ovalbumin and Dermatitis--Contact

Langerhans cells (LCs) are antigen-presenting dendritic cells (DCs) that reside in epithelia. The best studied example is the LC of the epidermis. By electron microscopy, their identifying feature is the unique rod- or tennis racket-shaped Birbeck granule. The phenotypic hallmark is their expression of the C-type lectin receptor langerin/CD207. Langerin, however, is also expressed on a recently discovered population of DC in the dermis and other tissues of the body. These 'dermal langerin(+) dendritic cells' are unrelated to LCs. The complex field of langerin-negative dermal DCs is not dealt with here. In this article, we briefly review the history, ontogeny, and homeostasis of LCs. More emphasis is laid on the discussion of functional properties in vivo. Novel models using genetically engineered mice are contributing tremendously to our understanding of the role of LCs in eliciting adaptive immune responses against pathogens or tumors and in inducing and maintaining tolerance against self antigens and innocuous substances in vivo. Also, innate effector functions are increasingly being recognized. Current activities in this area are reviewed, and possibilities for future exploitation of LC in medicine, e.g. for the improvement of vaccines, are contemplated.

Topics: Adaptive Immunity; Animals; Antigen Presentation; Antigens, CD; Antigens, Surface; Cell Lineage; Communicable Diseases; Dermatitis, Contact; Disease Models, Animal; Homeostasis; Humans; Immune Tolerance; Immunity, Innate; Langerhans Cells; Lectins, C-Type; Mannose-Binding Lectins; Mice; Mice, Transgenic; Neoplasms; Ovalbumin; Phenotype; Skin; Vaccines

Topics: Animals; Dermatitis, Contact; Dogs; Drug Eruptions; Graft vs Host Disease; Graft vs Host Reaction; Guinea Pigs; Haptens; Hypersensitivity, Delayed; Kidney Transplantation; Leukocytes; Lymphocytes; Lymphoid Tissue; Macrophages; Microscopy, Electron; Microscopy, Phase-Contrast; Ovalbumin; Plasma Cells; Rabbits; Skin; Skin Transplantation; Transplantation Immunology; Transplantation, Homologous; Tuberculin Test

The histamine H4 receptor (H(4)R) has been shown to have preclinical involvement in both inflammatory and pruritic responses. JNJ-39758979 [(R)-4-(3-amino-pyrrolidin-1-yl)-6-isopropyl-pyrimidin-2-ylamine] is a potent and selective H(4)R antagonist with a Ki at the human receptor of 12.5 ± 2.6 nM and greater than 80-fold selectivity over other histamine receptors. The compound also exhibited excellent selectivity versus other targets. JNJ-39758979 showed dose-dependent activity in models of asthma and dermatitis consistent with other H(4)R antagonists. Preclinical toxicity studies of up to 6 months in rats and 9 months in monkeys indicated an excellent safety profile, supporting the clinical testing of the compound. An oral formulation of JNJ-39758979 was studied in a phase 1 human volunteer study to assess safety, pharmacokinetics, and pharmacodynamics. The compound was well tolerated, with the exception of dose-dependent nausea, and no safety issues were noted in the phase 1 study. JNJ-39758979 exhibited good pharmacokinetics upon oral dosing with a plasma half-life of 124-157 hours after a single oral dose. In addition, dose-dependent inhibition of histamine-induced eosinophil shape change was detected, suggesting that the H4R was inhibited in vivo. In conclusion, JNJ-39758979 is a potent and selective H(4)R antagonist that exhibited good preclinical and phase 1 safety in healthy volunteers with evidence of a pharmacodynamics effect in humans.

Topics: Animals; Anti-Inflammatory Agents; Asthma; Cell Shape; Dermatitis, Contact; Double-Blind Method; Eosinophils; Female; Humans; Isothiocyanates; Macaca fascicularis; Male; Ovalbumin; Pyrimidines; Rats; Receptors, G-Protein-Coupled; Receptors, Histamine; Receptors, Histamine H4; Thiazoles

In vivo histamine release in cutaneous allergic reactions was chronologically examined using the skin microdialysis technique. Antigen challenge was made in ovalbumin-sensitized guinea pigs 1 h after the implantation of the microdialysis probe. A marked histamine release after intradermal injection of ovalbumin solution was observed in the early phase (up to 40 min). No isolated delayed histamine release was observed thereafter over 8 h. Also, in an atopic volunteer, a marked release of histamine after intradermal injection of mite extract was observed only in the early phase, although erythema and induration at the site of injection were continuously observed in the late phase (up to 4 h). These findings suggest that the skin microdialysis technique is a useful, simple technique for chronological determination of chemical mediators released in the allergic skin in vivo.

Topics: Adult; Animals; Dermatitis, Atopic; Dermatitis, Contact; Guinea Pigs; Histamine Release; Humans; Male; Microdialysis; Mites; Ovalbumin; Skin; Time Factors

The interplay between microbes and surface organs, such as the skin, shapes a complex immune system with several checks and balances. The first-line defense is mediated by innate immune pathways leading to inflammation. In the second phase specific T cells invade the infected organ, amplifying inflammation and defense. Consecutively, termination of inflammation is crucial to avoid chronic inflammation triggered by microbes, such as in patients with atopic dermatitis.. We aimed to elucidate how the Staphylococcus aureus-derived cell-wall component lipoteichoic acid (LTA) governs the second phase of immune responses when high concentrations of LTA access T cells directly through disrupted skin.. We analyzed the direct exposure of T cells to LTA in vitro. For in vivo analyses, we used fluorescein isothiocyanate contact hypersensitivity and ovalbumin-induced dermatitis as models for TH2-mediated cutaneous inflammation.. We observed that LTA potently suppressed T-lymphocyte activation in a Toll-like receptor 2-independent manner. LTA-exposed T cells did not proliferate and did not produce cytokines. Importantly, these T cells remained completely viable and were responsive to consecutive activation signals on subsequent removal of LTA. Thus LTA exposure resulted in temporary functional T-cell paralysis. In vivo experiments revealed that T-cell cytokine production and cutaneous recall responses were significantly suppressed by LTA.. We identified a new mechanism through which bacterial compounds directly but temporarily modulate adaptive immune responses.

Topics: Allergens; Animals; Cell Proliferation; Cytokines; Dermatitis, Atopic; Dermatitis, Contact; Fluorescein-5-isothiocyanate; Lipopolysaccharides; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Myeloid Differentiation Factor 88; Ovalbumin; Staphylococcus aureus; T-Lymphocytes; Teichoic Acids; Toll-Like Receptor 2

In this study, we evaluated the effect chronic helminth infection has on allergic disease in mice previously sensitized to OVA. Ten weeks of infection with Litomosoides sigmodontis reduced immunological markers of type I hypersensitivity, including OVA-specific IgE, basophil activation, and mast cell degranulation. Despite these reductions, there was no protection against immediate clinical hypersensitivity following intradermal OVA challenge. However, late-phase ear swelling, due to type III hypersensitivity, was significantly reduced in chronically infected animals. Levels of total IgG2a, OVA-specific IgG2a, and OVA-specific IgG1 were reduced in the setting of infection. These reductions were most likely due to increased Ab catabolism as ELISPOT assays demonstrated that infected animals do not have suppressed Ab production. Ear histology 24 h after challenge showed infected animals have reduced cellular infiltration in the ear, with significant decreases in numbers of neutrophils and macrophages. Consistent with this, infected animals had less neutrophil-specific chemokines CXCL-1 and CXCL-2 in the ear following challenge. Additionally, in vitro stimulation with immune complexes resulted in significantly less CXCL-1 and CXCL-2 production by eosinophils from chronically infected mice. Expression of FcγRI was also significantly reduced on eosinophils from infected animals. These data indicate that chronic filarial infection suppresses eosinophilic responses to Ab-mediated activation and has the potential to be used as a therapeutic for pre-existing hypersensitivity diseases.

