Co-administration Of Ovalbumin Research Articles

Nicotine, as a novel tolerogenic adjuvant, enhances the efficacy of immunotherapy in a mouse model of allergic asthma

An increasing trend in the incidence of allergic diseases including asthma and related morbidity and mortality is observed worldwide during the last decades. Allergen-specific immunotherapy is suggested for the treatment of some allergic diseases; nevertheless, there is always a menace of uncommon, but life-treating reactions due to increasing the administration of allergen extract doses. Hence, improving its efficacy may reduce the required doses as well as the risk of such reactions. The current study aimed at examining the effects of nicotine (NIC), as a tolerogenic adjuvant, on the improvement of immunotherapy efficacy in a mouse model of allergic asthma. BALB/c mice were sensitized using alum and ovalbumin (OVA) on the days 0 and 7. Mice received OVA either alone or together with NIC (1 or 10 mg/kg) on the days 21, 23, and 25. Then, the mice were challenged with OVA 5% using a nebulizer on the days 35, 38, and 41 and sacrificed the next day. Co-administration of OVA and NIC decreased the inflammation of the lung tissue, eosinophils count in the bronchoalveolar lavage (BAL) fluid, the serum level of OVA-specific immunoglobulin E, as well as interleukin (IL)-4 production, while increasing the population of antigen-specific regulatory T-cells (Treg cells) and transforming growth factor-β/IL-4 (TGF-β/IL-4) ratio compared to the OVA and control groups in a dose-dependent manner. Collectively, the findings suggest that administration of NIC plus the allergen increased immunotherapy efficacy through decreasing allergic inflammation and allergic responses intensity, and increasing Treg cells population.

Intestinal Epithelial Cells Regulate Gut Eotaxin Responses and Severity of Allergy.

Intestinal epithelial cells (IECs) are known to regulate allergic sensitization. We addressed the role of the intrinsic IKKβ signaling in IECs in the effector phase of allergy following oral allergen challenge and its impact on the severity of responses is poorly. Upon orally sensitization by co-administration of ovalbumin with cholera toxin as adjuvant, wild-type and mice lacking IKKβ in IECs (IKKβΔIEC mice) developed similar levels of serum IgE and allergen-specific secretory IgA in the gut. However, subsequent allergen challenges in the gut promoted allergic lower responses in KKβΔIEC mice. Analysis of cytokines and chemokines in serum and gut tissues after oral allergen challenge revealed impaired eotaxin responses in IKKβΔIEC mice, which correlated with lower frequencies of eosinophils in the gut lamina propria. We also determined that IECs were a major source of eotaxin and that impaired eotaxin production was due to the lack of IKKβ signaling in IECs. Oral administration of CCL11 to IKKβΔIEC mice during oral allergen challenge enhanced allergic responses to levels in wild-type mice, confirming the role of IEC-derived eotaxin as regulator of the effector phase of allergy following allergen challenge. Our results identified targeting IEC-derived eotaxin as potential strategy to limit the severity of allergic responses to food antigens.

Coadministration of a Plasmid Encoding HIV-1 Gag Enhances the Efficacy of Cancer DNA Vaccines.

DNA vaccination holds great promise for the prevention and treatment of cancer and infectious diseases. However, the clinical ability of DNA vaccines is still controversial due to the limited immune response initially observed in humans. We hypothesized that electroporation of a plasmid encoding the HIV-1 Gag viral capsid protein would enhance cancer DNA vaccine potency. DNA electroporation used to deliver plasmids in vivo, induced type I interferons, thereby supporting the activation of innate immunity. The coadministration of ovalbumin (OVA) and HIV-1 Gag encoding plasmids modulated the adaptive immune response. This strategy favored antigen-specific Th1 immunity, delayed B16F10-OVA tumor growth and improved mouse survival in both prophylactic and therapeutic vaccination approaches. Similarly, a prophylactic DNA immunization against the melanoma-associated antigen gp100 was enhanced by the codelivery of the HIV-1 Gag plasmid. The adjuvant effect was not driven by the formation of HIV-1 Gag virus-like particles. This work highlights the ability of both electroporation and the HIV-1 Gag plasmid to stimulate innate immunity for enhancing cancer DNA vaccine immunogenicity and demonstrates interesting tracks for the design of new translational genetic adjuvants to overcome the current limitations of DNA vaccines in humans.

