ovalbumin and Dermatitis--Allergic-Contact

The pathological process of atopic dermatitis (AD) progressing into other types of allergic diseases such as asthma and allergic rhinitis during the first several years of life is often referred to as the atopic march. Although the phenomenon of atopic march has been recognized for decades, how asthma stems from AD is still not fully understood, confounding a universal strategy to effectively protect people from the atopic march.. We established experimental atopic march mice by first inducing allergic dermatitis with 0.5% fluorescein isothiocyante (FITC) applied to the skin, followed by an ovalbumin (OVA) airway challenge. In addition, by examining serum immunoglobulin (Ig) concentrations, airway cytokines, the levels of oxidative stress markers, histopathological changes in lung tissue and airway hyperresponsiveness (AHR), we were able to validate the successful establishment of the model. Furthermore, by detecting the attenuating effects of melatonin (MT) and the levels of oxidative stress in the atopic march mice, we explored the potential molecular mechanisms involved in the development of atopic march.. By successfully establishing an experimental atopic march mouse model, we were able to demonstrate that overproduction of oxidative stress in the lung significantly up-regulated the activation of nuclear factor-κB (NF-κB) signaling pathways causing thymic stromal lymphopoietin (TSLP) release, which further promotes the development of atopic march.. To mitigate the development of the atopic march, antioxidants such as MT may be imperative to inhibit NF-κB activation in the lung, especially after the onset of AD.

Topics: Animals; Anti-Inflammatory Agents; Cytokines; Dermatitis, Allergic Contact; Disease Models, Animal; Disease Progression; Fluorescein-5-isothiocyanate; Immunoglobulin E; Immunoglobulin G; Inflammation Mediators; Lung; Male; Melatonin; Mice, Inbred BALB C; NF-kappa B; Ovalbumin; Oxidative Stress; Pneumonia

Epithelial tissues continually undergo apoptosis. Commensal organisms that inhabit the epithelium influence tissue homeostasis, in which regulatory T cells (Treg cells) have a central role. However, the physiological importance of epithelial cell apoptosis and how the number of Treg cells is regulated are both incompletely understood. Here we found that apoptotic epithelial cells negatively regulated the commensal-stimulated proliferation of Treg cells. Gut commensals stimulated CX3CR1(+)CD103(-)CD11b(+) dendritic cells (DCs) to produce interferon-β (IFN-β), which augmented the proliferation of Treg cells in the intestine. Conversely, phosphatidylserine exposed on apoptotic epithelial cells suppressed IFN-β production by the DCs via inhibitory signaling mediated by the cell-surface glycoprotein CD300a and thus suppressed Treg cell proliferation. Our findings reveal a regulatory role for apoptotic epithelial cells in maintaining the number of Treg cell and tissue homeostasis.

Topics: Allergens; Animals; Apoptosis; Colitis; Colon; Dendritic Cells; Dermatitis, Allergic Contact; Dextran Sulfate; Epidermal Cells; Epidermis; Epithelial Cells; Flow Cytometry; Gastrointestinal Microbiome; Immunohistochemistry; Interferon-beta; Intestinal Mucosa; Langerhans Cells; Lung; Mice; Mice, Knockout; Ovalbumin; Real-Time Polymerase Chain Reaction; Receptors, Immunologic; Respiratory Mucosa; Salmonella Infections; Salmonella typhimurium; T-Lymphocytes, Regulatory

Topics: Allergens; Animals; Cytokines; Dermatitis, Allergic Contact; Eosinophils; Mice; Mice, Inbred BALB C; Mice, Knockout; Neutrophils; Ovalbumin; Receptors, Complement; RNA, Messenger; Vaccinia; Vaccinia virus; Viral Load