Topics: Animals; Antigen-Antibody Complex; Basophils; Cell Degranulation; Chemokine CXCL1; Chemokine CXCL2; Dermatitis, Contact; Ear; Eosinophils; Female; Filariasis; Filarioidea; Gerbillinae; Hypersensitivity, Immediate; Immune Complex Diseases; Immunoglobulin E; Immunoglobulin G; Immunosuppression Therapy; Inflammation; Leukocyte Count; Macrophages; Mast Cells; Mice; Mice, Inbred BALB C; Mice, Knockout; Neutrophils; Ovalbumin; Receptors, IgG

Dendritic cells (DCs) have the ability to present antigen and play a critical role in the induction of the acquired immune response. Skin DCs uptake antigen and subsequently migrate to regional draining lymph nodes (LNs), where they activate naive T cells. Here we show that the water/glycerol channel protein aquaporin 7 (AQP7) is expressed on epidermal and dermal DCs and involved in the initiation of primary immune responses. AQP7-deficient DCs showed a decreased cellular uptake of low-molecular-mass compounds (fluorescein isothiocyanate and Lucifer yellow) and high-molecular-mass substances (ovalbumin and dextran), suggesting that AQP7 is involved in antigen uptake. AQP7-deficient DCs also exhibited reduced chemokine-dependent cell migration in comparison to wild-type DCs. Consistent with these in vitro results, AQP7-deficient mice demonstrated a reduced accumulation of antigen-retaining DCs in the LNs after antigen application to the skin, which could be attributed to decreased antigen uptake and migration. Coincidentally, AQP7-deficient mice had impaired antigen-induced sensitization in a contact hypersensitivity model. These observations suggested that AQP7 in skin DCs is primarily involved in antigen uptake and in the subsequent migration of DCs and is responsible for antigen presentation and the promotion of downstream immune responses.

Topics: Animals; Antigens; Aquaporins; Cell Movement; Cells, Cultured; Chemotaxis; Dendritic Cells; Dermatitis, Contact; Dermis; Disease Models, Animal; Epidermal Cells; Epidermis; Glycerol; Haptens; Hypersensitivity; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Pinocytosis; Water

We previously reported that prior nasal administration of highly attenuated Bordetella pertussis BPZE1 provides effective and sustained protection against lethal challenge with influenza A viruses. The protective effect was mediated by suppressing the production of major pro-inflammatory mediators. To further explore the anti-inflammatory properties of BPZE1, we investigated the effect of BPZE1 nasal pretreatment on two mouse models of allergic disease, allergic airway inflammation, and contact hypersensitivity (CHS).. Allergic reactions were induced in mice nasally pretreated with live attenuated BPZE1 bacteria using the ovalbumin (OVA)-induced allergic airway inflammation and dinitrochlorobenzene (DNCB)-induced CHS models.. Prior BPZE1 nasal treatment suppressed OVA-induced lung inflammation and inflammatory cell recruitment and significantly reduced IgE levels and cytokine production. Similarly, BPZE1 nasal pretreatment markedly inhibited ear swelling, skin inflammation, and production of pro-inflammatory cytokines in the DNCB-induced CHS model. For both models, we showed that BPZE1 pretreatment does not affect the sensitization phase. Upon challenge, BPZE1 pretreatment selectively reduced the level of cytokines whose production is increased and did not affect the basal level of other cytokines. Together, our observations suggest that BPZE1 pretreatment specifically targets those cytokine-producing effector cells that are recruited and involved in the inflammatory reaction.. Our study demonstrates the broad anti-inflammatory properties of the attenuated B. pertussis BPZE1 vaccine candidate and supports its development as a promising agent to prevent and/or treat allergic diseases.

Topics: Administration, Intranasal; Animals; Bordetella pertussis; Cytokines; Dermatitis, Contact; Dinitrochlorobenzene; Disease Models, Animal; Female; Humans; Mice; Mice, Inbred BALB C; Ovalbumin; Pertussis Vaccine; Pneumonia; Vaccines, Attenuated; Whooping Cough

The interaction between CD27 and CD70 provides a costimulatory signal for T-cell survival. Although the role of CD27 signaling in CD8(+) T cells has been well defined, its role in CD4(+) T cells is relatively unknown. Here, we report that CD70 is specifically expressed on differentiated T-helper (Th)1 cells, but not on Th2 cells. Upon activation, CD70 expression increased markedly on Th1 cells, but remained undetectable on Th2 cells. We demonstrate that CD27 is involved in naive T-cell expansion in Th1-type, but not in Th2-type, immune responses as in vivo treatment with anti-CD70 monoclonal antibody at induction resulted in a significant reduction of delayed-type and contact hypersensitivity responses, but not asthmatic responses. In both Th1-type responses, during the priming phase, CD70 was detected at earlier time points on dendritic cells (DCs) and at later time points on CD4(+) T cells. Our results indicate that CD70 may be useful as a marker to distinguish Th1 from Th2 cells. More importantly, CD27 function may be controlled by the differentially regulated kinetics of CD70 expression on DCs and CD4(+) T cells, and Th1 cell-specific CD70 expression may be involved in an amplification loop for polarized Th1-type immune responses through T cell-T cell interactions.

Topics: Animals; Asthma; CD27 Ligand; Cytokines; Dermatitis, Contact; Female; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Ovalbumin; Th1 Cells; Th2 Cells; Tumor Necrosis Factor Receptor Superfamily, Member 7

New phenoxyacetic acid antagonists of CRTH2 are described. Following the discovery of a hit compound by a focused screening, high protein binding was identified as its main weakness. Optimization aimed at reducing serum protein binding led to the identification of several compounds that showed not only excellent affinities for the receptor (41 compounds with K(i) < 10 nM) but also excellent potencies in a human whole blood assay (IC(50) < 100 nM; PGD2-induced eosinophil shape change). Additional optimization of the PK characteristics led to the identification of several compounds suitable for in vivo testing. Of these, 19k and 19s were tested in two different pharmacological models (acute FITC-mediated contact hypersensitivity and ovalbumin-induced eosinophilia models) and found to be active after oral dosing (10 and 30 mg/kg).

Topics: Acetates; Administration, Oral; Alkynes; Animals; Anti-Allergic Agents; Anti-Inflammatory Agents; Binding, Competitive; Blood Proteins; Caco-2 Cells; Cell Membrane Permeability; Cell Shape; Chemotaxis, Leukocyte; Dermatitis, Contact; Eosinophils; Female; HEK293 Cells; Humans; Mice; Mice, Inbred BALB C; Microsomes, Liver; Ovalbumin; Phenoxyacetates; Protein Binding; Pulmonary Eosinophilia; Radioligand Assay; Rats; Receptors, Immunologic; Receptors, Prostaglandin; Structure-Activity Relationship; Sulfones

Tamoxifen (TX) represents the prototype selective oestrogen receptor modulator. In addition to its use in breast cancer, TX possesses immunomodulatory functions and displays beneficial effects in models of systemic lupus erythematosus. We hypothesized that TX might inhibit type I allergic reactions, which are also characterized by deviations in humoral immunity.. To evaluate the effects of TX on the allergic immune response in appropriate mouse models.. Balb/c mice were sensitized with ovalbumin (OVA)-alum by the intraperitoneal route, and humoral parameters, T cell cytokine patterns and OVA-induced ear swelling responses were determined in a preventive (start of TX treatment before sensitization) and a therapeutic setting (start after sensitization), respectively. In addition, the impact of TX on clinical signs, epidermal thickness and leucocyte infiltration of the skin was investigated in a model of allergen-induced dermatitis.. Preventive TX treatment interfered with all aspects of the allergic immune response, leading to a reduction of allergen-specific Ig levels (IgE, IgG1 and IgG2a), a skewing effect in the T cell compartment with the inhibition of IL-4 and an abrogation of ear swelling responses. Interestingly, a therapeutic TX administration was also effective in reducing Ig levels and ear swelling responses. The vigorous systemic effects were additionally mirrored by local changes in allergen-dependent dermatitis with reduced clinical symptoms, diminished epidermal thickness and decreased CD4+ and CD8+ cell infiltrates.. TX inhibits allergic responses when given preventively and also therapeutically, and improves allergen-induced dermatitis. Because of its effectiveness, TX could bear significant therapeutic potential for the treatment of allergies.