Epicutaneous Allergic Sensitization by Cooperation between Allergen Protease Activity and Mechanical Skin Barrier Damage in Mice

Allergen sources such as mites, insects, fungi, and pollen contain proteases. Airway exposure to proteases induces allergic airway inflammation and IgE/IgG1 responses via IL-33-dependent mechanisms in mice. We examined the epicutaneous sensitization of mice to a model protease allergen, papain; the effects of tape stripping, which induces epidermal barrier dysfunction; and the atopic march upon a subsequent airway challenge. Papain painting on ear skin and tape stripping cooperatively promoted dermatitis, the skin gene expression of proinflammatory cytokines and growth factors, up-regulation of serum total IgE, and papain-specific IgE/IgG1 induction. Epicutaneous sensitization induced T helper (Th) 2 cells and Th17 differentiation in draining lymph nodes. Ovalbumin and protease inhibitor-treated papain induced no or weak responses, whereas the co-administration of ovalbumin and papain promoted ovalbumin-specific IgE/IgG1 induction. Wild-type and IL-33-deficient mice showed similar responses in the epicutaneous sensitization phase. The subsequent airway papain challenge induced airway eosinophilia and maintained high papain-specific IgE levels in an IL-33-dependent manner. These results suggest that allergen source-derived protease activity and mechanical barrier damage such as that caused by scratching cooperatively promote epicutaneous sensitization and skin inflammation and that IL-33 is dispensable for epicutaneous sensitization but is crucial in the atopic march upon a subsequent airway low-dose encounter with protease allergens.

Glyphosate-rich air samples induce IL-33, TSLP and generate IL-13 dependent airway inflammation.

Several low weight molecules have often been implicated in the induction of occupational asthma. Glyphosate, a small molecule herbicide, is widely used in the world. There is a controversy regarding a role of glyphosate in developing asthma and rhinitis among farmers, the mechanism of which is unexplored. The aim of this study was to explore the mechanisms of glyphosate induced pulmonary pathology by utilizing murine models and real environmental samples. C57BL/6, TLR4-/-, and IL-13-/- mice inhaled extracts of glyphosate-rich air samples collected on farms during spraying of herbicides or inhaled different doses of glyphosate and ovalbumin. The cellular response, humoral response, and lung function of exposed mice were evaluated. Exposure to glyphosate-rich air samples as well as glyphosate alone to the lungs increased: eosinophil and neutrophil counts, mast cell degranulation, and production of IL-33, TSLP, IL-13, and IL-5. In contrast, in vivo systemic IL-4 production was not increased. Co-administration of ovalbumin with glyphosate did not substantially change the inflammatory immune response. However, IL-13-deficiency resulted in diminished inflammatory response but did not have a significant effect on airway resistance upon methacholine challenge after 7 or 21 days of glyphosate exposure. Glyphosate-rich farm air samples as well as glyphosate alone were found to induce pulmonary IL-13-dependent inflammation and promote Th2 type cytokines, but not IL-4 for glyphosate alone. This study, for the first time, provides evidence for the mechanism of glyphosate-induced occupational lung disease.