Annexin A1 (AnxA1) is recognized as an endogenous anti-inflammatory molecule. However, its effects on the adaptive immune response and, in particular, on T cells remain unclear. In this study, we investigated the actions of AnxA1 in three distinct models of T cell-mediated inflammation. In contact hypersensitivity, collagen-induced arthritis, and inflammation induced by OT-II TCR transgenic T cells responding to OVA, AnxA1 deficiency significantly increased Ag-induced T cell proliferation and the resultant level of inflammation. In the contact hypersensitivity model, this was associated with increased adhesion of CD4(+) T cells, CD8(+) T cells, and neutrophils in the dermal microvasculature, as well as increased T cell expression of RORγt and IL-17A. In collagen-induced arthritis, deficiency of endogenous AnxA1 increased susceptibility to arthritis and Ag-specific T cell activation. Deficiency of AnxA1 also increased OVA-induced cutaneous delayed-type hypersensitivity and IFN-γ and IL-17 release. Transfer experiments using CD4(+) T cells from AnxA1(-/-) mice demonstrated that the absence of AnxA1 solely in T cells resulted in increased inflammatory responses in wild-type recipients. Similarly, experiments using AnxA1(-/-) OT-II CD4(+) T cells demonstrated that the absence of AnxA1 in T cells was sufficient to induce increased Ag-specific CD4(+) T cell proliferation in vivo, augment T cell production of IFN-γ, IL-17, TNF, and IL-6, and increase Akt, ERK, and p38 activation. Together, these findings indicate that T cell-expressed AnxA1 functions to attenuate T cell-driven inflammatory responses via T cell-intrinsic effects on intracellular signaling, proliferation, and Th1/Th17 cytokine release.

Topics: Animals; Annexin A1; Arthritis, Experimental; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Adhesion; Dermatitis, Allergic Contact; Enzyme Activation; Gene Expression Regulation; Hypersensitivity, Delayed; Inflammation; Interleukin-17; Lymphocyte Activation; Lymphokines; Mice; Mice, Inbred C57BL; Mice, Knockout; Mice, Transgenic; Neutrophils; Nuclear Receptor Subfamily 1, Group F, Member 3; Ovalbumin; Oxazolone; Peptide Fragments; Signal Transduction; Specific Pathogen-Free Organisms

Allergic contact dermatitis (ACD) resulting from exposure to low molecular weight chemicals is a common clinical condition in industrialized countries and can be mediated by either Th1 or Tc1 lymphocytes. The animal model of contact sensitivity (CS) is commonly used to study ACD in mice and helps to test new therapeutics. We have previously shown that epicutaneous (EC) immunization with TNP-Ig prior to hapten sensitization inhibits Th1-mediated CS and observed that the suppression is mediated by TCRαβ(+) CD4(+) CD8(+) cells and is TGF-β dependent. More recently we have shown that EC immunization with DNP-BSA induces TCRαβ(+) CD4(+) CD25(+) FoxP3(+) T regulatory (Treg) cells that suppress Tc1-mediated CS.. Animal model of contact sensitivity was used to study skin-induced suppression.. Current work employing Tc1-mediated CS shows that skin-induced suppression is dose-dependent and declines with time. Experiments with the four non-cross-reacting antigens 2,4-dinitrophenylated bovine serum albumin (DNP-BSA), ovalbumin (OVA), myelin basic protein (MBP) and immunoglobulins conjugated with oxazolone (OX-Ig) employing models of active suppression, "transfer in" and "transfer out" protocols showed that EC immunization with any tested protein antigen inhibits CS response suggesting lack of antigen-specificity of the investigated phenomenon.. The ease of EC generation of antigen-non-specific regulatory cells may have important implications for designing therapeutic schemes aimed at modulating immune responses.

Topics: Adoptive Transfer; Animals; CD8-Positive T-Lymphocytes; Dermatitis, Allergic Contact; Desensitization, Immunologic; Dinitrophenols; Disease Models, Animal; Female; Haptens; Immune Tolerance; Immunoglobulins; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Myelin Basic Protein; Ovalbumin; Oxazolone; Serum Albumin, Bovine; Skin; T-Lymphocytes, Regulatory; Time Factors

The clarification of cutaneous dendritic cell subset and the role of thymic stromal lymphopoietin (TSLP) signaling in epicutaneous sensitization with protein antigens, as in the development of atopic dermatitis, is a crucial issue.. Because TSLP is highly expressed in the vicinity of Langerhans cells (LCs), we sought to clarify our hypothesis that LCs play an essential role in epicutaneous sensitization with protein antigens through TSLP signaling.. By using Langerin-diphtheria toxin receptor knock-in mice and human Langerin-diphtheria toxin A transgenic mice, we prepared mice deficient in LCs. We also prepared mice deficient in TSLP receptors in LCs by using TSLP receptor-deficient mice with bone marrow chimeric technique. We applied these mice to an ovalbumin (OVA)-induced epicutaneous sensitization model.. Upon the epicutaneous application of OVA, conditional LC depletion attenuated the development of clinical manifestations as well as serum OVA-specific IgE increase, OVA-specific T-cell proliferation, and IL-4 mRNA expression in the draining lymph nodes. Consistently, even in the steady state, permanent LC depletion resulted in decreased serum IgE levels, suggesting that LCs mediate the T(H)2 local environment. In addition, mice deficient in TSLP receptors on LCs abrogated the induction of OVA-specific IgE levels upon epicutaneous OVA sensitization.. LCs initiate epicutaneous sensitization with protein antigens and induce T(H)2-type immune responses via TSLP signaling.