Topics: Allergens; Animals; Anti-Allergic Agents; Cytokines; Dermatitis, Contact; Female; Hypersensitivity; Immunoglobulin E; Mice; Mice, Inbred BALB C; Ovalbumin; T-Lymphocytes; Tamoxifen

Exposure to antigens of the fish parasite Anisakis is associated with the development of protein contact dermatitis in seafood-processing workers. Understanding the basic mechanisms controlling allergic sensitization through the skin is critical for designing therapies that will prevent the progression of allergic disease.. To investigate the roles of interleukin (IL)-4, IL-13 and the IL-4Ralpha in both local skin pathology and systemic sensitization following epicutaneous exposure to Anisakis proteins.. BALB/c wild-type (WT) mice and mice deficient in IL-4, IL-13 or IL-4 and IL-13, as well as mice with cell-specific impairment of IL-4Ralpha expression, were sensitized to Anisakis antigen by repeated epicutaneous application of Anisakis extract. Following this sensitization, skin pathology was recorded and systemic responses were investigated. Intravenous challenge with Anisakis extract was performed to test for the development of biologically relevant systemic sensitization.. In WT mice, epicutaneous sensitization with Anisakis larval antigens induced localized inflammation, epidermal hyperplasia, production of T(H)2 cytokines, antigen-specific IgE and IgG1. Intravenous challenge of sensitized mice resulted in anaphylactic shock. Interestingly, IL-13 deficient mice failed to develop epidermal hyperplasia and inflammation, whilst anaphylaxis was reduced only in strains deficient either in IL-4 only, or deficient in IL-4 and IL-13 concurrently, as well as in mice deficient in IL-4Ralpha or with impaired IL-4Ralpha expression on CD4(+) T cells.. Interleukin-13 plays a central role in protein contact dermatitis associated with repeated epicutaneous exposure to Anisakis extract, whereas IL-4 drives systemic sensitization and resultant anaphylactic shock.

Topics: Allergens; Anaphylaxis; Animals; Anisakis; Antibodies, Protozoan; Antigens, Helminth; CD4-Positive T-Lymphocytes; Dermatitis, Contact; Immunoglobulin E; Immunoglobulin G; Interleukin-13; Interleukin-4; Interleukin-4 Receptor alpha Subunit; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Skin

UV light can be highly beneficial in the treatment of skin disorders such as psoriasis. It is thought to cause immunosuppression by depleting or altering the function of epidermal Langerhans cells (LC). Our previous studies identified a novel langerin(+) dendritic cell in the dermis, distinct from LC in phenotype, circulation, and function. In this study, we determined the role of LC and dermal langerin(+) cells in UV suppression. UV light suppressed the CD8 T cell response to both contact hypersensitivity and epicutaneous protein immunization, and resulted in a dramatically altered phenotype of LC. UV light did not alter early CD8 T cell activation in the lymph nodes, but rather reduced CD8 T cell expansion at later time points. We found that dermal langerin(+) cells, but not LC, were essential for the CD8 T cell response. Furthermore, in the selective absence of LC, UV light still caused suppression of both CD8 T cell expansion and contact hypersensitivity.

Topics: Administration, Cutaneous; Animals; Antigens, Surface; CD8-Positive T-Lymphocytes; Dendritic Cells; Dermatitis, Contact; Epidermal Cells; Epidermis; Gene Knock-In Techniques; Immunosuppression Therapy; Langerhans Cells; Lectins, C-Type; Mannose-Binding Lectins; Mice; Mice, Inbred C57BL; Mice, Transgenic; Ovalbumin; Ultraviolet Rays

There are conflicting data in the literature regarding the role of epidermal Langerhans cells (LC) in promoting skin immune responses. On one hand, LC can be extremely potent APCs in vitro, and are thought to be involved in contact hypersensitivity (CHS). On the other hand, it seems counterintuitive that a cell type continually exposed to pathogens at the organism's barrier surfaces should readily trigger potent T cell responses. Indeed, LC depletion in one model led to enhanced contact hypersensitivity, suggesting they play a negative regulatory role. However, apparently similar LC depletion models did not show enhanced CHS, and in one case showed reduced CHS. In this study we found that acute depletion of mouse LC reduced CHS, but the timing of toxin administration was critical: toxin administration 3 days before priming did not impair CHS, whereas toxin administration 1 day before priming did. We also show that LC elimination reduced the T cell response to epicutaneous immunization with OVA protein Ag. However, this reduction was only observed when OVA was applied on the flank skin, and not on the ear. Additionally, peptide immunization was not blocked by depletion, regardless of the site. Finally we show that conditions which eliminate epidermal LC but spare other Langerin(+) DC do not impair the epicutaneous immunization response to OVA. Overall, our results reconcile previous conflicting data in the literature, and suggest that Langerin(+) cells do promote T cell responses to skin Ags, but only under defined conditions.

Topics: Animals; Antigens, Surface; Cell Proliferation; Dermatitis, Contact; Ear; Haptens; Kinetics; Lectins, C-Type; Mannose-Binding Lectins; Mice; Ovalbumin; Peptide Fragments; Ribs; Skin; T-Lymphocytes

The mechanisms by which erythemal UVB irradiation modulates systemic immune responses to antigens applied to non-irradiated sites are poorly understood. In this study, regulatory CD4+ T cells were identified in the skin-draining lymph nodes (SDLNs) of UVB-irradiated, but otherwise naive mice. A transgenic mouse strain (DO11.10) was utilized in which the majority of CD4+ T cells expressed the ovalbumin (OVA(323-339)) T-cell receptor. Thus, T-cell responses could be examined following erythemal UVB irradiation without further antigen sensitization. CD4+ T cells from the SDLNs of UVB-irradiated mice had significantly reduced capacity to respond to presentation of the OVA(323-339) peptide in vitro. Transfer of CD4+ T cells from the SDLNs of UVB-irradiated antigen-naive mice significantly reduced both OVA sensitization and contact hypersensitivity responses to an experimental hapten in the recipient mice. Depletion of CD4+CD25+ cells abrogated this UVB-suppressive effect in the in vitro proliferation assay. There was also a significant increase in the proportion of CD4+CD25+Foxp3+ cells in the SDLNs of UVB-irradiated mice. The potential of these regulatory cells poised to regulate responses to incoming antigens at distant non-irradiated sites broadens the biological impact of UVB irradiation of skin on immunity.

Topics: Animals; Antibody Formation; Antigens; CD4-Positive T-Lymphocytes; Cell Proliferation; Cells, Cultured; Dermatitis, Contact; Dose-Response Relationship, Radiation; Forkhead Transcription Factors; Immunization; Interleukin-2 Receptor alpha Subunit; Lymph Nodes; Mice; Mice, Inbred BALB C; Mice, Transgenic; Ovalbumin; Peptide Fragments; Receptors, Antigen, T-Cell; Time Factors; Ultraviolet Rays

Mast cells represent a potential source of TNF, a mediator which can enhance dendritic cell (DC) migration. Although the importance of mast cell-associated TNF in regulating DC migration in vivo is not clear, mast cells and mast cell-derived TNF can contribute to the expression of certain models of contact hypersensitivity (CHS). We found that CHS to FITC was significantly impaired in mast cell-deficient Kit(W-sh/W-sh) or TNF(-/)(-) mice. The reduced expression of CHS in Kit(W-sh/W-sh) mice was fully repaired by local transfer of wild-type bone marrow-derived cultured mast cells (BMCMCs), but was only partially repaired by transfer of TNF(-/)(-) BMCMCs. Thus, mast cells, and mast cell-derived TNF, were required for optimal expression of CHS to FITC. We found that the migration of FITC-bearing skin DCs into draining lymph nodes (LNs) 24 h after epicutaneous administration of FITC in naive mice was significantly reduced in mast cell-deficient or TNF(-/)(-) mice, but levels of DC migration in these mutant mice increased to greater than wild-type levels by 48 h after FITC sensitization. Mast cell-deficient or TNF(-/)(-) mice also exhibited significantly reduced migration of airway DCs to local LNs at 24 h after intranasal challenge with FITC-OVA. Migration of FITC-bearing DCs to LNs draining the skin or airways 24 h after sensitization was repaired in Kit(W-sh/W-sh) mice which had been engrafted with wild-type but not TNF(-/)(-) BMCMCs. Our findings indicate that mast cell-associated TNF can contribute significantly to the initial stages of FITC-induced migration of cutaneous or airway DCs.

