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J investing allergol clin immunol 2006 ford

Of the children, children had a waist circumference measurement. There were no significant differences in race, age, or annual household income between children without a waist circumference measurement and those with a waist circumference measurement. There was a borderline difference in gender among children without a waist circumference measurement Outcome definitions Pulmonary function testing was performed and asthma was diagnosed according to the American Thoracic Society guidelines Asthma severity was designated based on lung function FEV1 and frequency of symptoms, and was classified into mild, moderate or severe Allergy skin prick testing SPT Allergic sensitization was determined in all subjects by SPTs to eleven aeroallergens mold mixes 2 , grass mix, ragweed giant and short , tree pollen mixes 2 , weed mix, dust mite mix, cat, dog, and cockroach mix common to the Ohio River Valley Hollister-Stier.

Sensitization was defined as present when there was wheal greater than 3 mm and greater than the saline negative control. A child was considered to have allergic rhinitis if they had rhinitis as well as a positive SPT to at least one aeroallergen.

Spearman correlation coefficients r were used to examine the relationships between BMI percentiles and the other obesity measures. To facilitate the comparison of the obesity measures, the alternative measures were standardized by subtracting the mean and dividing by the standard deviation. All the measures were categorized into three groups based on the 25th and 85th percentile. Since the 95th percentile gave similar trends to the 85th percentile, the 85th percentile was used as the cutoff for the high weight stratum to allow sufficient analytic power.

Allergic rhinitis was chosen as the control group because a large proportion of asthmatics were skin test positive with allergic rhinitis. Among children with allergic rhinitis, the effect of the obesity measures on the probability of mild asthma vs.

Generalized logistic regression models were used to investigate the relationship of the obesity measures with asthma severity in children with allergic rhinitis. Logistic regression was used to investigate the relationship between the obesity measures and the likelihood of having at least one positive SPT versus a negative SPT.

Proportional odds regression was used to determine the association between each obesity measure and the atopy index. The models were tested in the whole population and then stratified by diagnosis asthma vs. Continuous variables were logarithmically transformed if needed to achieve normality. In all models, potential covariates considered include race, age group, gender, maternal and paternal education, smoking in the household, and annual household income.

The Wilcoxon two-sample test with t-approximation was used to compare continuous data that was not normally distributed. Percentile Curves for Obesity Measures Percentile curves were constructed for each obesity measure using quantile regression 36 for children with allergic rhinitis. Fitted values were computed for the 10th, 25th, 50th, 75th, 85th, 90th, and 95th percentiles.

The resulting fitted values, together with the original raw data, were then plotted against age to create the percentile curves separately constructed by race African American and Caucasian and gender. BMI percentiles were constructed for Caucasian children ages 5—15 years. The curves for African Americans and for the remainder of the obesity measures were limited to 5—12 year old children due to the sparsity of the data outside this age range.

Due to lack of adequate sample size, the 90th percentile of waist circumference for Caucasian girls, and the 95th percentile for the African American girls could not be accurately calculated. The percentile curves for a Caucasian boys, b Caucasian girls, c African American boys, and d African American girls are illustrated in Online Repository Figures 1—4 BMI percentiles, waist circumference, waist to height ratio, and conicity index respectively.

In addition, the 25th, 75th, 85th, and 95th percentile distributions of each obesity measure are presented by age and gender in Caucasians Table Ia online repository and African Americans Table Ib , online repository.

All the above analyses were conducted using SAS version 9. Structural Equation Modeling In order to account for the mediating effects of the obesity measures in the context of asthma and allergic sensitization, a structural equation model was developed using the Mplus software Further detail of the structural equation modeling is available in the on-line repository. Of the children with allergic rhinitis, No statistical differences in gender, parental education, household member smoking, or annual household income were found between the cases and controls.

In fact, i. Moreover, these mice showed no physical signs of anaphylaxis Fig. After sensitization, similar Fel d1—specific IgG titers were measured in all animals. Sensitized and nonvaccinated mice challenged with PBS or Fel d1 were used as controls. S4 and S5. As shown in Fig.

Similar results were obtained with purified total IgG unpublished data. Thus, Fel d1—specific IgGs are key effector molecules in the desensitization process and are able to block IgE-mediated mast cell degranulation.

In line with previous studies Takai et al. As reported previously Takai et al. These VLPs elicit strong B cell responses as a result of their highly organized and repetitive structures Jegerlehner et al. This feature can be exploited to enhance the immunogenicity of self- and foreign antigens. Chemical coupling of antigens via a cysteine to surface lysine residues on the VLP renders these antigens equally repetitive and consequently immunogenic.

Specifically, although a strong boosting of the IgE response was observed in sensitized control animals, no increase in sensitized and vaccinated mice was observed upon subsequent allergen challenge. In this case, inhibition would specifically affect IgE and not IgG B cell responses because antigen-specific IgG is strongly boosted upon vaccination.