Topics: Administration, Cutaneous; Allergens; Animals; Bone Marrow Cells; Chemokines; Chimerism; Cytokines; Dendritic Cells; Dermatitis, Allergic Contact; Disease Models, Animal; Epitopes; Female; Immunoglobulin E; Langerhans Cells; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; OX40 Ligand; Receptors, Cytokine; Signal Transduction; Th2 Cells; Thymic Stromal Lymphopoietin; Up-Regulation

Fenugreek (Trigonella foenum-graecum L.) has a wide variety of therapeutic properties for allergic and inflammatory diseases and is used as a traditional functional food, but its antiallergenic mechanism in these diseases is yet to be clearly elucidated.. In the present study, we investigated the antiallergic activity of fenugreek extract using trimellitic anhydride (TMA)-induced contact hypersensitivity (CHS) mice in vivo and ovalbumin (OVA)-immunized BALB/c mice ex vivo as represented model of T-helper (Th) 2-induced allergy.. BALB/c mice were administered 250 mg/kg body weight (BW) of fenugreek extract for 7 days after sensitization and challenge treatment with 2-5% TMA. Ear thickness were noted, and the infiltration of eosinophils and mast cells was investigated by hematoxylin and eosin (H&E) and toluidine blue (TB) staining. The supernatants from homogenized ear and splenocytes were used for cytokine determination using ELISA. In addition, splenocytes from OVA-immunized BALB/c mice were treated with fenugreek extract ex vivo. The levels of cytokines present in the supernatants were determined by ELISA. The mRNA expression of T-box transcription factor 21 gene (T-bet), GATA-binding protein 3 (GATA-3), interferon (IFN)-γ, and interleukin (IL)-4 were evaluated by real-time RT-PCR.. Fenugreek extract was found to reduce ear thickness as well as the infiltration of eosinophils and mast cells. In homogenized ear, the production of IL-4, IL-5, IL-13, and IL-1β was suppressed. To determine the mechanism by which fenugreek extract inhibits allergic skin inflammation, detailed studies were conducted revealing that fenugreek extract prevented differentiation into Th2 cells in the splenocytes of OVA-induced allergic mice, resulting from suppressing the secretion of IL-4 and mRNA expression of GATA-3, an IL-4 transcription factor. In earlier phase, these extracts enhanced the secretion of IFN-γ, the mRNA expression of T-bet, an IFN-γ transcription factor, and the number of IFN-γ-producing CD4(+) T cells.. These results indicate that fenugreek extract cures Th2-induced allergic skin inflammation by enhancing Th1 differentiation. These data suggest that fenugreek extracts may prove to be an useful therapeutic agent on allergic inflammatory diseases as traditional use as well as Th2-mediated allergic response.

Topics: Allergens; Animals; Anti-Allergic Agents; Cell Proliferation; Cytokines; Dermatitis, Allergic Contact; Female; Mice; Mice, Inbred BALB C; Ovalbumin; Phthalic Anhydrides; Phytotherapy; Plant Extracts; Seeds; Spleen; Trigonella

Interaction between the nervous and immune systems greatly contributes to inflammatory disease. In organs at the interface between our body and the environment, the sensory neuropeptide substance P (SP) is one key mediator of an acute local stress response through neurogenic inflammation but may also alter cytokine balance and dendritic cell (DC) function. Using a combined murine allergic inflammation/noise stress model with C57BL/6 mice, we show in this paper that SP--released during repeated stress exposure--has the capacity to markedly attenuate inflammation. In particular, repeated stress exposure prior to allergen sensitization increases DC-nerve fiber contacts, enhances DC migration and maturation, alters cytokine balance, and increases levels of IL-2 and T regulatory cell numbers in local lymph nodes and inflamed tissue in a neurokinin 1-SP-receptor (neurokinin-1 receptor)-dependent manner. Concordantly, allergic inflammation is significantly reduced after repeated stress exposure. We conclude that SP/repeated stress prior to immune activation acts protolerogenically and thereby beneficially in inflammation.