Basophils are recruited to inflamed lungs and exacerbate memory Th2 responses in mice and humans

Although the contribution of basophils as inducers or amplifiers of Th2 responses is still debated, prolonged basophil/CD4 T cell interactions were observed in lungs but not lymph nodes (LNs) of parasite-infected mice. However, the impact of basophils on the function of tissue CD4 effector T cells remains unknown. Basophils were purified from the lungs of ovalbumin (OVA)-sensitized and OVA-challenged (OVA-immunized) mice or human peripheral blood for in vivo and in vitro functional studies. Pulmonary basophils were adoptively transferred to OVA-sensitized hosts to assess airway inflammation in bronchoalveolar lavage fluid (BALF) and Th2 responses in lung explants and draining LNs. Basophils were co-cultured with effector T cells or Ag-specific naïve T cells alone or in combination with dendritic cells (DCs); IL-4 production was determined by flow cytometry and ELISA. Basophils accumulated in lungs of OVA-immunized mice. Adoptive transfer of basophils to OVA-sensitized hosts enhanced lung IL-4 and IL-13 release while co-administration of OVA further aggravated airway inflammation and Th2 responses in LNs. Mechanistic in vitro studies revealed that pulmonary basophils interacted with lung CD4 effectors, in the absence of DCs, to increase T cell survival and Th2 cytokine expression at the single cell level but amplified OVA-loaded DC-driven Th2 differentiation. Finally, human basophils augmented in vitro IL-4 expression in effector memory CD4 T cells that include CRTH2(+) cells through IL-4 and TCR-independent pathways. Basophils may worsen Th2 inflammatory disorders through direct interactions with pathogenic CD4 T cells as well as by enhancing DC-induced Th2 cell development.

Enhancement of Gastrointestinal Absorption of Ovalbumin Caused by Spermine Induces an Increase in Plasma Histamine Levels in Mice Sensitized to Ovalbumin

The aim of this study was to determine whether a spermine (SPM)-induced increase in gastrointestinal absorption of an allergen leads to an anaphylactic response in sensitized mice. First, we examined the enhancing effect of SPM on the gastrointestinal absorption of ovalbumin (OVA) in an in situ jejunum loop study in rats and an in vivo oral absorption study in mice. Second, we investigated whether enhancement of gastrointestinal absorption of OVA caused by SPM induces an anaphylactic response in mice sensitized to OVA. In the in situ jejunum loop study in rats, a significant amount of immune-reactive OVA was detected in the plasma after co-administration of OVA and SPM. Oral co-administration of OVA and SPM to mice in vivo also increased plasma OVA concentrations in an SPM dose-dependent manner. Furthermore, in sensitized mice, a significant increase in plasma histamine levels occurred along with the increase in plasma OVA levels after co-administration of OVA with SPM. This finding suggests that an SPM-induced increase in gastrointestinal absorption of OVA leads to an anaphylactic response. These results indicate that excess oral ingestion of SPM may have widespread health effects, including the induction of food allergies, via modulation of the function of the gastrointestinal epithelial barrier.

Paclitaxel acts as an adjuvant to promote both Th1 and Th2 immune responses induced by ovalbumin in mice

Paclitaxel, a diterpenoid isolated from the bark of the Taxus cuspidate cv. Nana, was evaluated for its adjuvant effect on the immune responses in a mouse model. Fifty-six mice were randomly distributed into seven groups with 8 mice in each. Animals were subcutaneously immunized on days 1 and 21 with 100 μg of paclitaxel, 10 μg of ovalbumin (OVA), OVA with paclitaxel (50, 100 or 200 μg) or with aluminum hydroxide (alum). Two weeks after the primary and boost immunizations, blood samples were collected for measurement of serum antibodies. Splenocytes were separated for detection of lymphocyte proliferation in responses to concanavalin A (Con A), lipopolysaccharide (LPS) and OVA, and mRNA expression of Th1 cytokines (IFN-γ and IL-12), Th2 cytokines (IL-10 and IL-5) and transcription factors T-bet/GATA-3 (Th1/Th2 switcher). Results showed that coadministration of OVA with paclitaxel induced significantly higher IgG, IgG1, IgG2a, IgG2b, IgG3 and IgM responses than when OVA was used alone. In addition, up-regulated T-bet/GATA-3 together with significantly increased mRNA expression of IL-4, IL-10, IFN-γ and IL-12 by splenocytes, as well as the proliferative responses of splenocytes to Con A, LPS and OVA were observed in paclitaxel-adjuvanted groups. Incubation of a murine macrophage-like cell line with paclitaxel significantly increased TNF-α and -10 released from the cells and expression of microRNAs such as miR-155, miR-147, miR-146a and miR-132. Therefore, paclitaxel activated both Th1 and Th2 responses. Considering its unique adjuvant effect demonstrated in this study and a safe record clinically used as an antineoplastic agent, paclitaxel could be an ideal adjuvant candidate when mixed Th1/Th2 immune responses are needed.