Topics: Administration, Intranasal; Adrenal Cortex Hormones; Animals; Cell Movement; Cells, Cultured; Dendritic Cells; Dermatitis, Contact; Fluorescein-5-isothiocyanate; Lymph Nodes; Mast Cells; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Receptors, Tumor Necrosis Factor, Type I; Time Factors; Tumor Necrosis Factor-alpha

Coupling of ovalbumin (OVA) to anti-DEC-205 monoclonal antibody (mAb) (alphaDEC) induced the proliferation of OVA-specific T cells in vivo. Expansion was short-lived, caused by dendritic cells (DCs), and rendered T cells anergic thereafter. Phenotypic analysis revealed the induction of CD25+/CTLA-4+ T cells suppressing proliferation and interleukin-2 (IL-2) production of effector CD4+ T cells. The findings were supported by 2 disease models: (1) CD4+ T-cell-mediated hypersensitivity reactions were suppressed by the injection of alphaDEC-OVA and (2) the application of hapten-coupled alphaDEC-205 reduced CD8+ T-cell-mediated allergic reactions. Thus, targeting of antigens to immature DCs through alphaDEC antibodies led to the induction of regulatory T cells, providing the basis for novel strategies to induce regulatory T cells in vivo.

Topics: Animals; Antibodies, Monoclonal; Antigen Presentation; Antigens; Antigens, CD; Antigens, Differentiation; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; CTLA-4 Antigen; Dendritic Cells; Dermatitis, Contact; Haptens; Hypersensitivity; Hypersensitivity, Delayed; Interleukin-2; Lymphocyte Activation; Lymphocyte Count; Mice; Mice, Inbred BALB C; Ovalbumin; Receptors, Interleukin-2; T-Lymphocytes

Th2 responses are clearly involved in the pathogenesis of atopic disease. Thus, understanding the factors responsible for Th2 sensitization at sites of allergen exposure, such as airway and skin, is crucial for directing therapeutic or preventive strategies. Contrary to other models of Th2 sensitization to proteins, we have reported that Th2 responses induced by epicutaneous exposure to OVA are IL-4 independent. Combined deficiency of both IL-4 and IL-13 signaling did prevent Th2 generation, suggesting that IL-13 was mediating these IL-4-independent responses. It was not clear, however, whether IL-13 was simply replacing the need for IL-4 in genetically deficient mice or if IL-13 played a unique role. In the present study, we show that Th2 responses induced by epicutaneous OVA exposure (including lung inflammatory responses after inhaled Ag challenge, OVA-specific IgG1, and draining lymph node IL-5 production) are impaired in IL-13-deficient (IL-13(-/-)) mice compared with wild type. In contrast, i.p. sensitization of IL-13(-/-) mice resulted in responses equivalent to wild type. Generation of contact hypersensitivity to dinitrofluorobenzene, which involves Th1 and CD8(+) effector cells, was also intact in IL-13(-/-) mice. Taken together, the data indicate that IL-13 is the major inducer of Th2 generation in the cutaneous microenvironment, being required independently of IL-4. This fact, in combination with the known abundance of IL-13 in atopic dermatitis skin lesions, emphasizes the potentially important role of the skin as a site for Th2 sensitization to environmental allergens, particularly in atopic individuals.

Topics: Administration, Cutaneous; Administration, Intranasal; Animals; Antigens; Cytokines; Dermatitis, Contact; Dinitrofluorobenzene; Female; Haptens; Immunization; Immunoglobulin G; Inflammation; Injections, Intraperitoneal; Interleukin-13; Interleukin-4; Lung; Lymph Nodes; Mice; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; Skin; Solubility; STAT6 Transcription Factor; Th2 Cells; Trans-Activators

CX-659S ((S)-6-amino-5-(6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxamido)-3-methyl-1-phenyl-2,4(1H,3H)-pyrimidinedione), a newly discovered anti-inflammatory compound, exerts inhibitory effects against picryl chloride-, oxazolone-, and dinitrochlorobenzene-induced acute contact hypersensitivity responses (CHRs) characterized by Th1-type reactions. Furthermore, this compound suppressed chronic CHRs characterized by Th2-type reactions, which is well known to mimic many, if not all, events occurring within the lesional skin of patients with atopic dermatitis (AD). The present study was conducted to determine the combined effect of topical CX-659S with topical corticosteroid on immediate type (ITR), late type (LTR), and delayed type hypersensitivity (DTHR) allergic reactions that are involved in AD. An ineffective dose of CX-659S (0.03 mg/ear) combined with betamethasone valerate (BV) significantly potentiated inhibitory activity of BV alone (0.1 micro g/ear and 0.3Shizuokag/ear) on both the ITR and the LTR in mice with the ovalbumin (OVA)-induced biphasic cutaneous reaction. Furthermore, the combined effect of CX-659S with BV was also observed on dinitrochlorobenzene (DNCB)-induced DTHR in guinea pigs. These results indicate that CX-659S has a combined effect with corticosteroids on every ITR, LTR, and DTHR. Proper treatment with corticosteroids for a safe and effective treatment of AD is needed. Thus, the combination therapy of topical CX-659S with topical corticosteroid would be one of the potential approaches for devising a proper treatment with corticosteroids.

Topics: Administration, Topical; Animals; Anti-Inflammatory Agents; Anti-Inflammatory Agents, Non-Steroidal; Betamethasone Valerate; Dermatitis, Contact; Dinitrochlorobenzene; Guinea Pigs; Hypersensitivity, Delayed; Hypersensitivity, Immediate; Indicators and Reagents; Irritants; Mice; Mice, Inbred ICR; Ovalbumin; Skin; Uracil

The migration of dendritic cells (DCs) from the epithelia to the lymphoid organs represents a tightly regulated multistep event involved in the induction of the immune response. In this process fatty acid derivatives positively and negatively regulate DC emigration. In the present study we investigated whether activation of peroxisome proliferator-activated receptors (PPARs), a family of nuclear receptors activated by naturally occurring derivatives of arachidonic acid, could control DC migration from the peripheral sites of Ag capture to the draining lymph nodes (DLNs). First, we show that murine epidermal Langerhans cells (LCs) express PPAR gamma, but not PPAR alpha, mRNA, and protein. Using an experimental murine model of LC migration induced by TNF-alpha, we show that the highly potent PPAR gamma agonist rosiglitazone specifically impairs the departure of LCs from the epidermis. In a model of contact allergen-induced LC migration, PPAR gamma activation not only impedes LC emigration, and their subsequent accumulation as DCs in the DLNs, but also dramatically prevents the contact hypersensitivity responses after challenge. Finally, after intratracheal sensitization with an FITC-conjugated Ag, PPAR gamma activation inhibits the migration of DCs from the airway mucosa to the thoracic LNs and also profoundly reduces the priming of Ag-specific T lymphocytes in the DLNs. Our results suggest a novel regulatory pathway via PPAR gamma for DC migration from epithelia that could contribute to the initiation of immune responses.

Topics: Animals; Cell Migration Inhibition; Cell Movement; Cells, Cultured; Cytokines; Dendritic Cells; Dermatitis, Contact; Epidermis; Female; Fluorescein-5-isothiocyanate; Haptens; Immunosuppressive Agents; Langerhans Cells; Lung; Lymph Nodes; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Mice, Transgenic; Ovalbumin; Receptors, Cytoplasmic and Nuclear; Rosiglitazone; Thiazoles; Thiazolidinediones; Thorax; Transcription Factors; Transcriptional Activation; Tumor Necrosis Factor-alpha

IFN-gamma-inducible protein 10 (IP-10, CXCL10), a chemokine secreted from cells stimulated with type I and II IFNs and LPS, is a chemoattractant for activated T cells. Expression of IP-10 is seen in many Th1-type inflammatory diseases, where it is thought to play an important role in recruiting activated T cells into sites of tissue inflammation. To determine the in vivo function of IP-10, we constructed an IP-10-deficient mouse (IP-10(-/-)) by targeted gene disruption. Immunological analysis revealed that IP-10(-/-) mice had impaired T cell responses. T cell proliferation to allogeneic and antigenic stimulation and IFN-gamma secretion in response to antigenic challenge were impaired in IP-10(-/-) mice. In addition, IP-10(-/-) mice exhibited an impaired contact hypersensitivity response, characterized by decreased ear swelling and reduced inflammatory cell infiltrates. T cells recovered from draining lymph nodes also had a decreased proliferative response to Ag restimulation. Furthermore, IP-10(-/-) mice infected with a neurotropic mouse hepatitis virus had an impaired ability to control viral replication in the brain. This was associated with decreased recruitment of CD4(+) and CD8(+) lymphocytes into the brain, reduced levels of IFN-gamma and the IFN-gamma-induced chemokines monokine induced by IFN-gamma (Mig, CXCL9) and IFN-inducible T cell alpha chemoattractant (I-TAC, CXCL11) in the brain, decreased numbers of virus-specific IFN-gamma-secreting CD8(+) cells in the spleen, and reduced levels of demyelination in the CNS. Taken together, our data suggest a role for IP-10 in both effector T cell generation and trafficking in vivo.

