Bystander effect in synergy to anergy in oral tolerance of Blomia tropicalis/ovalbumin murine co-immunization model.

Oral tolerance is an important approach in allergic diseases and murine model can provide useful information to improve its understanding and therapeutic measures. To address the influence of non-related allergen sensitization in immunized mice with the mite Blomia tropicalis (Bt) or ovalbumin (OVA) or with both Bt/OVA allergens. Furthermore, we sought to verify oral tolerance effect in the Bt/OVA co-immunization model. Mice sensitized with Bt and then exposed to OVA developed an enhanced IgE response to both allergens; contrariwise, this effect was not observed when OVA-sensitization was prior to Bt-sensitization. Co-injection of Bt and OVA led to a dominant IgE response towards OVA over Bt, which was not observed when co-immunization was performed with a 240-fold less amount of OVA. Induction of oral tolerance with OVA, prior to co-immunization, suppressed IgE response to both allergens, probably as a consequence of the increased levels of IFN-gamma found in these animals. The results evidenced that, depending on allergenic potential, new allergen exposure may exert an adjuvant effect to the first allergen used in the sensitization. The bystander suppression to non-related allergens through oral tolerance should be a useful mechanism to control sensitization to new allergens.

Blomia tropicalis and Dermatophagoides pteronyssinus mites evoke distinct patterns of airway cellular influx in type I hypersensitivity murine model.

Murine models of hypersensitivity to allergens are useful tools for the evaluation of strategies to downmodulate IgE response. We sought to compare allergen inflammatory pulmonary response in previously sensitized mice orally administered with dust mites Blomia tropicalis (Bt) or Dermatophagoides pteronyssinus (Dp). Sensitized A/Sn mice fed with Bt or Dp showed a significant decrease in the IgE response compared to control-immunized mice. Bt-immunized mice demonstrated an accumulation of neutrophils in the bronchoalveolar lavage fluid, while Dp-immunized mice revealed an intense influx of eosinophils in the airway. Bt oral administration did not attenuate cell influx in the airway and Dp-fed mice showed a significant decrease of neutrophils and lymphocytes in the bronchoalveolar lavage fluid. These findings demonstrated that oral tolerance induction to Bt and Dp extract in sensitized mice decrease IgE response, but does not interfere in local inflammatory pulmonary response. The distinct profile of airway cellular infiltration between mites immunization suggest an interesting model to study allergic inflammation.

IgA response in serum and gut secretion in sensitized mice fed with the dust mite Dermatophagoides pteronyssinus extract.

Induced oral tolerance to mucosal-exposed antigens in immunized animals is of particular interest for the development of immunotherapeutic approaches to human allergic diseases. This is a unique feature of mucosal surfaces which represent the main contact interface with the external environment. However, the influence of oral tolerance on specific and natural polyreactive IgA antibodies, the major defense mechanism of the mucosa, is unknown. We have shown that oral administration of an extract of the dust mite Dermatophagoides pteronyssinus (Dp) to primed mice caused down-regulation of IgE responses and an increase in tumor growth factor-beta secretion. In the present study, we observed that primed inbred female A/Sn mice (8 to 10 weeks old) fed by gavage a total weight of 1.0-mg Dp extract on the 6th, 7th and 8th days post-immunization presented normal secretion of IL-4 and IL-10 in gut-associated lymphoid tissue and a decreased production of interferon gamma induced by Dp in the draining lymph nodes (13,340 +/- 3,519 vs 29,280 +/- 2,971 pg/ml). Mice fed the Dp extract also showed higher levels of serum anti-Dp IgA antibodies and an increase of IgA-secreting cells in mesenteric lymph nodes (N = 10), reflecting an increase in total fecal IgA antibodies (N = 10). The levels of secretory anti-Dp IgA antibodies increased after re-immunization regardless of Dp extract feeding. Oral tolerance did not interfere with serum or secretory IgA antibody reactivity related to self and non-self antigens. These results suggest that induction of oral tolerance to a Dp extract in sensitized mice triggered different regulatory mechanisms which inhibited the IgE response and stimulated systemic and secretory IgA responses, preserving the natural polyreactive IgA antibody production.