This suggests that the reduced Th2 response is not caused by a shift to Th1 cells. The mechanism of this inhibition will need to be further investigated. It is possible that only moderate, and not strong, cross-linking of surface Ig is able to drive IgE responses. Indeed, viruses rather than parasites exhibit highly organized surface structures in a manner similar to Fel d1 displayed on VLPs.

IgE responses, however, protect against parasitic rather than viral infections. Thus, the combination of high cross-linking activity with TLR7 ligands may be able to block IgE memory responses long term. Indeed, some of our preliminary results indicated that the shutdown of the allergen-specific IgE response was the result of the single-stranded RNA within the VLPs. Shutting down this allergen-induced IgE response may therefore facilitate long-term curing of the patients. Indeed, seasonal allergen exposure usually boosts IgE responses in allergic individuals, maintaining or even worsening the disease.

The observation that repetitive display of Fel d1 on the VLPs resulted in reduced ability to trigger mast cell degranulation may seem somewhat unexpected. Hence, allergens displayed on VLPs are nonphysiological ligands for mast cells and may, therefore, not be able to properly activate the degranulation cascade. Furthermore, because of the linkage of the allergen to the relatively large VLPs 30 nm , the in vivo distribution will be fundamentally different from free Fel d1.

In this context, T reg cells have been recently described as a regulator of mast cell activity by altering IgE receptor expression and signaling Kashyap et al. T reg cells may also activate mast cells to mediate local immune suppression Lu et al. In addition, the regulatory cytokine IL, which is produced by T reg cells, has been shown to silence mast cell function in vivo and in vitro Grimbaldeston et al.

In contrast to these studies, we have found that our VLP-based vaccination primarily works through the induction of allergen-specific IgGs rather than T reg cells. Although successful immunotherapy in humans may involve both T and B cells Till et al. Antigen-specific IgG1 was shown to induce anaphylaxis in several in vitro and in vivo studies Ovary et al. Yet we find in this paper that polyclonal antibodies specific for Fel d1 dominated by the IgG1 isotype not only fail to sensitize mice for anaphylaxis but even prevent IgE-mediated anaphylaxis upon allergen challenge.

This difference may have several explanations. In the studies where IgG1-mediated anaphylaxis was observed, the reaction was induced by passive transfer of either antigen-specific IgE or IgG. In contrast to these studies, we used active sensitization with Fel d1 in WT mice. In addition, vaccination of sensitized mice results in specific IgG1 as well as IgG2a antibodies.

The presence of Fel d1—specific IgG2a antibodies may have masked a potential ability of IgG1 antibodies to trigger anaphylactic reactions. Interestingly, we found that IgG antibodies may inhibit the allergic reaction dependent on the allergen localization.

This suggests that because of the quick diffusion upon systemic exposure complete neutralization of the allergen may not be efficient by the circulating antibodies and some Fel d1 molecules will reach basophils. This suggests that Fel d1, which slowly diffuses within tissues, is recognized by specific IgG antibodies, covering up all available epitopes on the allergen, and may even result in local immune complexes, sequestering the allergen.

Hence, Fel d1—specific IgG antibodies may simply neutralize the allergen in the periphery. It has also been shown that serum obtained from subjects after antigen-specific immunotherapy could inhibit IgE-facilitated presentation of allergen to grass pollen—specific van Neerven et al.

In summary, our data demonstrate that allergens displayed on VLPs have the potential to rapidly treat established allergies and delineate two different mechanisms how IgG antibodies are able to block type I allergic reactions. All animals were used for experimentation according to protocols approved by the Swiss Federal Veterinary Office. The reaction was performed for 24 h immediately after elution of rFel d1 in the elution buffer by adding 2.

After covalent coupling, noncoupled rFel d1 was removed by gel filtration. After a min incubation at room temperature, cells were washed twice with PBS and analyzed by flow cytometry. Serial dilutions of sera were added to the plates and incubated for 2 h at room temperature. Optical densities were measured at nm. After a final wash with PBST, OPD substrate was added and incubated for at least 30 min at room temperature before optical densities were measured at nm.

Half-maximal antibody titers are defined as the reciprocal of the dilution leading to half of the OD measured at saturation. All standard dilutions were prepared in pyrogen-free glass tubes. The kinetic measurement with s intervals was performed at OD nm. For Fel d1 sensitization, 6-wk-old naive mice were injected i. For efficacy experiments, sensitized mice were vaccinated once s. Mice were injected i. Pricks through the ear skin were performed with 23G 0. For intradermal ear prick tests, an rFel d1 solution 0.

Dye extravasations started immediately after antigen challenge. To this end, the area of dye leakage was defined within the densitometer by gating around the leaked dye. A combination of size and color intensity of the pricked area defines the strength of dye leakage per ear, which is expressed as net intensity by the densitometer. Mean net intensity of different mice per group is shown. Acute systemic anaphylaxis.

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