oxazolone and Hypersensitivity

Epidemiological studies have long reported that perturbations of the childhood microbiome increase the risk of developing allergies, but a causal relationship with atopic dermatitis remains unclear. Here we colonized germ-free mice at birth or at one or eight week-of-age to investigate the role of prenatal and early postnatal microbial exposure on development of oxozolone-induced dermatitis later in life. We demonstrate that only one week delayed microbial colonization increased IgE levels and the total histological score of the inflamed ear compared to mice colonized throughout life. In parallel, several pro-inflammatory cytokines and chemokines were upregulated in the ear tissue demonstrating an enhanced immunological response following delayed postnatal colonization of the gut. In contrast, sensitivity to oxazolone-induced dermatitis was unaffected by the presence of a maternal microbiota during gestation. Mice colonized at eight week-of-age failed to colonize Rikenellaceae, a group of bacteria previously associated with a high-responding phenotype, and did not develop an immunological response to the same extent as the early colonized mice despite pronounced histopathological manifestations. The study provides proof-of-principle that the first intestinal colonizers of mice pups are crucial for the development of oxazolone-induced dermatitis later in life, and that the status of the maternal microbiota during pregnancy has no influence on the offspring's allergic immune response. This highlights an important window of opportunity following birth for microbiota-mediated interventions to prevent atopic responses later in life. How long such a window is open may vary between mice and humans considering species differences in the ontogeny of the immune system.

Topics: Animals; Bacteria; Cytokines; Dermatitis, Atopic; Disease Models, Animal; Dysbiosis; Female; Gastrointestinal Microbiome; Humans; Hypersensitivity; Immunoglobulin E; Mice; Oxazolone; Pregnancy; Prenatal Exposure Delayed Effects; RNA, Ribosomal, 16S

Hapten contact hypersensitivity (CHS) elicits a well-documented inflammation response that can be used to illustrate training of immune cells through hapten-specific CHS memory. The education of hapten-specific memory T cells has been well-established, recent research in mice has expanded the "adaptive" characteristic of a memory response from solely a function of the adaptive immune system, to innate cells as well. To test whether similar responses are seen in a non-rodent model, we used hapten-specific CHS to measure the ear inflammation response of outbred pigs to dinitrofluorobenzene (DNFB), oxazolone (OXA), or vehicle controls. We adapted mouse innate memory literature protocols to the domestic pig model. Animals were challenged up to 32 days post initial sensitization exposure to the hapten, and specific ear swelling responses to this challenge were significant for 7, 21, and 32 days post-sensitization. We established hapten-specific CHS memory exists in a non-rodent model. We also developed a successful protocol for demonstrating these CHS responses in a porcine system.

Topics: Adjuvants, Immunologic; Animals; Dinitrofluorobenzene; Disease Models, Animal; Female; Haptens; Hypersensitivity; Immunologic Memory; Male; Otitis; Oxazolone; Swine

Filaggrin (Flg) and hornerin (Hrnr) share similar structural and functional features. Both proteins have been implicated as essential proteins for skin barrier maintenance. Loss-of-function mutations of these genes constitute a risk factor for atopic dermatitis and eczema-related asthma. Furthermore, both FLG and HRNR protein levels are downregulated in patients with atopic dermatitis. Thus, mice deficient for Flg and Hrnr provide a novel model to study skin barrier impairment and the susceptibility for cutaneous inflammation.. By using appropriate targeting vectors and breeding strategies, we established a homozygous FlgHrnr double-deficient (FlgHrnr. Neonates appeared normal, but developed a transient scaly phenotype with overall flaky appearance, but no overt skin phenotype in adulthood, thereby reflecting a subclinical barrier defect seen in humans. Structurally, FlgHrnr. Together, our FlgHrnr

Topics: Adaptive Immunity; Animals; Biopsy; Calcium-Binding Proteins; Disease Models, Animal; Epidermis; Filaggrin Proteins; Hypersensitivity; Immunity, Innate; Immunohistochemistry; Intermediate Filament Proteins; Mice; Mice, Knockout; Oxazolone; Permeability; Phenotype