Topics: Animals; Antigens; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Movement; Chemokine CXCL10; Chemokines, CXC; Coronavirus Infections; Demyelinating Diseases; Dermatitis, Contact; Down-Regulation; Encephalomyelitis; Growth Inhibitors; Interferon-gamma; Isoantigens; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Lymphocyte Depletion; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Murine hepatitis virus; Mutagenesis, Site-Directed; Ovalbumin; Spleen

The paucity of lymph node T cells (plt) mutation leads to a loss of CCL21 and CCL19 expression in secondary lymphoid organs. plt mice have defects in the migration of naive T cells and activated dendritic cells into the T cell zones of lymphoid organs, suggesting that they would have defects in T cell immune responses. We now demonstrate T cell responses in plt mice are delayed but ultimately enhanced. Responses to contact sensitization are decreased at day 2 after priming but increased at day 6. After subcutaneous immunization, antigen-specific T cell proliferation and cytokine production in plt mice are increased and remain markedly elevated for at least 8 wk. Compared with wild-type mice, a proportion of T cell response in plt mice are shifted to the spleen, and prior splenectomy reduces the T cell response in draining lymph nodes. After immunization of plt mice, T cells and dendritic cells colocalize in the superficial cortex of lymph nodes and in splenic bridging channels, but not in T cell zones. These results demonstrate that plt mice mount robust T cell responses despite the failure of naive T cells and activated dendritic cells to enter the thymus dependent areas of secondary lymphoid organs.

Topics: Animals; Chemokine CCL19; Chemokine CCL21; Chemokines, CC; Dendritic Cells; Dermatitis, Contact; Immunity, Cellular; Immunization; In Vitro Techniques; Lymphocyte Activation; Lymphoid Tissue; Mice; Mice, Inbred BALB C; Mice, Mutant Strains; Mutation; Ovalbumin; T-Lymphocytes; Time Factors

Standard protocols to generate mouse dendritic cells (DC) generally use culture medium supplemented with fetal calf serum; however, reinjection in vivo of DC cultured in fetal calf serum results in priming to xenogeneic proteins that clearly limits the use of such DC. We therefore established a fetal calf serum-free culture system for the generation of murine DC from bone marrow precursors. DC can be generated fetal calf serum-free using RPMI supplemented with 1.5% syngeneic mouse serum. Although the yield of DC grown under fetal calf serum-free conditions was somewhat lower than that of the standard culture, large numbers of DC could be generated without the exposure to xenogeneic proteins. The yield of fetal calf serum-free cultured DC was further enhanced by addition of the proinflammatory cytokines TNF-alpha and IL-1beta with the combination resulting in up to 10% more DC. Phenotypically, CD11c + DC cultured fetal calf serum-free homogenously coexpressed the DC-specific molecule DEC-205 as well as the costimulatory molecules CD40, CD80, and CD86. In contrast, only a subpopulation of the CD11c + DC cultured in fetal calf serum-containing medium coexpressed these molecules. Functionally, fetal calf serum-free DC showed strong stimulatory capacity for naïve allogeneic CD4 + and CD8 + T cells. Importantly, fetal calf serum-free DC showed spontaneous in vivo migratory activity. Moreover, 5 x 105 subcutaneously injected TNBS-conjugated fetal calf serum-free DC were able to mediate contact sensitivity. Furthermore, the intravenous or subcutaneous injection of a single dose of 5 x 105 OVA-pulsed fetal calf serum-free DC resulted in the induction of an OVA-specific immune response in naïve TCR transgenic animals. Thus DC cultured under fetal calf serum-free conditions are suitable instruments for in vivo therapeutic approaches, especially in autoimmune models.. DC vaccines/dendritic cell development/fetal calf serum-free culture conditions for DC/in vivo therapeutic DC approaches.

Topics: Animals; Bone Marrow Cells; Cattle; Cell Count; Cell Division; Cell Movement; Culture Media; Dendritic Cells; Dermatitis, Contact; Fetal Blood; Interleukin-1; Mice; Mice, Inbred Strains; Mice, Transgenic; Ovalbumin; Phenotype; Receptors, Antigen, T-Cell; Stem Cells; Tumor Necrosis Factor-alpha

1. Small, N- to C-terminal cyclized peptides containing the leucyl-aspartyl-valine (LDV) motif from fibronectin connecting segment-1 (CS-1) have been investigated for their effects on the adhesion of human T-lymphoblastic leukaemia cells (MOLT-4) to human plasma fibronectin in vitro mediated by the integrin Very Late Antigen (VLA)-4 (alpha4beta1, CD49d/CD29). 2. Cyclo(-isoleucyl-leucyl-aspartyl-valyl-aminohexanoyl-) (c(ILDV-NH(CH2)5CO)) was approximately 5 fold more potent (IC50 3.6+/-0.44 microM) than the 25-amino acid linear CS-1 peptide. Cyclic peptides containing two more or one less methylene groups had similar potency to c(ILDV-NH(CH2)5CO) while a compound containing three less methylene groups, c(ILDV-NH(CH2)2CO), was inactive at 100 microM. 3. c(ILDV-NH(CH2)5CO) had little effect on cell adhesion mediated by two other integrins, VLA-5 (alpha5,beta1, CD49e/CD29) (K562 cell adhesion to fibronectin) or Leukocyte Function Associated molecule-1 (LFA-1, alphabeta2, CD11a/CD18) (U937 cell adhesion to Chinese hamster ovary cells transfected with intercellular adhesion molecule-1) at concentrations up to 300 microM. 4. c(ILDV-NH(CH2)5CO) inhibited ovalbumin delayed-type hypersensitivity or oxazolone contact hypersensitivity in Balb/c mice when dosed continuously from subcutaneous osmotic mini-pumps (0.1-10 mg kg(-1) day(-1)). Maximum inhibition (approximately 40%) was similar to that caused by the monoclonal antibody PS/2 (7.5 mg kg(-1) i.v.) directed against the alpha4 integrin subunit. 5. c(ILDV-NH(CH2)5CO) also inhibited oxazolone contact hypersensitivity when dosed intravenously 20 h after oxazolone challenge (1-10 mg kg(-1)). Ear swelling was reduced at 3 h and 4 h but not at 1 h and 2 h post-dose (10 mg kg(-1)). 6. Small molecule VLA-4 inhibitors derived from c(ILDV-NH(CH2)5CO) may be useful as anti-inflammatory agents.