IgA response in serum and gut secretion in sensitized mice fed with the dust mite Dermatophagoides pteronyssinus extract

Induced oral tolerance to mucosal-exposed antigens in immunized animals is of particular interest for the development of immunotherapeutic approaches to human allergic diseases. This is a unique feature of mucosal surfaces which represent the main contact interface with the external environment. However, the influence of oral tolerance on specific and natural polyreactive IgA antibodies, the major defense mechanism of the mucosa, is unknown. We have shown that oral administration of an extract of the dust mite Dermatophagoides pteronyssinus (Dp) to primed mice caused down-regulation of IgE responses and an increase in tumor growth factor-á secretion. In the present study, we observed that primed inbred female A/Sn mice (8 to 10 weeks old) fed by gavage a total weight of 1.0-mg Dp extract on the 6th, 7th and 8th days post-immunization presented normal secretion of IL-4 and IL-10 in gut-associated lymphoid tissue and a decreased production of interferon gamma induced by Dp in the draining lymph nodes (13,340 ñ 3,519 vs 29,280 ñ 2,971 pg/ml). Mice fed the Dp extract also showed higher levels of serum anti-Dp IgA antibodies and an increase of IgA-secreting cells in mesenteric lymph nodes (N = 10), reflecting an increase in total fecal IgA antibodies (N = 10). The levels of secretory anti-Dp IgA antibodies increased after re-immunization regardless of Dp extract feeding. Oral tolerance did not interfere with serum or secretory IgA antibody reactivity related to self and non-self antigens. These results suggest that induction of oral tolerance to a Dp extract in sensitized mice triggered different regulatory mechanisms which inhibited the IgE response and stimulated systemic and secretory IgA responses, preserving the natural polyreactive IgA antibody production.

Influence of maternal murine immunization with Dermatophagoides pteronyssinus extract on the type I hypersensitivity response in offspring.

BACKGROUND: Maternal exposure to environmental ubiquitous allergens could exert an influence on the newborn's immune repertoire and the later development of allergy. The aim of this study was to investigate the effects of maternal immunization with Dermatophagoides pteronyssinus (Dp) on the hypersensitivity response and IgG subclass production in offspring using a murine model. METHODS: A/Sn mice were immunized with Dp before mating with normal A/Sn males. Diaplacental serum samples were collected from newborn mice delivered by cesarean section, and maternal milk samples were extracted from the stomachs of newborn mice. Groups of offspring 25 or 45 days old were Dp immunized and boosted on the 10th day after sensitization. The animals were bled 7 days after the booster. RESULTS: High levels of anti-Dp IgG subclasses - mainly IgG1, but also IgG2a and IgG2b - were transmitted by immunized mice via the placenta to the offspring. In the milk from immunized mothers, significant levels of anti-Dp IgG subclasses and anti-Dp IgM and IgA antibodies were detected. Moreover, the increase in total IgA antibodies in the milk of the immunized females correlated with a significantly increased level of TGF-beta1. TGF-beta2 levels were markedly higher than the beta1 isoform in the milk, although no difference was observed between the groups. When offspring from immunized mothers were sensitized at 25 days, a significant decrease in total and anti-Dp IgE antibodies as well as total and anti-Dp IgG1, IgG2a and IgG2b subclasses was observed compared to normal female offspring, whereas when offspring were sensitized at 45 days, both offspring groups showed similar levels of IgE and IgG subclasses. CONCLUSIONS: Our study showed that maternal immunization with Dp promotes the transference of specific antibodies and/or TGF-beta, which can negatively modulate the allergic response in offspring, and suggests that maternal preexposure to allergen before mating can protect mice during the early phase.