Topics: Allergens; Animals; Antigen Presentation; Cells, Cultured; Coculture Techniques; Dendritic Cells; Dermatitis, Allergic Contact; Disease Models, Animal; Electric Stimulation; Female; Inflammation Mediators; Langerhans Cells; Mice; Mice, Inbred C57BL; Noise; Ovalbumin; Pilot Projects; Random Allocation; Stress, Physiological; Substance P

Macrophage migration inhibitory factor (MIF) is a pluripotent cytokine that has an essential role in the pathophysiology of experimental allergic inflammation. Recent findings suggest that MIF is involved in several allergic disorders, including atopic dermatitis (AD). In this study, the role of MIF in allergic skin inflammation was examined using a murine model of AD elicited by epicutaneous sensitization with ovalbumin (OVA). We observed the number of skin-infiltrating eosinophils to significantly increase in OVA-sensitized MIF transgenic (Tg) mice compared with their wild-type (WT) littermates. On the other hand, eosinophils were virtually absent from the skin of MIF knockout (KO) mice and failed to infiltrate their skin after repeated epicutaneous sensitization with OVA. The mRNA expression levels of eotaxin and IL-5 were significantly increased in OVA-sensitized skin sites of MIF Tg mice, but were significantly decreased in MIF KO mice in comparison with the levels in WT littermates. Eotaxin expression was induced by IL-4 stimulation in fibroblasts in MIF Tg mice, but not in MIF KO mice. These findings indicate that MIF can induce eosinophil accumulation in the skin. Therefore, the targeted inhibition of MIF might be a promising new therapeutic strategy for allergic skin diseases.

Topics: Animals; Bone Marrow Cells; Cells, Cultured; Chemokine CCL11; Dermatitis, Allergic Contact; Dermis; Disease Models, Animal; Eosinophils; Fibroblasts; Interleukin-5; Intramolecular Oxidoreductases; Macrophage Migration-Inhibitory Factors; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; RNA, Messenger; Th2 Cells

Orally administered milk fat enriched in conjugated linoleic acid (CLA) and trans-vaccenic acid (VA) ('enriched milk fat'), produced by supplementing the diet of pasture-fed cows with fish and sunflower oil, has been shown previously to suppress the development of allergic airway disease in mice.. To investigate whether topical or oral application of enriched milk fat and its two major fatty acids cis-9, trans-11 CLA (c9,t11-CLA) and VA inhibit allergic dermatitis in mice.. Allergic dermatitis was induced in C57BL/6 mice by epicutaneous sensitization of tape-stripped skin with ovalbumin (OVA). Enriched milk fat and its two major fatty acids were either topically applied to the OVA-sensitized skin, or orally fed to mice by supplementation of the diet. Blood and skin tissues were collected for analysis after the third skin sensitization.. Both topical and oral administration of enriched milk fat and its two major fatty acids led to significant suppression of allergic dermatitis as evidenced by reduced clinical and histological scores of affected skins, infiltration of inflammatory cells, and circulating allergen-specific IgE levels, compared with treatment with normal milk fat or the base control diet. C9,t11-CLA and VA individually inhibited multiple facets of allergic dermatitis when topically applied, and their combination produced a strong additive effect.. Enriched milk fat, and its two major fatty acids c9,t11-CLA and vaccenic acid attenuate allergic dermatitis in mice.

Topics: Animals; Cattle; Dermatitis, Allergic Contact; Dietary Supplements; Fats; Female; Linoleic Acids; Mice; Mice, Inbred C57BL; Milk; Oleic Acids; Ovalbumin; Skin Tests

Patients with atopic dermatitis (AD) often suffer from food allergy and develop flares upon skin contact with food allergens. However, it is unclear whether T cells sensitized to allergens in the gut promote this skin inflammation. To address this question, we orally immunized WT mice and mice lacking the skin-homing chemokine receptor Ccr4 (Ccr4-/- mice) with OVA and then challenged them epicutaneously with antigen. Allergic skin inflammation developed in the WT mice but not in the mutants and was characterized by epidermal thickening, dermal infiltration by eosinophils and CD4+ T cells, and upregulation of Th2 cytokines. T cells purified from mesenteric lymph nodes (MLNs) of orally immunized WT mice transferred allergic skin inflammation to naive recipients cutaneously challenged with antigen, but this effect was lost in T cells purified from Ccr4-/- mice. In addition, the ability of adoptively transferred OVA-activated T cells to home to the skin following cutaneous OVA challenge was ablated in mice that lacked lymph nodes. These results indicate that cutaneous exposure to food antigens can reprogram gut-homing effector T cells in LNs to express skin-homing receptors, eliciting skin lesions upon food allergen contact in orally sensitized AD patients.