Topics: Amino Acid Sequence; Animals; Anti-Inflammatory Agents, Non-Steroidal; Cell Adhesion; CHO Cells; Cricetinae; Dermatitis, Contact; Female; Fibronectins; Humans; Hypersensitivity, Delayed; Inflammation; Integrin alpha4beta1; Integrins; Intercellular Adhesion Molecule-1; Intercellular Signaling Peptides and Proteins; Leukemia, Erythroblastic, Acute; Leukemia, T-Cell; Mice; Mice, Inbred BALB C; Molecular Sequence Data; Ovalbumin; Oxazolone; Peptides; Rats; Receptors, Lymphocyte Homing; T-Lymphocytes; Transfection

Exposure to ultraviolet light, especially UVB wavelengths, can impair immune responses in animals and humans. It is remarkable that this immunomodulation is not restricted to the exposed skin but is also found at other sites, i.e. systemic (distant) immunosuppression. A frequently proposed hypothesis is that UVB exposure inhibits, specifically, T helper 1 (Th1)-mediated immune responses. The major reason for this is that contact hypersensitivity (CHS) and delayed-type hypersensitivity (DTH), both Th1-mediated immune responses, are very sensitive to UVB. For this reason these models are frequently used for photoimmunology studies. In the present study, the effects of UVB exposure were investigated in classical models for Th1-mediated immunity, i.e. CHS models in which picrylchloride or oxazolone were used as low-molecular-weight chemical antigens. In these models, CHS responsiveness and cytokines were measured, the latter by both reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). The CHS responses to both contact sensitizers (picrylchloride and oxazolone) were suppressed significantly by pre-exposure to repeated suberythemal UVB exposure. Interferon-gamma (IFN-gamma), interleukin (IL)-12 and IL-4, but not IL-10, were detectable in spleen and draining lymph nodes of sensitized BALB/c mice. Repeated UVB exposure prior to sensitization at a distant locus inhibited both IFN-gamma and IL-12 but not IL-4. In BALB/c mice sensitized with ovalbumin (OVA) in the absence of complete Freund's adjuvant, a model for Th2-mediated immunity, OVA-specific serum IgE and cytokine profiles in the spleen were analysed. Sensitization did lead to a significant increase in OVA-specific IgE serum titres. Pre-exposure to UVB resulted in a decreased OVA-specific IgE serum titre. Both RT-PCR and ELISA showed increased levels of IFN-gamma, IL-4 and IL-10 in the spleens of OVA-sensitized mice. The production of IFN-gamma and IL-4 was not affected by UVB pre-exposure. In contrast, the production of IL-10 was significantly increased. This was probably caused by an up-regulation of Th2 cells. It is remarkable that IFN-gamma is significantly suppressed by UVB in Th1-mediated immune reactions but not in Th2-mediated immune reactions where it even appears to increase. IL-10, which is up-regulated by UVB pre-exposure and produced by, among others, Th2 cells, may represent a shift from Th1- to Th2-mediated immune mechanisms. However, IL-10 ca

Topics: Animals; Cytokines; Dermatitis, Contact; Enzyme-Linked Immunosorbent Assay; Interferon-gamma; Interleukin-4; Lymph Nodes; Male; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Reverse Transcriptase Polymerase Chain Reaction; Spleen; Th1 Cells; Th2 Cells; Ultraviolet Rays

The tumor necrosis factor family molecule Ox40-ligand (Ox40L) has been identified as a potential costimulatory molecule and also has been implicated in T cell homing and B cell activation. To ascertain the essential functions of Ox40L, we generated and characterized Ox40L-deficient mice. Mice lacking Ox40L exhibit an impaired contact hypersensitivity response, a dendritic cell-dependent T cell-mediated response, due to defects in T cell priming and cytokine production. In contrast, Ox40L-deficient mice do not have defects in T cell homing or humoral immune responses. In vitro, Ox40L-deficient dendritic cells are defective in costimulating T cell cytokine production. Thus, Ox40L has a critical costimulatory function in vitro and in vivo for dendritic cell:T cell interactions.

Topics: 3T3 Cells; Animals; Antigens, T-Independent; Dendritic Cells; Dermatitis, Contact; Haptens; Hemocyanins; Hypersensitivity, Delayed; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Knockout; Ovalbumin; OX40 Ligand; Receptors, Tumor Necrosis Factor; T-Lymphocytes; Tumor Necrosis Factors

Langerhans cells (LCs) are antigen-presenting cells of the skin that trap small contact-sensitizing molecules and induce cutaneous hypersensitivity. LCs can present larger molecules but the mechanisms of processing have required investigation. A system combining in vitro culture of antigen with epidermal cells in the presence of inhibitors, followed by fixation and transfer of these antigen/drug-treated epidermal cells to naive mice, was developed to investigate the steps of antigen processing. Langerhans cells undertake similar, but not identical, pathways for the processing of simple and complex molecules. Complex molecules such as trinitrophenyl conjugated to ovalbumin (TNP-OVA) were internalized and modification required a chloroquine-sensitive proteolysis step and a cycloheximide-sensitive protein synthesis step. This modified product was actively recycled to the cell membrane as presentation was inhibited by blocking receptor translocation with either monensin or cytochalasin B. Small contact sensitizers such as trinitrophenyl did not undergo modification but required internalization and presentation was also inhibited by blocking receptor translocation.

Topics: Animals; Antigen Presentation; Chloroquine; Cycloheximide; Cytochalasin B; Dermatitis, Contact; Female; Langerhans Cells; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Monensin; Ovalbumin; Skin; Trinitrobenzenes

Delayed hypersensitivity (DH) is an important immune effector modality that successfully wards off intracellular pathogens and many parasites, but also causes immunopathogenic injury to vital tissues. Particularly in the eye, DH has devastating effects that can lead to blindness. Ags injected into the anterior chamber of the eye of naive mice elicit a deviant form of systemic immunity in which DH is selectively down-regulated. Expression of DH in this model system is curtailed by regulatory CD8+ T cells. At present, we have determined whether injection of Ag into the anterior chamber of eyes of specifically sensitized mice also impairs DH expression. Our results indicate that DH is blunted or eliminated in previously primed mice when heterologous proteins, retinal autoantigens, or minor histocompatibility Ags are injected into the anterior chamber. Suppression is achieved in this system by Ag-specific CD8+ T cells, and failed DH can be imposed on immunized mice by i.v. injections of peritoneal exudate cells pulsed with Ag in vitro in the presence of TGF-beta. Thus, the immune regulatory mechanisms that operate to protect the eye from immunogenic inflammation can be invoked in previously sensitized mice. In addition, tolerance could not be generated in presensitized mice by either i.v. injection of soluble Ag or painting of hapten on UVB-exposed skin. It seems that the strategies used by the eye to create a deviant state of immunity in the face of pre-existing conventional immunity may be unique.

Topics: Animals; Anterior Chamber; Dermatitis, Contact; Eye Proteins; Hypersensitivity, Delayed; Immune Tolerance; Mice; Mice, Inbred BALB C; Neoplasm Transplantation; Ovalbumin; Retinol-Binding Proteins; Skin; Solubility; T-Lymphocytes, Regulatory; Transforming Growth Factor beta; Ultraviolet Rays

To confirm positivity in routine guinea pig studies, contact allergenicity was investigated by a challenge assay in vitro using a co-culture of autologous lymphocytes passed through a nylon-wool column and antigen-presenting cells (APCs) modified with or without antigen. Proliferation of the lymphocytes primed with ovalbumin and/or 2,4-dinitrochlorobenzene was antigen specific and dependent on the presence of APCs (peripheral blood monocytes, splenic macrophages and macrophages induced by liquid paraffin). For another nine haptens, primed lymphocytes proliferated significantly more than control lymphocytes; the stimulation index (SI; ratio between [3H]methylthymidine ([3H]TdR) incorporation of lymphocytes with antigen-modified APCs and [3H]TdR incorporation of lymphocytes with APCs not modified by antigen) was 1.6-4.8 in sensitized animals whereas it was about 1.0 in control animals. Sodium dodecyl sulfate did not cause lymphocyte proliferation. The SI value in vitro was correlated with both the positive rate in vivo (r = 0.736) and the mean response score in vivo (r = 0.645). Thus, it was possible to confirm that positivity in routine experiments was a true sign of allergy. A combination of this assay and short-term animal studies would provide an efficient assessment of the allergic potential of chemicals.