Constantly increasing prevalence of allergic diseases determines the attempts to elaborate the therapeutic strategies activating immune tolerance to particular allergen. Our current research focuses on the antigen-specific action of CD8+ suppressor T (Ts) lymphocytes induced in mice by intravenous administration of a high dose of haptenated syngeneic erythrocytes. While the regulatory activity of Ts cells mediated by exosome-delivered miRNA-150 is well de ned, the mechanism of their induction remained unclear. Therefore, the current studies investigated the immune e ects induced in mice by intravenous administration of contact allergens coupled to syngeneic erythrocytes. In mouse models of hapten-induced contact hypersensitivity (CHS) and delayed-type hypersensitivity to ovalbumin, we have shown that intravenous administration of hapten-coupled erythrocytes failed to induce CHS effector cells. Moreover, hapten-induced CHS reaction occurred to be suppressed in mice intravenously administered with syngeneic erythrocytes coupled with protein allergen. Finally, we have demonstrated that intravenously administered allergen induces immune tolerance only when bound to syngeneic erythrocytes, proving that intravenously delivered allergens are deprived of their immunizing properties when coupled with membrane of self cells. Altogether, our current studies suggest that alteration of self cell membrane by allergen binding is enough to induce Ts cell-mediated immune tolerance to nonpathogenic agents, which express a great translational potential in such conditions as allergies and hypersensitivity-related autoimmune disorders.

Topics: Allergens; Animals; Dermatitis, Contact; Erythrocyte Transfusion; Haptens; Hypersensitivity; Immune Tolerance; Mice; Mice, Inbred CBA; Oxazolone; T-Lymphocyte Subsets; Transplantation, Isogeneic; Trinitrobenzenes

Upon antigen/allergen recognition, epidermal Langerhans' cells (LC) are mobilized and migrate to the local lymph node where they play a major role in initiating or regulating immune responses. It had been proposed that all chemical allergens induce LC migration via common cytokine signals delivered by TNF-α and IL-1β. Here the dependence of LC migration on TNF-α following treatment of mice with various chemical allergens has been investigated. It was found that under standard conditions the allergens oxazolone, paraphenylene diamine, and trimellitic anhydride, in addition to the skin irritant sodium lauryl sulfate, were unable to trigger LC mobilization in the absence of TNF-α signalling. In contrast, two members of the dinitrohalobenezene family (2,4-dinitrochlorobenzene [DNCB] and 2,4-dinitrofluorobenzene [DNFB]) promoted LC migration independently of TNF-R2 (the sole TNF-α receptor expressed by LC) and TNF-α although the presence of IL-1β was still required. However, increasing doses of oxazolone overcame the requirement of TNF-α for LC mobilization, whereas lower doses of DNCB were still able to induce LC migration in a TNF-α-independent manner. These novel findings demonstrate unexpected heterogeneity among chemical allergens and furthermore that LC can be induced to migrate from the epidermis via different mechanisms that are either dependent or independent of TNF-α. Although the exact mechanisms with regard to the signals that activate LC have yet to be elucidated, these differences may translate into functional speciation that will likely impact on the extent and quality of allergic sensitization.

Topics: Adjuvants, Immunologic; Allergens; Animals; Cell Movement; Dinitrofluorobenzene; Epidermis; Hypersensitivity; Immunization; Langerhans Cells; Mice; Mice, Inbred BALB C; Mice, Knockout; Oxazolone; Receptors, Tumor Necrosis Factor, Type II; Signal Transduction; Tumor Necrosis Factor-alpha

IL-10 plays a critical role in the induction of specific T-cell tolerance. To date, whether IL-10 induction by antigen application is dose- or time-dependent remains unclear. In this study, IL-10 induction by allergen exposure was investigated in the several schedules. Oxazolone was repeatedly applied to mouse ear, and mRNA of inflammatory cytokines in lesional skins was measured. The results indicated that continuous high-dose antigen exposure induces IL-4 as well as abundant IL-10 production. Monocytes/dendritic cells and T cells are major source of IL-10. Allergen-specific immunotherapy is resumed before antigen scattering: preseason. We evaluated safe-loading dose of allergens in preseasonal therapy focusing Tr1 induction. Restarting immunotherapy with high dose effectively augmented IL-10 expression accompanied with further induction of IL-4 and inflammatory cytokines. Therefore, the protocol restarting with low-dose antigen is preferential to obviate the risk of exacerbation or anaphylaxis.