Topics: Administration, Cutaneous; Administration, Oral; Adoptive Transfer; Allergens; Animals; Chemotaxis, Leukocyte; Cholera Toxin; Dermatitis, Allergic Contact; Food Hypersensitivity; Immunization; Integrins; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Ovalbumin; Receptors, CCR4; Receptors, Fibroblast Growth Factor; Receptors, Lymphocyte Homing; Sialoglycoproteins; Skin; Specific Pathogen-Free Organisms; T-Lymphocyte Subsets

The prevalence of allergies has been linked to Western life style factors including a decrease of microbial exposure. Probiotics, such as Escherichia coli Nissle 1917 (EcN), have been shown to be beneficial for prevention and treatment of several chronic inflammatory diseases.. The aim of this study was to investigate the impact of oral EcN administration on development and outcome of allergen-induced dermatitis.. In sensitized BALB/c mice, skin inflammation was induced by topical allergen application. EcN was administered orally in a preventive manner. Severity of dermatitis was analysed by evaluation of skin score, local cellular and cytokine profile. The systemic immune response was assessed by analysis of immunoglobulins and allergen-dependent cytokine response.. Oral EcN administration improved allergen-induced dermatitis dose-dependently. In parallel, a reduction of epidermal thickness and infiltrating immune cells together with an enhanced number of forkhead box P3 (Foxp3)(+) cells and a trend of increased IFNγ, IL-10 and TGFβ expression was detected in eczematous skin. In allergen-stimulated splenocytes reduced IL-4 and IFNγ along with an elevated IL-10 production and a tendency to an increased TGFβ secretion were observed.. Our findings indicate that EcN alters the local allergen-induced immune response by increase of Foxp3(+) cells and by favouring an immunoregulatory cytokine pattern. Thus, oral administration of EcN might be an effective strategy in prevention and potentially therapy of allergic inflammatory skin diseases.

Topics: Administration, Oral; Allergens; Animals; Cytokines; Dermatitis, Allergic Contact; Disease Models, Animal; Escherichia coli; Female; Forkhead Transcription Factors; Gene Expression; Immunoglobulin A; Mice; Mice, Inbred BALB C; Ovalbumin; Probiotics; RNA, Messenger; T-Lymphocytes, Regulatory

Epidemiological studies suggest that obesity is associated with the impairment of immunity. However, there is no experimental evidence that obesity prejudices immune responses.. This study was designed to determine the effects of obesity on contact hypersensitivity (CHS) response using a diet-induced obese (DIO) mouse model.. The effect of high fat diet (HFD) on CHS response to trinitrochlorobenzene (TNCB) was assessed by ear swelling, cytokine production, functional analysis of epidermal Langerhans cells, and adoptive transfer of immune cells. Immune response to ovalbumin was also analyzed in DIO mice.. C57BL/6 mice but not BALB/c mice that fed with HFD for 4 weeks or more became obese and showed impaired CHS response, although both strain of mice showed enhanced irritant response to TNCB. CHS response was slightly impaired when C57BL/6 mice fed with HFD for 1 or 2 weeks. This suggests that diet-induced obesity or the HFD itself impairs the CHS response in the susceptible mice. The adoptive transfer of immune cells from DIO mice sensitized with TNCB to naïve mice failed to show vigorous CHS, which suggests dysfunction of an afferent phase of CHS in DIO mice. However, the number and allo-stimulating ability of epidermal Langerhans cells were comparable between DIO mice and lean mice. In addition, the immune response to ovalbumin (delayed type hypersensitivity, and antigen-dependent production of antibodies and cytokine) was preserved in DIO mice.. These results suggest that the diet-induced obesity or the HFD only partially impairs immunity in the susceptible mice.

Topics: Adoptive Transfer; Animals; Cytokines; Dermatitis, Allergic Contact; Dietary Fats; Disease Models, Animal; Edema; Female; Hypersensitivity, Delayed; Langerhans Cells; Leptin; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Obesity; Ovalbumin; Picryl Chloride; Species Specificity; T-Lymphocytes; Time Factors

