ovalbumin and Drug-Hypersensitivity

The Drug Allergy Committee of the Spanish Society of Allergology and Clinical Immunology reviewed the allergenic potential of several substances of food origin that are found in the composition of some drugs. Despite recent legislation on labeling, many labels do not clearly state whether the drug contains raw material (active ingredients, excipient, or other manufacturing intermediate) with an origin in any of the substances in the list of the 14 groups of food allergens that are subject to mandatory declaration. The objective of legislation is that the drug package, the Summary of Product Characteristics, and the patient information leaflet clearly state the food content in order to improve the safety of allergic patients. Therefore, any food or allergen derivative that must be declared should be clearly stated on the drug label. Of all the evaluated products, egg and milk derivatives are the most frequently discussed in literature reviews. The natural or synthetic origin of potentially allergenic substances such as lysozyme, casein, lactose, albumin, phosphatide, and aromatic essences should be clearly stated. Providing this information has 2 clear advantages. First, allergic reactions to drugs in patients with food allergy could be avoided (if the substances have a natural origin). Second, prescription would improve by not restricting drugs containing synthetic substances (which do not usually induce allergic reactions).

Topics: Drug Hypersensitivity; Food Additives; Food Hypersensitivity; Glucosamine; Humans; Lactose; Muramidase; Ovalbumin; Propofol; Spain

The proinflammatory cytokine interleukin-17A (IL-17A) is known to mediate antimicrobial activity, but its role during rhinovirus (RV) infections and in asthma needs further investigation. Therefore, we addressed the role of IL-17A during allergic asthma and antiviral immune response in human and murine immunocompetent cells. In this study we found that asthmatic children with a RV infection in their upper airways have upregulated mRNA levels of the antiviral cytokine interferon type I (IFN)-β and the transcription factor T-box 21 (TBX21) and reduced levels of IL-17A protein in their peripheral blood mononuclear cells (PBMCs). We also found that IL-17A inhibited RV1b replication in infected human lung epithelial cells A549. Furthermore, by using gene array analysis we discovered that targeted deletion of Il17a in murine lung CD4(+) T cells impaired Oas1g mRNA downstream of Ifnβ, independently from RV infection. Additionally, in PBMCs of children with a RV infection in their nasalpharyngeal fluid OAS1 gene expression was found downregulated. Finally RV1b inhibited IL-17A production in lung CD4(+) T cells in a setting of experimental asthma. These results indicate that the RV1b inhibits IL-17A in T helper type 17 cells and IL-17A clears RV1b infection in epithelial cells. In both cases IL-17A contributes to fend off RV1b infection by inducing genes downstream of interferon type I pathway.

Topics: 2',5'-Oligoadenylate Synthetase; A549 Cells; Animals; Asthma; CD4-Positive T-Lymphocytes; Child; Child, Preschool; Drug Hypersensitivity; Female; Gene Expression Regulation; Humans; Interferon-beta; Interleukin-17; Lung; Male; Mice; Mice, Knockout; Ovalbumin; Picornaviridae Infections; Primary Cell Culture; Rhinovirus; RNA, Messenger; Signal Transduction; T-Box Domain Proteins

Allergic airway diseases are accompanied by increased permeability and an inflammatory state of epithelial barriers, which are thought to be susceptible to allergen sensitization. Although exogenous drivers (proteases, allergens) of epithelial barrier disruption and sensitization are well studied, endogenous contributors (diet, xenobiotics, hormones, and metabolism) to allergic sensitization are much less understood. Xenoestrogens are synthetic or natural chemical compounds that have the ability to mimic estrogen and are ubiquitous in the food and water supply of developed countries. By interfering with the estrogen produced by the endocrine system, these compounds have the systemic potential to disrupt the homeostasis of multiple tissues. Our study examined the potential of prototypical xenoestrogen bisphenol A (BPA) to disrupt epithelial homeostasis

Topics: Administration, Inhalation; Animals; Asthma; Benzhydryl Compounds; Cell Line; Cell Proliferation; Cytokines; Drug Hypersensitivity; Endocrine Disruptors; Epithelial Cells; Female; Gene Expression Regulation; Humans; Inflammation; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Phenols; Respiratory Mucosa

Adverse reactions of injectable drugs usually occur at first administration and are closely associated with the dosage and speed of injection. This phenomenon is correlated with the anaphylactoid reaction. However, up to now, study methods based on antigen detection have still not gained wide acceptance and single physiological indicators cannot be utilized to differentiate anaphylactoid reactions from allergic reactions and inflammatory reactions. In this study, a reliable method for the evaluation of anaphylactoid reactions caused by injectable drugs was established by using multiple physiological indicators. We used compound 48/80, ovalbumin and endotoxin as the sensitization agents to induce anaphylactoid, allergic and inflammatory reactions. Different experimental animals (guinea pig and nude rat) and different modes of administration (intramuscular, intravenous and intraperitoneal injection) and different times (15 min, 30 min and 60 min) were evaluated to optimize the study protocol. The results showed that the optimal way to achieve sensitization involved treating guinea pigs with the different agents by intravenous injection for 30 min. Further, seven related humoral factors including 5-HT, SC5b-9, Bb, C4d, IL-6, C3a and histamine were detected by HPLC analysis and ELISA assay to determine their expression level. The results showed that five of them, including 5-HT, SC5b-9, Bb, C4d and IL-6, displayed significant differences between anaphylactoid, allergic and inflammatory reactions, which indicated that their combination could be used to distinguish these three reactions. Then different injectable drugs were used to verify this method and the results showed that the chosen indicators exhibited good correlation with the anaphylactoid reaction which indicated that the established method was both practical and reliable. Our research provides a feasible method for the diagnosis of the serious adverse reactions caused by injectable drugs which could be used in the clinical practice.