Topics: Animals; Antigen-Presenting Cells; Cell Division; Dermatitis, Contact; Female; Guinea Pigs; Haptens; Immunologic Tests; In Vitro Techniques; Lymphocyte Activation; Macrophages; Mineral Oil; Monocytes; Ovalbumin; Spleen

We previously showed that BALB/c mice sensitized to ovalbumin (OVA) by brief daily inhalations of antigen over 10 consecutive days exhibit elevated antigen-specific serum IgE antibody levels and increased airways responsiveness. For the first time, we now show that animals sensitized in this fashion to either OVA or ragweed (RGW) develop immediate hypersensitivity skin test reactions when challenged 2 d after completion of the sensitization protocol. Skin testing, performed by direct assessment of wheal formation after intradermal injection of allergen, was sensitive and specific, since animals exposed to RGW by inhalation only responded to RGW, and OVA-sensitized animals responded only to OVA. Positive reactions were associated with mast cell degranulation, whereas control injections were not. Since only sensitized IgE high responder BALB/c mice but neither nonsensitized BALB/c mice nor OVA-sensitized IgE low responder SJL/J mice exhibited wheal responses, induction of OVA-specific IgE appeared to be essential for the mediation of OVA-specific immediate hypersensitivity reactions of the skin in this model. Passive cutaneous anaphylaxis (PCA) testing confirmed the presence of antigen-specific IgE in the serum. Mice that developed IgG (predominantly IgG2b) anti-OVA antibodies did not respond to OVA injection, indicating that OVA-specific IgG was not involved in this system. Further support for the role of IgE in the immediate hypersensitivity response included the wheal response to intradermal injection of anti-IgE antibody that occurred in OVA- and RGW-sensitized mice at 10-fold lower concentrations than in nonsensitized BALB/c mice and not in sensitized SJL/J mice. After transfer of mononuclear cells from peribronchial lymph nodes of OVA- or RGW-sensitized BALB/c mice, naive, syngeneic recipients developed antigen-specific IgE and specific immediate hypersensitivity responses, indicating that the local lymphoid tissue at the site of sensitization can transfer responsiveness to these allergens. These results demonstrate for the first time the ability to elicit and study IgE-mediated immediate skin hypersensitivity responses in the mouse and illustrate the association of increased antigen-specific and total serum IgE levels, airways hyperresponsiveness, and antigen-specific immediate cutaneous reactivity after sensitization to allergen via the airways.

Topics: Aerosols; Animals; Antigens; Dermatitis, Contact; Female; Hypersensitivity, Immediate; Immunization; Immunoglobulin E; Immunotherapy, Adoptive; Mast Cells; Mice; Mice, Inbred BALB C; Mice, Inbred Strains; Ovalbumin; Skin; Skin Tests; Time Factors

An animal study was conducted to elucidate the role of ovalbumin (OA) in the development of eczematous lesions in intrauterine sensitized newborns. Four groups of pregnant guinea pigs were used: group A, immunized by oral administration of 1% OA in drinking water until parturition; group B, immunized by intradermal injection of OA with Freund's complete adjuvant; group C, immunized by both methods; and group D (control), not immunized. The newborn guinea pigs of each group were patch tested with 10% OA in white petrolatum. Positive reactions were seen in the newborns of groups B and C, but not in those in groups A and D. By enzyme-linked immunosorbent assay and passive cutaneous anaphylaxis, a high titre of OA-specific IgG was detected in the group B and C newborns. The number of positive patch test reactions decreased concomitantly with the decline of specific IgG. Histologically, eczematous changes were observed in the positive reaction sites. Many OA antigen-bearing Langerhans cells were found by the immuno-double labelling technique. Immuno-electron microscopic findings revealed the presence of OA antigens as well as IgG molecules on the cytoplasmic membranes of Langerhans cells. Our studies demonstrated that maternal sensitization with OA can induce an eczematous reaction in the newborns to OA patch testing under the presence of high levels of OA-specific IgG in the serum. From these findings it is suggested that IgG plays an essential role in the development of contact hypersensitivity reaction to OA.

Topics: Administration, Oral; Animals; Animals, Newborn; Dermatitis, Contact; Disease Models, Animal; Female; Food Hypersensitivity; Guinea Pigs; Immunity, Maternally-Acquired; Immunoglobulin G; Injections, Intradermal; Male; Ovalbumin; Passive Cutaneous Anaphylaxis; Patch Tests; Pregnancy

Offspring of mice treated with cyclophosphamide (Cy; 1, 2.5 or 5 mg/kg) during pregnancy (6-18 days of gestation) and tested for immunocompetence from 5 to 10 weeks of age were found to have defective reticuloendothelial clearance. The main effects were: a) increased elimination half time (T 1/2) of 51Cr-labeled SRBC from circulation, b) decreased liver uptake of 51Cr and c) impaired ability of the spleen, mostly affecting the female pups, to compensate for decreased liver uptake. The highest dose group suffered the most pronounced effects. This group was also found to have increased IgG immunoglobulin levels at 7 weeks of age. IgG antibody production in response to specific antigenic stimulation and delayed hypersensitivity reactions to oxazolone did not appear to be affected by Cy treatment.

Topics: Aging; Animals; Body Weight; Chromium Radioisotopes; Cyclophosphamide; Dermatitis, Contact; Female; Immunocompetence; Immunoglobulin G; Immunoglobulin M; Mice; Mononuclear Phagocyte System; Ovalbumin; Oxazolone; Pregnancy; Prenatal Exposure Delayed Effects; Reticulocytes; Sheep

Basophils are the main cell component in cutaneous basophil hypersensitivity (CBH) reactions, but the role of basophils and the factors which gather them into CBH reaction sites are unknown. To investigate these problems, we induced CBH reactions in guinea pigs and observed basophils and mast cells in the skin reaction sites using light and electron microscopy. Basophils infiltrated into CBH reaction site appeared at 5 h after the challenge with antigen, increased till 48 h and decreased thereafter. On the other hand, the number of mast cells and their granules decreased after the challenge with antigen and reached a minimum at 48 h, but recovered at 96 h. The changes in the number of basophils and mast cells were complementary. This result suggested that the granules of mast cells may have a factor to gather basophils into the CBH skin reaction site. Furthermore, basophils infiltrated in the CBH reaction site were degranulated by the rechallenge with antigen, which was considered to be by an anaphylactic reaction.

Topics: Animals; Basophils; Cell Movement; Dermatitis, Contact; Female; Guinea Pigs; Leukocyte Count; Mast Cells; Ovalbumin; Skin

In previous studies, the alpha 2-adrenoceptor agonist clonidine has been shown to suppress the wheal and flare reaction in guinea pigs sensitized to ovalbumin. This phenomenon has been further studied with special reference to effects on the dermal inflammatory cell infiltrate and mast cells. Clonidine lessens the degranulation of mast cells seen in control untreated immediate hypersensitivity reactions. Less neutrophils and eosinophils arrive to the treated reactions. Basophils and mononuclear cells (chiefly lymphocytes) which characterize the late phase of the wheal and flare reaction were not influenced by clonidine. Clonidine had a possible minimal effect on allergic contact (delayed hypersensitivity) reactions. The toxic contact reaction to croton oil (nonspecific cutaneous inflammation) was not affected.

Topics: Animals; Cell Movement; Clonidine; Croton Oil; Dermatitis, Contact; Drug Hypersensitivity; Female; Granulocytes; Guinea Pigs; Histamine; Hypersensitivity, Delayed; Hypersensitivity, Immediate; Inflammation; Mast Cells; Ovalbumin; Oxazolone; Skin; Skin Tests

Administration of cyclosporin A (CsA; 25 mg/kg) orally to guinea pigs from the time of immunization with ovalbumin (OVA) in complete Freund's adjuvant, followed by drug withdrawal 4 days later, resulted in marked potentiation of classical, tuberculin-like delayed-hypersensitivity skin responses to OVA. However, no such augmentation of delayed-type hypersensitivity (DTH) to purified protein derivative (PPD) was demonstrated. The enhancing effect of CsA was also dependent on the dose of OVA used for both immunization and skin testing and on the interval between drug withdrawal and the elicitation of DTH. A single intraperitoneal injection of CsA (200 mg/kg) given 2 days before immunization also had an augmentary effect on 14-day responses to OVA. Similar treatment protocols, however, did not enhance Jones-Mote (cutaneous basophil) hypersensitivity to OVA or contact sensitivity reactions to dinitrofluorobenzene. Longer courses of CsA (25 mg/kg per os) between sensitization and skin testing severely depressed all three categories of type IV hypersensitivity reactions. Our observations may have important cautionary implications for the prospective management of immunologically mediated diseases of intermittent activity, including certain autoimmune disorders, where short courses of CsA might be contemplated.