Cupressaceae Pollen Grains Modulate Dendritic Cell Response and Exhibit IgE-Inducing Adjuvant Activity In Vivo

Pollen is considered a source of not only allergens but also immunomodulatory substances, which could play crucial roles in sensitization and/or the exacerbation of allergies. We investigated how allergenic pollens from different plant species (Japanese cedar and Japanese cypress, which belong to the Cupressaceae family, and birch, ragweed, and grass) modulate murine bone marrow-derived dendritic cell (DC) responses and examined the effect of Cupressaceae pollen in vivo using mice. DCs were stimulated with pollen extracts or grains in the presence or absence of LPS. Cell maturation and cytokine production in DCs were analyzed by flow cytometry, ELISA, and/or quantitative PCR. Pollen extracts suppressed LPS-induced IL-12 production and the effect was greatest for birch and grass. Without LPS, pollen grains induced DC maturation and cytokine production without IL-12 secretion and the response, for which TLR 4 was dispensable, was greatest for the Cupressaceae family. Intranasal administration of Cupressaceae pollen in mice induced an elevation of serum IgE levels and airway eosinophil infiltration. Coadministration of ovalbumin with Cupressaceae pollen grains induced ovalbumin-specific IgE responses associated with eosinophil infiltration. The results suggest that modulation of DC responses by pollen differs among the plant families via (1) the promotion of DC maturation and cytokine production by direct contact and/or (2) the inhibition of IL-12 production by soluble factors. The strong DC stimulatory activity in vitro and IgE-inducing activity in mice support the clinical relevance of Cupressaceae pollen to allergies in humans.

T helper 1 cells stimulated with ovalbumin and IL-18 induce airway hyperresponsiveness and lung fibrosis by IFN-γ and IL-13 production

We previously reported that ovalbumin (OVA) and IL-18 nasally administered act on memory type T helper (Th)1 cells to induce airway hyperresponsiveness (AHR) and inflammation, which is characterized by peribronchial infiltration with neutrophils and eosinophils. Here, we report this administration also induces lung fibrosis in an IL-13-dependent manner. Th1 cells secrete several cytokines, including IFN-gamma and bronchogenic cytokine IL-13, when stimulated with antigen (Ag) and IL-18. However, IL-13 blockade failed to attenuate AHR, although this treatment inhibited eosinophilic infiltration. To understand the mechanism by which Th1 cells induce AHR after Ag plus IL-18 challenge, we established "passive" and "active" Th1 mice by transferring OVA-specific Th1 cells into naïve BALB/c mice or by immunizing naïve BALB/c mice with OVA/complete Freund's adjuvant, respectively. Administration of Ag and IL-18 induced both types of Th1 mice to develop AHR, airway inflammation, and lung fibrosis. Furthermore, this treatment induced deposition of periostin, a novel component of lung fibrosis. Neutralization of IL-13 or IFN-gamma during Ag plus IL-18 challenges inhibited the combination of eosinophilic infiltration, lung fibrosis, and periostin deposition or the combination of neutrophilic infiltration and AHR, respectively. We also found that coadministration of OVA and LPS into Th1 mice induced AHR and airway inflammation via endogenous IL-18. Thus, IL-18 becomes a key target molecule for the development of a therapeutic regimen for the treatment of Th1-cell-induced bronchial asthma.