Oral tolerance induction to Dermatophagoides pteronyssinus and Blomia tropicalis in sensitized mice: occurrence of natural autoantibodies to immunoglobulin E.

BACKGROUND: The dust mites Dermatophagoides pteronyssinus (Dp) and Blomia tropicalis (Bt) are important sources of indoor allergens in tropical and subtropical countries. Murine models allow the analysis of the immune response and regulation of IgE production to Dp and Bt allergens. Oral tolerance induces unresponsiveness in naive animals, but its application in sensitized animals can provide useful information to improve allergy therapy. OBJECTIVE: To study the profile of IgE and IgG subclasses antibody upon oral administration with Bt and Dp extract in previously sensitized mice. Further, the occurrence of autoantibodies IgG anti-IgE in the immunization and in the oral tolerance was investigated. METHODS: A/Sn mice were immunized with Bt or Dp extract in alum, orally administrated with 0.25 mg of Bt or Dp extract or PBS at the 6th, 7th and 8th days after immunization and boosted twice with their respective allergens. To analyse the mice groups, specific IgE antibodies were measured by passive anaphylaxis reaction and specific IgG subclasses and anti-IgE IgG autoantibody by ELISA assay. RESULTS: IgE levels were markedly increased in Bt-immunized mice compared with Dp-immunized mice. A distinct profile of the specific isotypes was verified in Bt-immunized mice with a preferential production of IgG3 and IgA antibodies, whereas Dp-immunized mice developed high titres of anti-Dp IgG1, IgG2a and IgG2b antibodies. The antigen feeding inhibited IgE response in both fed-mice groups but only Dp-fed mice presented decreased levels of IgG antibodies. Free anti-IgE IgG autoantibodies were detected mainly in the Dp-immunization and they correlated with the antibody isotypes found against the allergen. CONCLUSIONS: This is the first time that the murine-type I hypersensitivity is employed to study Bt-immunization, showing a marked IgE production, associated with IgG response, which is at least in part driven by T-independent antigens. The oral tolerance protocol in previously sensitized animals was able to down-modulate IgE response and points out this route as a strategy for allergy therapy.

Oral tolerance induction in dermatophagoides pteronyssinus-sensitized mice induces inhibition of IgE response and upregulation of TGF-beta secretion.

Oral antigen administration induces peripheral tolerance in naive animals. Studies of oral tolerance induction in sensitized mice have clinical relevance as a strategy to modulate allergy. In this study, the A/Sn mice sensitized with extract of Dermatophagoides pteronyssinus (Dp) and submitted to oral Dp administration showed a marked decrease in IgE anti-Dp antibody production compared with sensitized phosphate-buffered saline (PBS)-fed mice. T cells from Dp-fed mice cocultured with spleen cells from PBS-fed mice were able to inhibit IgE anti-Dp antibody production and did not interfere in IgG1 antibody levels. The analysis of cytokine profile after Dp feeding showed a significant decrease in interleukin-4 (IL-4), IL-5, and IL-13 antigen-induced secretion levels by spleen cells, without shifting to IL-2 and interferon-gamma (IFN-gamma) production. Both transforming growth factor-beta (TGF-beta) baseline and TGF-beta antigen-stimulated levels were increased in Dp-fed mice. The effects of regulatory cytokines on anti-Dp IgE antibody production were investigated in vitro. The addition of recombinant TGF-beta (rTGF-beta) to spleen cell cultures stimulated by Dp inhibited IgE antibody secretion in both mouse groups. Neutralizing antibodies to IL-4, but not anti-TGF-beta, induced a marked inhibition of IgE production. Therefore, a negative modulatory effect on IgE response by inhibition of the axis Th2 was observed in sensitized Dp-fed mice, possibly mediated by induction of regulatory cytokines.

Modulation of IgE response and cytokine production in Peyer's patches and draining lymph nodes in sensitized mice made tolerant by oral dust mite administration.

