases.

Epidemiological evidence suggesting that exposure to traffic air pollution may enhance sensitization to common allergens in children is increasing, and animal studies support biological plausibility and causality. The effect of air pollution on respiratory symptoms was suggested to be gender dependent. Previous studies showed that allergy-promoting activity of polystyrene particles (PSP) increased with decreasing particle size after footpad injection of mice. The primary aim of this study was to confirm the influence of particle size on the immunoglobulin E (IgE)-promoting capacity of particles in an airway allergy model. A second aim was to examine whether the allergy-promoting capacity of particles was influenced by gender. Female and male mice were intranasally exposed to the allergen ovalbumin (OVA) with or without ultrafine, fine, or coarse PSP modeling the core of ambient air particles. After intranasal booster immunizations with OVA, serum levels of OVA-specific IgE antibodies, and also markers of airway inflammation and cellular responses in the lung-draining mediastinal lymph nodes (MLN), were determined. PSP of all sizes promoted allergic responses, measured as increased serum concentrations of OVA-specific IgE antibodies. Further, PSP produced eosinophilic airway inflammation and elevated MLN cell numbers as well as numerically reducing the percentage of regulatory T cells. Ultrafine PSP produced stronger allergic responses to OVA than fine and coarse PSP. Although PSP enhanced sensitization in both female and male mice, significantly higher IgE levels and numbers of eosinophils were observed in females than males. However, the allergy-promoting effect of PSP was apparently independent of gender. Thus, our data support the notion that ambient air particle pollution may affect development of allergy in both female and male individuals.