CD44 and l-selectin (CD62L) are major adhesion receptors that mediate leucocyte recruitment at inflammatory sites and lymph nodes, by supporting cell rolling under blood flow. Both CD44 and CD62L have been implicated in inflammatory skin disorders, but their specific involvement in an immediate-type allergic reaction remains uncertain. We used mice deficient in CD44 or CED62L or both in order to determine whether one or both of these molecules were required for leucocyte extravasation in an atopic dermatitis-like allergic response. Wild-type (WT) mice and mice deficient in CD44, CD62L or both were immunized with ovalbumin (OVA). Inflammatory reaction in the ear was elicited once by means of intradermal injection of OVA. Effective sensitization of CD62L knockout (KO) mice required intraperitoneal antigen injection; however, OVA-specific T helper 2 (Th2)-type immune responses and IgE production in mice lacking CD44, CD62L or both were comparable to those in WT mice following intraperitoneal immunization. We employed intravital videomicroscopy to monitor the recruitment of fluorescence-labelled leucocytes to the ear tissue following challenge with OVA. The number of adherent leucocytes was significantly reduced in CD44 KO and CD44/CD62L double KO mice, indicating that CD44 was involved in firm adhesion, the committed step of leucocyte extravasation. Histology of the OVA-challenged ears showed a diminished leucocyte infiltration in the ears of CD44 KO and double KO mice. The results of our study demonstrate that CD44, but not CD62L, is required for leucocyte extravasation during a Th2-type inflammatory response.

Topics: Animals; Cell Movement; Dermatitis, Allergic Contact; Disease Models, Animal; Genotype; Hyaluronan Receptors; Immunoglobulin E; Immunoglobulin G; Immunohistochemistry; Inflammation; Interferon-gamma; Interleukin-4; Kinetics; L-Selectin; Leukocytes; Lymphocytes; Mice; Mice, Knockout; Ovalbumin; Skin; Th2 Cells; Time Factors

Late phase allergic response has been implicated in the pathogenesis of allergic diseases. In the current study, we investigated the role of IL-4, IL-5 and mast cells in the development of cutaneous late phase reaction (LPR) in mice. Antigenic challenge of ears of ovalbumin (OVA)-immunized BALB/c mice caused a biphasic ear swelling peaking at 1 hr (immediate phase reaction; IPR) and 24 hr (LPR). Ear swelling in LPR was significantly suppressed by the treatment with anti-IL-4 monoclonal antibody (mAb) before antigen challenge. Local eosinophil accumulation during LPR, however, was not inhibited by anti-IL-4 mAb. Moreover, anti-IL-5 mAb had no effect on the swelling response though it significantly suppressed the local accumulation of eosinophils. Interestingly, mast cell-deficient mice (WBB6F1-W/Wv) developed LPR without exhibiting IPR, while the magnitude of ear swelling and local eosinophilia was significantly lower than in normal congenic mice (+/+ mice). The present findings show that IL-4 and IL-5 differently regulate the development of LPR, and that IgE-mediated mast cell activation is required for full response.

Topics: Animals; Antibodies, Monoclonal; Cell Movement; Dermatitis, Allergic Contact; Ear, External; Edema; Eosinophils; Hypersensitivity, Delayed; Immunoglobulin E; Interleukin-4; Interleukin-5; Male; Mast Cells; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin

An allergic dermatitis model was developed by repeated sensitization and challenge with antigen (ovalbumin, OA) over 7 months in mice. ddY mice were sensitized by i.p. injection of OA adsorbed on Al(OH)3 (1 microg OA/2 mg Al(OH)3/animal) once every 3 weeks. Antigen challenge was conducted by injection of OA solution (0.1, 1 and 10 microg/site) into the skin of the hind paw instep 10 d after the respective sensitizations. At the 1st challenge, all the 3 groups showed an immediate edematous response with the peak at 30 min or 1 h after the challenge. The group challenged with the highest dose (10 microg/site) of the antigen developed a clear late-phase edema, which was observed at the 2nd challenge, increasing until the 3rd challenge, reaching a plateau at further challenges. On the other hand, such late phase edema scarcely developed in the group challenged with the lowest dose (0.1 microg/site) of the antigen. The amount of circulating specific IgE antibody increased following repeated sensitizations and challenges in all groups, but there were no significant differences in the levels among them. Mepyramine suppressed the early edema by approximately 50%, yet the late phase edema was unaffected. In conclusion, using Al(OH)3+antigen for sensitization and an appropriate amount of antigen for challenge, reproducible biphasic edematous responses were observed long-term without desensitization. This model may be classified as an acute allergic dermatitis and can be useful for quantitatively evaluating the effects of anti-allergic drugs.