Inoculation Route-Dependent and Allergen-Specific Suppressive Effects of Bacille Calmette-Guérin Vaccination on Asthmatic Reactions in BALB/c Mice

Intranasal bacille Calmette-Guérin (BCG) vaccination causes greater suppression of ovalbumin-induced airway eosinophilia in mice than does subcutaneous vaccination. Coadministration of ovalbumin with interleukin (IL)-18 induces an ovalbumin-specific Th1 immune reaction. The purpose of this study was to examine whether the suppressive effect of BCG is dependent on the inoculation method, using various murine asthma models. Female BALB/c mice (n = 7 per group) were immunized with BCG subcutaneously or intranasally, then sensitized with ovalbumin or Dermatophagoides farinae either immediately or one week later. After provocation with one of the allergens, the mice were tested by methacholine bronchial challenge, and analyses of the inflammatory cell numbers in the airways and cytokine levels in the supernatant of concanavalin A-stimulated splenocytes were conducted. Overall, the airway responses to the allergens were significantly lower and the interferon (IFN)-gamma level was significantly higher in BCG-treated mice than in untreated mice, and the number of airway eosinophils was significantly related to the IFN-gamma/IL-5 ratio (r = -0.444, p < 0.001). Subcutaneous BCG inoculation tended to have a greater suppressive effect on the development of airway hyperresponsiveness and eosinophilia than did intranasal inoculation. Concurrent BCG vaccination and D. farinae sensitization one week before ovalbumin sensitization tended to have a greater suppressive effect on airway responsiveness to methacholine induced by D. farinae aerosols than did that induced by ovalbumin aerosols. Subcutaneous BCG inoculation suppressed asthmatic reactions more remarkably than did intranasal inoculation, and concurrent BCG vaccination and allergen sensitization induced allergen-specific suppression of asthmatic reactions.

Enhancement of ovalbumin-specific IgA responses via oral boosting with antigen co-administered with an aqueous Solanum torvum extract

An experiment was conducted with the objective to enhance mucosal immunity against ovalbumin (OVA) by co-administration of OVA with an aqueous extract from the fruit of Solanum torvum (STE). Five groups of female ICR mice aged approximately 8 weeks at the commencement of the experiment were caged in groups of eight and received various treatments. The treatments included OVA alone, OVA with cholera toxin (CT), and OVA with various doses of STE. Mice were primed intraperitoneally with 500 μg of OVA alone or co-administered with 0.1 μg CT, or with 1 μg STE. All mice were boosted orally via gastric intubation 14 days after priming with 10 mg OVA alone, or co-administered with 10 μg CT or with 10 mg, 1 mg or 0.1 mg STE. One week later all mice were killed and organs obtained for analysis of the immune response. Intestinal, faecal and pulmonary OVA-specific sIgA concentration was significantly increased (p<0.05) in mice that received booster combinations of OVA/CT and OVA with all extract doses (p<0.05). Specific serum IgG titres did not differ significantly between groups. It is concluded that STE can significantly enhance secretory immunity in the intestine to OVA with mucosal homing to the lungs. The adjuvant effect of STE is comparable to that of CT.

Enhancement of ovalbumin-specific IgA responses via oral boosting with antigen co-administered with an aqueous Solanum torvum extract