Topics: Animals; Cyclosporins; Dermatitis, Contact; Female; Guinea Pigs; Hypersensitivity, Delayed; Immunization; Ovalbumin; Skin; Skin Tests; Tuberculin

Topics: Animals; Cyclophosphamide; Cyclosporins; Dermatitis, Contact; Dinitrofluorobenzene; Drug Hypersensitivity; Guinea Pigs; Hypersensitivity, Delayed; Immunity, Cellular; Immunosuppression Therapy; Male; Ovalbumin; Skin Tests; T-Lymphocytes, Regulatory

An in vitro histamine release assay was used to test the hypothesis that passive sensitization of circulating basophils is associated with the activity of immune serum that transfer the ability to elicit cutaneous basophil hypersensitivity (CBH) reactions. Systemic i.v. transfer of several types of immune sera that mediate CBH also led to passive sensitization of circulating basophils for antigen-specific release of histamine in vitro. In addition, we found that immune serum passively sensitizes basophils in vitro. Thus immune sera had three activities that are probably interconnected: sera will 1) passively transfer CBH in vivo, 2) passively sensitize basophils in vivo, and 3) passively sensitize basophils in vitro. These results suggest that passive sensitization of circulating basophils by immune serum contributes to the mechanism by which antibodies transfer the ability to elicit CBH reactions.

Topics: Animals; Basophils; Dermatitis, Contact; Epitopes; Female; Guinea Pigs; Hemocyanins; Histamine Release; Immune Sera; Immunization; Immunization, Passive; Injections, Intradermal; Ovalbumin

A new attenuation marker to distinguish a virulent strain (XJJV) from an attenuated strain (XJC13JV or XJOJV) of Junin virus by means of the humoral and cellular responses to unrelated antigens was studied in guinea pigs. Strain XJJV suppressed the humoral immune response, as shown by the lower titers of precipitating antibody to ovalbumin. The concomitant decrease in serum complement level contributed to a milder Arthus cutaneous reactivity. In contrast, the attenuated strains did not decrease the humoral response. The pathogenic strain suppressed cell-mediated immunity, as demonstrated by decreased contact sensitivity to 2,4-dinitro-1-fluorobenzene and by depression of delayed skin reactions to tuberculin purified protein derivative. When attenuated strains were used, such suppressive effects were not observed. For virulent strain XJJV, virus replication in lymphoid organs and immunosuppressive effects were correlated. These findings provide a further means to differentiate between virulent and attenuated strains of Junin virus for the purpose of vaccine control of Argentine hemorrhagic fever.

Topics: Animals; Antibodies; Antibody Formation; Arenaviridae; Arenaviruses, New World; Arthus Reaction; Dermatitis, Contact; Dinitrofluorobenzene; Female; Guinea Pigs; Immunity, Cellular; Male; Ovalbumin; Tuberculin Test; Vaccines, Attenuated; Virulence

DNA--guinea-pig erythrocyte rosette-forming cells (RFC), as found after contact sensitization of guinea-pigs with DNCB, were shown to be highly sensitive to pretreatment with cyclophosphamide or X-irradiation, which suggests that these cells are B cells. The finding was confirmed by rosette-blocking experiments using an anti-immunoglobulin serum. It has already been shown that they are not directly related to antibody production. Possibly they represent precursors of antibody-producing cells. An increase in rosette formation, using non-antigenically related erythrocytes, was shown to parallel development of cell-mediated immunity (CMI) in the lymph nodes. These non-specifically induced RFC were also found to be B cells. Data were obtained which make it improbable that either proliferation or influx of B cells during development of CMI plays a major role in this increase. An alternative mechanism is suggested, while a possible role for these non-specifically induced RC is proposed.

Topics: Animals; Antibody Formation; Antibody-Producing Cells; Antilymphocyte Serum; B-Lymphocytes; Cyclophosphamide; Dermatitis, Contact; Dinitrochlorobenzene; Erythrocytes; Female; Guinea Pigs; Hemolytic Plaque Technique; Immune Adherence Reaction; Immunity, Cellular; Immunosuppression Therapy; Lymph Nodes; Nitrobenzenes; Ovalbumin; Oxazolone; Polysaccharides, Bacterial; Spleen; T-Lymphocytes

Topics: Administration, Topical; Amylases; Animals; Antibodies, Anti-Idiotypic; Antibody Formation; Antigens; Azo Compounds; Bacillus subtilis; Blood Proteins; Carrier Proteins; Cyclic N-Oxides; Dermatitis, Contact; Erythrocytes; Female; Haptens; Hemagglutination Tests; Hydroxylamines; Immunity, Cellular; Immunization, Passive; Immunosuppression Therapy; Injections, Subcutaneous; Mice; Ovalbumin; Picryl Chloride; Quinolines; Rabbits

Topics: Animals; B-Lymphocytes; Cyclophosphamide; Dermatitis, Contact; Fluorine; Guinea Pigs; Hypersensitivity, Delayed; Immunity, Cellular; Immunosuppression Therapy; Nitrobenzenes; Ovalbumin; Skin Tests; Spleen; T-Lymphocytes; Time Factors

Topics: Animals; Antibody Formation; Antigens; Antilymphocyte Serum; B-Lymphocytes; Cyclophosphamide; Dermatitis, Contact; Guinea Pigs; Hypersensitivity, Delayed; Immunity, Cellular; Lymph Nodes; Mice; Mitosis; Nitrophenols; Ovalbumin; Skin Tests; Spleen; Splenectomy; T-Lymphocytes; Tuberculin Test

Topics: Animals; Antibody Formation; Antibody Specificity; Antigen-Antibody Reactions; Antigens; Carbon Isotopes; Chemical Phenomena; Chemistry; Chromatography, Ion Exchange; Dermatitis, Contact; Drug Hypersensitivity; Formaldehyde; Goats; Haptens; Hypersensitivity, Delayed; Immunodiffusion; Immunoelectrophoresis; Muramidase; Ovalbumin; Rabbits; Serum Albumin

Topics: Aniline Compounds; Animals; Ascitic Fluid; Dermatitis, Contact; Ear, External; Freund's Adjuvant; gamma-Globulins; Hypersensitivity, Delayed; Immune Tolerance; Immunity, Maternally-Acquired; Immunization; Lymph Nodes; Mice; Mycobacterium bovis; Mycobacterium tuberculosis; Nitrobenzenes; Ovalbumin; Oxazoles; Picryl Chloride; Sulfonic Acids; Tuberculin

In earlier observations with the picryl system, it was concluded that contact sensitivity was a form of delayed (cellular) hypersensitivity to conjugates of sensitizer with autologous proteins indistinguishable in its immunological mechanism from other classical forms of delayed hypersensitivity to proteins. This conclusion has been confirmed and extended with the picryl and chlorbenzoyl chloride systems. 1. It is shown that to induce a state of contact sensitivity, the minimal necessary amounts of hapten are of the same order of magnitude, whether this hapten is conjugated with protein or the free reactive chemical itself. From this, it is evident that contamination of conjugates with small amounts of unreacted sensitizer plays no part in the induction of contact reactivity by the conjugate. With the dinitrophenyl system, no contact sensitivity could be induced by the conjugates used; possible reasons for this discrepancy are discussed. 2. Animals sensitized to contact by homologous conjugate can be completely desensitized by injections of such a conjugate in large amount; a similar injection schedule has no effect on the contact sensitivity of animals sensitized with the free reactive sensitizer. 3. The capacity of heterologous (ovalbumin) conjugates to evoke anti-hapten antibodies is shown to be greater than that of homologous (guinea pig seralbumin) conjugates: the reverse is true of their capacity to induce delayed reactivity. 4. Evidence is brought forward to suggest that in animals sensitized with homologous albumin conjugates, the specificity of the delayed reaction involves more than the hapten alone, even though the carrier protein is non-antigenic on its own. The contrast with the apparent lesser specificity of the antibodies later produced is discussed.

Topics: Animals; Antibodies; Antiemetics; Antigens; Carrier Proteins; Dermatitis, Contact; Guinea Pigs; Haptens; Hypersensitivity; Hypersensitivity, Delayed; Ovalbumin; Proteins