Topics: Animals; Antigens; Benzoquinones; Cyclooxygenase Inhibitors; Dermatitis, Allergic Contact; Histamine H1 Antagonists; Indomethacin; Lipoxygenase Inhibitors; Male; Mice; Ovalbumin; Pyrilamine; Reproducibility of Results

We examined the effect of Y-24180, a potent antagonist to platelet-activating factor (PAF), on allergic cutaneous reactions in actively sensitized mice.. Male BALB/c and BALB/c-nu/nu mice were used.. Y-24180, ketotifen fumarate (ketotifen), and suplatast tosilate (suplatast) were orally administered twice a day for 3 days beginning 2 days before an ovalbumin (OA) challenge. Hydrocortisone 17-butyrate (hydrocortisone) was applied topically to ear surface once a day for 3 days, beginning 2 days before the OA challenge.. Mice actively sensitized with OA were challenged by intradermally injecting OA into both ears. Ear thickness was measured with a dial thickness gauge.. Increase in ear thickness, with peak responses at 1 h (immediate phase reaction, IPR) and 24 h (late phase reaction, LPR) after the challenge, were induced in actively sensitized BALB/c mice. The reactions were not induced in T cell-deficient BALB/c-nu/nu mice. Y-24180 suppressed both the IPR and LPR of BALB/c mice. Although suplatast suppressed the LPR, the IPR was not affected. Ketotifen suppressed the IPR, but not the LPR. Hydrocortisone suppressed both the IPR and LPR of BALB/c mice. Furthermore, Y-24180 in combination with hydrocortisone significantly enhanced the effect of hydrocortisone on both the reactions.. Y-24180 was demonstrated not only to suppress the IPR and LPR, but also to show strong suppressive effects in combination with topical hydrocortisone. Therefore, Y-24180 is expected to contribute to the treatment of inflammatory skin diseases including atopic dermatitis.

Topics: Administration, Oral; Administration, Topical; Allergens; Animals; Anti-Allergic Agents; Anti-Inflammatory Agents; Arylsulfonates; Azepines; Dermatitis, Allergic Contact; Ear; Eosinophils; Histamine H1 Antagonists; Hydrocortisone; Immunoglobulin E; Immunoglobulin G; Ketotifen; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Ovalbumin; Platelet Membrane Glycoproteins; Receptors, Cell Surface; Receptors, G-Protein-Coupled; Sulfonium Compounds; Time Factors; Triazoles

Dendritic cells (DCs) are special subsets of antigen-presenting cells characterized by their highly potent capacity to activate immunologically naive T cells. Here we report that DCs that are transfected with CD95 ligand (CD95L) cDNA, called 'killer' DCs, deliver death signals, instead of activation signals, to T cells after antigen-specific interaction. Injection of antigen-pulsed killer DCs into mice before sensitization induced antigen-specific immunosuppression. When administered after sensitization, killer DCs suppressed immune responses almost completely after subsequent challenge. Thus, killer DCs represent an entirely new immunomodulatory protocol, which may become directly applicable in preventing and even treating T cell-mediated inflammatory diseases.

Topics: Animals; Antibodies, Monoclonal; Apoptosis; CD4-Positive T-Lymphocytes; Cell Line; Cell Transplantation; Clone Cells; Dendritic Cells; Dermatitis, Allergic Contact; Dinitrofluorobenzene; DNA, Complementary; Fas Ligand Protein; fas Receptor; Female; Hypersensitivity, Delayed; Immune Tolerance; Immunity, Cellular; Liver; Membrane Glycoproteins; Mice; Ovalbumin; Transfection

Recently we demonstrated that activated rat macrophages produced neutrophil chemotactic factors (chemokines) including cytokine-induced neutrophil chemoattractant (CINC)-1, CINC-2alpha, CINC-2beta, CINC-3/rat macrophage inflammatory protein (MIP)-2 and rat MIP-1alpha (rMIP-1alpha).. In the present study, by using an enzyme-linked immunosorbent assay specific for each chemokine, we determined the levels of the chemokines in the pouch fluid (inflammatory site) of the fluorescein isothiocyanate-labeled ovalbumin (FITC-OVA)-induced allergic inflammation in rats. Effects of anti-chemokine antibodies on neutrophil chemotaxis were determined in vivo and in vitro.. CINC-1 was the major chemokine which rapidly increased after challenge with FITC-OVA, whereas CINC-3 was a minor one, and CINC-2, CINC-3 and rMIP-1alpha increased slowly with a lag time of about 2 h. Anti-CINC-1/CINC-2 antibodies, which inhibited all the CINCs, suppressed both neutrophil infiltration in vivo and neutrophil chemotactic activity of the 8-hour pouch fluid in vitro, whereas anti-rMIP-1alpha antibody slightly suppressed the chemotaxis in vivo and in vitro.. Our results suggest that CINCs, especially CINC-1 and CINC-2, play an important role in the infiltration of neutrophils into the inflammatory site of FITC-OVA-induced allergic inflammation in rats.