An experiment was conducted with the objective to enhance mucosal immunity against ovalbumin (OVA) by co-administration of OVA with an aqueous extract from the fruit of Solanum torvum (STE). Five groups of female ICR mice aged approximately 8 weeks at the commencement of the experiment were caged in groups of eight and received various treatments. The treatments included OVA alone, OVA with cholera toxin (CT), and OVA with various doses of STE. Mice were primed intraperitoneally with 500 μg of OVA alone or co-administered with 0.1 μg CT, or with 1 μg STE. All mice were boosted orally via gastric intubation 14 days after priming with 10 mg OVA alone, or co-administered with 10 μg CT or with 10 mg, 1 mg or 0.1 mg STE. One week later all mice were killed and organs obtained for analysis of the immune response. Intestinal, faecal and pulmonary OVA-specific sIgA concentration was significantly increased (p<0.05) in mice that received booster combinations of OVA/CT and OVA with all extract doses (p<0.05). Specific serum IgG titres did not differ significantly between groups. It is concluded that STE can significantly enhance secretory immunity in the intestine to OVA with mucosal homing to the lungs. The adjuvant effect of STE is comparable to that of CT.

Environmental aspects of skin diseases in children: management of common problems

Allergen sources such as mites, insects, fungi, and pollen contain proteases. Airway exposure to proteases induces allergic airway inflammation and IgE/IgG1 responses via IL-33–dependent mechanisms in mice. We examined the epicutaneous sensitization of mice to a model protease allergen, papain; the effects of tape stripping, which induces epidermal barrier dysfunction; and the atopic march upon a subsequent airway challenge. Papain painting on ear skin and tape stripping cooperatively promoted dermatitis, the skin gene expression of proinflammatory cytokines and growth factors, up-regulation of serum total IgE, and papain-specific IgE/IgG1 induction. Epicutaneous sensitization induced T helper (Th) 2 cells and Th17 differentiation in draining lymph nodes. Ovalbumin and protease inhibitor-treated papain induced no or weak responses, whereas the co-administration of ovalbumin and papain promoted ovalbumin-specific IgE/IgG1 induction. Wild-type and IL-33–deficient mice showed similar responses in the epicutaneous sensitization phase. The subsequent airway papain challenge induced airway eosinophilia and maintained high papain-specific IgE levels in an IL-33–dependent manner. These results suggest that allergen source-derived protease activity and mechanical barrier damage such as that caused by scratching cooperatively promote epicutaneous sensitization and skin inflammation and that IL-33 is dispensable for epicutaneous sensitization but is crucial in the atopic march upon a subsequent airway low-dose encounter with protease allergens.

Effect of organic acids, trypsin inhibitors and dietary protein on the pharmacological activity of recombinant human granulocyte colony-stimulating factor (rhG-CSF) in rats

The effects of organic acids, trypsin inhibitors and dietary protein on the oral availability of recombinant human granulocyte colony-stimulating factor (rhG-CSF) was investigated in rats by measuring the blood total leucocyte (BTL) counts. As organic acids, tartaric, succinic and citric acids were used at 2.30–7.68 mg/kg. Two trypsin inhibitors, soybean trypsin inhibitor (SBTI, 0.8–3.2 mg/kg) and chicken egg white trypsin inhibitor (CETI, 1.92 mg/kg), together with ovalbumin (0.80 mg/kg) as a representative of dietary proteins were employed with co-administration of citric acid (4.61 mg/kg). After intraduodenal administration of several kinds of test rhG-CSF solutions to rats, blood samples were collected for 48 h and the BTL counts were determined. The results are expressed as relative increase in BTL counts as compared to that before administration of the dose. The areas under the curves (AUC; % BTL increase × h) obtained for test solutions containing tartaric, succinic and citric acids were 2.1, 1.1 and 5.1 times greater than that of the control experiment, where rhG-CSF solution buffered with phosphate buffer (pH 6.1) was used. On administration of SBTI (3.20 mg/kg) and CETI (1.92 mg/kg) in combination with citric acid (4.61 mg/kg), the AUC values were determined to be 6.5 and 5.1 times greater than that of the control experiment, respectively. Co-administration of ovalbumin (0.80 mg/kg) with citric acid (4.61 mg/kg) also increased the AUC 6.3-fold as compared to the control. However, a low dose of SBTI (0.80 mg/kg) co-administered with citric acid did not exert a synergistic action.