Topics: Anaphylaxis; Animals; Antigens; Desensitization, Immunologic; Humans; Hypersensitivity; Interleukin-10; Interleukin-4; Lymph Nodes; Mice; Models, Animal; Oxazolone; RNA, Messenger; Spleen; T-Lymphocytes, Regulatory

A recent paper suggests that reduced exposure to germs results in the expansion of a cell type called natural killer T cells, which predisposes to colitis and asthma. Such a scenario could explain the Hygiene Hypothesis, which has been a puzzle for decades.

Topics: Animals; Asthma; Cell Proliferation; Colitis; Environmental Exposure; Humans; Hygiene Hypothesis; Hypersensitivity; Interleukins; Metagenome; Mice; Natural Killer T-Cells; Oxazolone; Specific Pathogen-Free Organisms

Neonates have a developing immune response, with a predisposition towards induction of tolerance. As the immune system develops, immunity rather than tolerance is induced, with this development of immunity occurring in response to external factors such as the environment. As ultraviolet radiation (UVR) suppresses immunity, it is likely that the effect of UVR on the neonatal immune system would be augmentation of the suppressive response. In support, childhood exposure to UVR has been linked with an increased incidence of melanoma; consistent with an increase in suppression. To address this, phenotypic and functional immune system studies were undertaken at 8 weeks after one single exposure of solar-simulated UVR to mice, when mice had reached adulthood. Subtle changes were observed in cell populations resident in the skin-draining lymph nodes (LNs) and there also appeared to be a subtle, but not statistically significant, increase in the production of interleukin-10 and interferon-γ. Importantly, these changes also corresponded with significant suppression of the contact hypersensitivity response in irradiated mice compared with their control counterparts. This suppression was apparent when antigen sensitisation occurred during the neonatal or adult period, and thus did not appear to be analogous to UVR-induced suppression in adults. Although the percentage of T regulatory cells was increased in the skin-draining LNs, they were induced in a different manner to those induced following adult UVR exposure, with no increase in function on a per-cell basis. It therefore appears that one single neonatal exposure to UVR alters development of the immune system, leading to long-term implications for induction of immunity.

Topics: Animals; Animals, Newborn; Cell Proliferation; Cells, Cultured; Dermatitis, Contact; Female; Hypersensitivity; Immune System; Immune Tolerance; Interferon-gamma; Interleukin-10; Lymph Nodes; Lymphocyte Activation; Male; Mice; Mice, Inbred BALB C; Oxazolone; Skin; T-Lymphocytes, Regulatory; Ultraviolet Rays

We set out to establish the in vivo histamine H(1) receptor antagonistic (antihistaminic) and antiallergic properties of bilastine.. In vivo antihistaminic activity experiments consisted of measurement of: inhibition of increase in capillary permeability and reduction in microvascular extravasation and bronchospasm in rats and guinea pigs induced by histamine and other inflammatory mediators; and protection against lethality induced by histamine and other inflammatory mediators in rats. In vivo antiallergic activity experiments consisted of measurement of passive and active cutaneous anaphylactic reactions as well as type III and type IV allergic reactions in sensitised rodents.. In the in vivo antihistaminic activity experiments, bilastine was shown to have a positive effect, similar to that of cetirizine and more potent than that of fexofenadine. The results of the in vivo antiallergic activity experiments showed that the properties of bilastine in this setting are similar to those observed for cetirizine and superior to fexofenadine in the model of passive cutaneous anaphylactic reaction. When active cutaneous anaphylactic reaction experiments were conducted, bilastine showed significant activity, less potent than that observed with cetirizine but superior to that of fexofenadine. Evaluation of the type III allergic reaction showed that of the antihistamines only bilastine was able to inhibit oedema in sensitised mice, although its effect in this respect was much less potent than that observed with dexamethasone. In terms of the type IV allergic reaction, neither bilastine, cetirizine nor fexofenadine significantly modified the effect caused by oxazolone.. The results of our in vivo preclinical studies corroborate those obtained from previously conducted in vitro experiments of bilastine, and provide evidence that bilastine possesses antihistaminic as well as antiallergic properties, with similar potency to cetirizine and superior potency to fexofenadine.