Topics: Animals; Body Fluids; Chemokines; Chemotaxis, Leukocyte; Dermatitis, Allergic Contact; Enzyme-Linked Immunosorbent Assay; Immunization; Inflammation; Leukocyte Count; Male; Neutrophils; Ovalbumin; Rabbits; Rats; Rats, Wistar

The effects of nebulized IFN-gamma on primary and secondary IgE production and development of airway hyper-responsiveness (AHR) were investigated. BALB/c mice received primary exposure to aerosolized OVA daily for 10 days and developed anti-OVA IgE responses, immediate cutaneous reactivity to OVA, and altered airway function when assayed on day 12. After secondary exposure to OVA challenges on days 30 and 31, these mice developed an amplified IgE response, heightened cutaneous reactivity to OVA and AHR when measured on day 37. Administration of IFN-gamma for 13 days, beginning 3 days prior to and during primary OVA sensitization, resulted in a decrease in anti-OVA IgE, increases in serum anti-OVA IgG2a levels, a decrease in cutaneous reactivity to OVA, and normal airway function when assessed on day 12 after primary sensitization. This treatment also prevented the development of secondary anti-OVA IgE responses and altered airway responsiveness but did not induce a secondary rise in anti-OVA IgG2a in the serum measured on day 37. Treatment with IFN-gamma on days 26 to 30, well after primary responses were established but just prior to secondary OVA challenge, abolished the development of secondary anti-OVA IgE responses, resulted in an increase in anti-OVA IgG2a in the serum, and prevented the development of AHR. In vitro, CD4+ T cells obtained from OVA-sensitized mice treated with either "early" or "late" IFN-gamma inhibited IgE production. Delivery of IFN-gamma to the airways can prevent secondary allergen sensitization even after primary sensitization has been achieved and this effect is mediated by CD4+ T cells.

Topics: Administration, Inhalation; Aerosols; Animals; CD4-Positive T-Lymphocytes; Depression, Chemical; Dermatitis, Allergic Contact; Immunization; Immunoglobulin Class Switching; Immunoglobulin E; Immunoglobulin G; Immunologic Factors; Immunologic Memory; Interferon-gamma; Lymphocyte Activation; Lymphocyte Cooperation; Mice; Mice, Inbred BALB C; Models, Immunological; Nebulizers and Vaporizers; Ovalbumin; Respiratory Hypersensitivity; Time Factors

To investigate the mechanisms underlying airway hyperresponsiveness a murine model was developed with several important characteristics of human allergic asthma. Mice were intraperitoneally sensitized with ovalbumin and after 4 weeks challenge via an ovalbumin aerosol. After aerosol, lung function was evaluated with a non-invasive forced oscillation technique. The amount of mucosal exudation into the airway lumen and the presence of mast cell degranulation was determined. Tracheal responsiveness was measured at several time points after challenge. At these time points also bronchoalveolar lavage and histology were performed. Sensitization induced high antigen-specific IgE levels in serum. Inhalation of ovalbumin in sensitized mice induced an immediate but no late bronchoconstrictive response. During this immediate phase, respiratory resistance was increased (54%). Within the first hour after ovalbumin inhalation increased mucosal exudation and mast cell degranulation were observed. At 12 and 24 h after ovalbumin challenge, mice showed tracheal hyperresponsiveness (29% and 34%, respectively). However, no apparent inflammation was found in the lungs or bronchoalveolar lavage. From these results it can be concluded that hyperresponsiveness can develop via mechanisms independent of an inflammatory infiltrate. Since mast cell degranulation occurred after ovalbumin exposure, we hypothesize that mast cells are involved in the induction of airway hyperresponsiveness in this model.

Topics: Animals; Bronchial Hyperreactivity; Bronchoalveolar Lavage; Bronchoconstriction; Dermatitis, Allergic Contact; Exudates and Transudates; Immunoglobulin E; Immunoglobulin G; In Vitro Techniques; Lung; Male; Methacholine Chloride; Mice; Mice, Inbred BALB C; Ovalbumin; Respiratory Hypersensitivity; Trachea