Such allergic diseases as rhinitis and asthma are IgE-mediated type I reactions and are controlled primarily by Th2 cells. One of the major dust mites, Dermatophagoides pteronyssinus (Dp), is considered to cause allergic reactions. Oral tolerance, largely used to modulate immune response, opens the possibility of modulating Th2 allergic responses. We observed downmodulation of total and specific IgE antibody levels as well as the number of specific IgE-secreting cells with Dp feeding in previously sensitized mice. Analysis of the cytokine profile in mucosal lymphoid tissues in the protocol revealed altered patterns of interferon-gamma (IFN-gamma), interleukin-5 (IL-5), and transforming growth factor-beta (TGF-beta) secretion in Dp-fed animals. The results suggest that both the Th and B cell populations are modulated in mice made tolerant by oral Dp feeding. Understanding the mechanisms at the mucosal level that underlie oral tolerance can improve its use in allergy immunotherapy.

Low dose of orally administered antigen down-regulates the T helper type 2-response in a murine model of dust mite hypersensitivity.

One of the main goals of immunotherapy of allergic diseases is the down-regulation of the type I hypersensitivity reaction. We investigated in this study the effect of oral administration of varying doses (0.25, 1.0, 4.0 and 10 mg) of dust mite extract (Dermatophagoides pteronyssinus, Dp) in sensitized A/Sn mice. A marked decrease of the allergen-specific immunoglobulin E (IgE) response was observed with all antigen doses. The mice orally tolerized with low Dp dose (0.25 mg) had a significant decrease in the total serum IgE and in the immunoglobulin G1 (IgG1), IgG2a and IgG2b antibody levels. The higher Dp dose (10.0 mg), however, enhanced the IgG1 antibody response, suggesting the stimulation of a pre-existing immune response of the sensitized animals. Animals fed with the low Dp dose had a significant decrease in the frequency of interleukin-4 (IL-4) secreting cells. These animals also showed a significant decrease in the frequency of Dp-specific IgE- and IgG1-positive plasma cells. Our data suggest that feeding dust mite extract to Dp-sensitized mice down-regulates the development of type I hypersensitivity, by inhibition of the T helper 2 response.

Oral tolerance induced to house dust mite extract in naive and sensitized mice: evaluation of immunoglobulin G anti-immunoglobulin E autoantibodies and IgG-IgE complexes.

We investigated the effect on specific antibody response of naive and sensitized mice orally administrated with low (0.25 mg) or high (10.0 mg) doses of Dermatophagoides pteronyssinus (Dp) extract. We also examined the effect of oral administration of Dp on the production of autoantibodies to immunoglobulin G (IgG) and immunoglobulin E (IgE). Naive and sensitized mice both showed a marked down-regulation of IgE antibody production, regardless of the dose of Dp. We also detected an inhibitory effect of the total IgE levels and the allergen-specific IgG1, IgG2a and IgG2b antibody response in sensitized mice given the low dose of Dp. In contrast, high doses of Dp stimulated IgG1 antibody production in both naive and sensitized animals. In addition, the oral tolerance induction protocol stimulated anti-F(ab')2gamma and anti-Fcgamma autoantibody production. Evaluation of IgG anti-IgE autoantibodies by a direct enzyme immunoassay (EIA) revealed the presence of these autoantibodies, predominantly of the IgG1 isotype, specifically in those animals fed with the high dose. In contrast, IgG-IgE complexes, determined by EIA using immobilized anti-IgE antibodies, were detected mainly in sera of control animals. The autoantibody anti-IgE specificity was tested against IgE-TNP and IgE-DANSYL murine proteins and revealed different inhibition profiles, suggesting the action of heterogeneous subpopulations of autoantibodies. Taken together, our results show that the oral tolerance protocol with Dp was able to modulate the production of allergen-specific IgE antibodies in both naive and sensitized animals. In addition, we suggest that anti-IgE autoantibodies participate in the modulation of allergic response triggered by oral tolerance protocols.