Topics: Animals; Benzimidazoles; Bradykinin; Bronchial Spasm; Capillary Permeability; Cetirizine; Dermatitis; Dose-Response Relationship, Drug; Guinea Pigs; Histamine; Histamine Antagonists; Hypersensitivity; Male; Mice; Molecular Structure; Oxazolone; Piperidines; Rats; Rats, Wistar; Receptors, Histamine H1; Serotonin; Structure-Activity Relationship; Terfenadine; Time Factors

CD1d-restricted T-cells are activated by glycolipids presented by the major histocompatibility complex class-Ib molecule CD1d, found on the surface of antigen-presenting cells (APC). This interaction between APC, most notably dendritic cells (DC), and CD1d-restricted T-cells is an important regulatory step in the initiation of adaptive immune responses. It is well known that DC play a crucial role in the induction of contact hypersensitivity (CHS), a frequently studied form of in vivo T-cell-mediated immunity. In this study, we show that CD1d-restricted T-cells are also necessary for CHS, because both wild-type mice treated systemically or topically with CD1d glycolipid antagonists and CD1d-restricted T-cell-null mice have markedly diminished CHS responses. Thus, pharmacologic antagonists of CD1d can be used as effective inhibitors of CHS, a prototype for a variety of delayed-type tissue hypersensitivity responses.

Topics: Administration, Topical; Animals; Antigen Presentation; Antigens, CD1; Antigens, CD1d; Cell Line; Dendritic Cells; Dermatitis; Dermatitis, Contact; Dose-Response Relationship, Drug; Glycolipids; Hypersensitivity; Killer Cells, Natural; Major Histocompatibility Complex; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Transgenic; Mutation; Oxazolone; Phosphatidylethanolamines; Polyethylene Glycols; Receptors, Antigen, T-Cell

The objective of the present study was to investigate the immunomodulatory activity of Argyreia speciosa on cellular and humoral immunity. Oral administration of the ethanolic extract of A. speciosa root (ASEE), at the doses of 50, 100 and 200 mg/kg in mice, dose-dependently potentiated the delayed-type hypersensitivity reaction induced both by sheep red blood cells (SRBC) and oxazolone. It significantly enhanced the production of circulating antibody titre in mice in response to SRBC. ASEE failed to show any effect on macrophage phagocytosis. Chronic administration of ASEE significantly ameliorated the total white blood cell count and also restored the myelosuppressive effects induced by cyclophosphamide. The present investigation reveals that ASEE possesses immunomodulatory activity.

Topics: Animals; Antibodies; Blood Cell Count; Convolvulaceae; Cyclophosphamide; Dose-Response Relationship, Drug; Erythrocytes; Female; Hypersensitivity; Hypersensitivity, Delayed; Immunosuppression Therapy; Macrophages; Male; Mice; Oxazolone; Penicillamine; Phagocytosis; Plant Extracts; Plant Roots; Sheep

Chemical-induced allergy continues to be an important occupational health problem. Despite decades of investigation, the molecular mechanisms underlying chemical-induced hypersensitivity and irritancy remain unclear because of the complex interplay between properties of different chemicals and the immune system. In this study, gene expression induced by toluene diisocyanate (TDI, a primarily IgE-inducing sensitizer), oxazolone (OXA, a cell-mediated hypersensitivity inducing sensitizer), or nonanoic acid (NA, a non-sensitizing irritant) was investigated using gene arrays. Female BALB/c mice were dermally exposed on the ears once daily for 4 consecutive days. On day 5, the lymph nodes draining the exposure sites were collected and used for RNA extraction and subsequent hybridization to Affymetrix Mu6500 oligonucleotide arrays. Of the 6519 genes on the arrays, there were 44, 13, and 51 genes in the TDI-, OXA-, and NA-exposed samples, respectively, that displayed a minimum of twofold change in expression level relative to the vehicle control. There were 32, 19, and 19 genes that were differentially expressed (with a minimum of twofold change) between TDI and OXA, TDI and NA, OXA and NA, respectively. The differentially expressed genes include immune response-related genes, transcriptional factors, signal transducing molecules, and Expressed Sequence Tags. Based on the gene array results, candidate genes were further evaluated using RT-PCR. There was only about 47% concordance between the gene array and RT-PCR results.

Topics: Animals; Fatty Acids; Female; Gene Expression; Gene Expression Profiling; Humans; Hypersensitivity; Immunization; Irritants; Mice; Mice, Inbred BALB C; Occupational Diseases; Oligonucleotide Array Sequence Analysis; Oxazolone; Reverse Transcriptase Polymerase Chain Reaction; Toluene 2,4-Diisocyanate

The role of the interferon-gamma (IFN-gamma) receptor 1 (IFN-gammaR1) was investigated in the regulation of MHC expression in kidney in the basal state, in response to potent inflammatory stimuli, and after renal injury. In this study, MHC regulation in mice lacking IFN-gammaR due to targeted disruption of the IFN-gammaR1 gene (GRKO mice) was compared with regulation in 129Sv/J mice with wild-type IFN-gammaR1 genes. Basal class I expression was reduced by approximately 45% in kidneys of GRKO mice, while basal class II expression was confined to interstitial cells and was not reduced in GRKO kidneys. Recombinant IFN-gamma administration induced widespread expression of class I and II in renal tubules, arterial endothelium, and glomeruli of 129Sv/J mice, but produced no change in kidneys of GRKO mice. Potent systemic inflammatory stimuli (injections of allogeneic cells, skin sensitization with oxazolone, and injection of bacterial lipopolysaccharide) significantly induced both class I and class II expression in 129Sv/J mice, but not in GRKO mice. Acute renal injury increased local expression of class I and II in both 129Sv/J and GRKO mice, but the induction in GRKO mice was reduced compared with 129Sv/J mice. Thus, the IFN-gamma receptor plays a unique and nonredundant role in the regulation of renal MHC in the response to inflammation, in the response to renal injury, and in the basal state.

Topics: Adjuvants, Immunologic; Animals; Antigens, Neoplasm; Gene Expression; Gentamicins; Histocompatibility Antigens Class I; Histocompatibility Antigens Class II; Hypersensitivity; Interferon gamma Receptor; Interferon-gamma; Kidney; Kidney Diseases; Lipopolysaccharides; Major Histocompatibility Complex; Mice; Mice, Knockout; Oxazolone; Receptors, Interferon; Recombinant Proteins; Skin

Exposure to synthetic glucocorticoids (GCs) or other stimuli around birth may affect neuroendocrine and immune responsiveness in the offspring. Experiments were conducted to investigate whether maternal manipulation with saline or with GCs alters the corticosterone (CORT) response to a mild stressor in the offspring, and whether maternal manipulation results in long-term altered in vivo humoral and cellular immune responsiveness in the offspring. Pregnant rats were given dexamethasone (DEX, 1.2 mg/kg body weight, i.p.) or saline (SAL) at day 17 and 19 of gestation. A third group of pregnant rats was left undisturbed (UNTR-group). After maternal DEX treatment, no altered CORT response was seen to a novel environment at 20 days of age, as compared to both the SAL-treated group and the UNTR-group. However, saline administration to pregnant rats caused an increased CORT response in female offspring, but not male offspring, as compared to the UNTR-group (P < or = 0.01). Furthermore, no effects of maternal DEX exposure were seen on IgG2a production after immunization with a conjugated pneumococcal polysaccharide (PPS-14-CRM197) at 6 weeks of age. However, maternal SAL treatment enhanced anti-PPS-14 IgG2a antibody levels in female offspring, but not in male offspring, as compared to the UNTR-group (P < or = 0.05). Cellular immune responses were measured by an oxazolone-induced contact hypersensitivity response (CHS-response), at 8 weeks of age. Maternal SAL treatment increased the CHS response in adult male rats, but not in female rats, as compared to both the UNTR-group and the DEX-group (P < or = 0.005). These data suggest that manipulations during late pregnancy not only affect endocrine responsiveness, but also influence immune responsiveness in the rat offspring. Furthermore, these effects may be long-term and gender-specific.

Topics: Adjuvants, Immunologic; Animals; Anti-Inflammatory Agents; Antigens, Bacterial; Bacterial Proteins; Corticosterone; Dexamethasone; Female; Hypersensitivity; Immune System; Immunization; Male; Oxazolone; Pituitary-Adrenal System; Polysaccharides, Bacterial; Pregnancy; Pregnancy, Animal; Rats; Rats, Wistar; Sex Factors; Sodium Chloride; Streptococcus pneumoniae; Stress, Physiological