Topics: Anaphylaxis; Animals; Biomarkers; Chromatography, High Pressure Liquid; Complement Membrane Attack Complex; Drug Hypersensitivity; Endotoxins; Guinea Pigs; Histamine; Injections, Intravenous; Interleukin-6; Ovalbumin; Rats; Serotonin; Time Factors

The inhalation of manufactured metal oxide nanoparticles may lead to pulmonary toxicity. For instance, ZnO nanoparticles are known to induce pulmonary oxidative stress and inflammation. On the other hand, the pulmonary toxicity of TiO2 nanoparticles is less than that of ZnO nanoparticles. Although, there have been some investigations concerning the induction of pulmonary oxidative stress and inflammation caused by manufactured metal oxide nanoparticles. And, although, it has reported that some nanoparticles cause aggravation of allergic reactions, there have so far been no reports regarding allergy aggravation effects of manufactured metal oxide nanoparticles. In this study, three types of nanoparticles, TiO2, ZnO and SiO2, were administered to mouse lungs by pharyngeal aspiration. Subsequently, the mice inhaled ovalbumin (OVA) a total of eight times over 3 weeks. After inhalation of OVA, the concentrations of total IgE, OVA-specific IgE and OVA-specific IgG1 in serum increased in the mice treated with ZnO. TiO2 and SiO2 nanoparticles did not affect the OVA-specific IgE and IgG1 levels. These results suggest that ZnO nanoparticles have the potential to aggravate allergic reactions. The results also suggest that Zn(2+) release from ZnO nanoparticles is involved in the aggravation potential of allergies. However, pharyngeal aspiration of ZnCl2 solution was not able to aggravate allergic reactions. Continuous Zn(2+) release from ZnO nanoparticles to the lung is necessary for the aggravation of allergic reactions.

Topics: Allergens; Animals; Drug Hypersensitivity; Enzyme-Linked Immunosorbent Assay; Female; Immune Tolerance; Immunoglobulin E; Immunoglobulin G; Inhalation Exposure; Metal Nanoparticles; Mice; Mice, Inbred C57BL; Ovalbumin; Zinc Oxide

This study was designed to investigate the modulatory effects of submicron and nanosized iron oxide (Fe(2)O(3)) particles on the ovalbumin (OVA)-induced immune Th2 response in BALB/c mice. Particles were intratracheally administered four times to mice before and during the OVA sensitization period. For each particle type, three different doses, namely 4×100, 4×250 or 4×500 μg/mouse, were used and for each dose, four groups of mice, i.e. group saline solution (1), OVA (2), particles (3), and OVA plus particles (4), were constituted. Mice exposed to OVA alone exhibited an allergic Th2-dominated response with a consistent increase in inflammatory scores, eosinophil numbers, specific IgE levels and IL-4 production. When the mice were exposed to OVA and to high and intermediate doses of iron oxide submicron- or nanoparticles, the OVA-induced allergic response was significantly inhibited, as evidenced by the decrease in eosinophil cell influx and specific IgE levels. However, the low dose (4×100 μg) of submicron particles had no significant effect on the OVA allergic response while the same dose of nanoparticles had an adjuvant effect on the Th2 response to OVA. In conclusion, these data demonstrate that the pulmonary immune response to OVA is a sensitive target for intratracheally instilled particles. Depending on the particle dose and size, the allergic response was suppressed or enhanced.

Topics: Adaptive Immunity; Animals; Bronchoalveolar Lavage Fluid; Cytokines; Drug Hypersensitivity; Female; Ferric Compounds; Gene Expression Regulation; Immunoglobulin E; Lung; Lung Diseases; Metal Nanoparticles; Mice; Mice, Inbred BALB C; Ovalbumin

An association has been observed between indoor mold contamination and lung allergy and asthma. This relationship is not fully understood. 1→3-β-Glucan is the major cell wall component of fungi and a good marker of fungi exposure. The objective was to evaluate the adjuvant effect of zymosan, a crude yeast cell wall preparation of 1→3-β-glucan, during ovalbumin (OVA) sensitization in an allergy model. BALB/c mice were sensitized by pharyngeal aspiration with saline, 50 μg of OVA, or OVA with 1, 10, 50, or 75 μg of zymosan on days 0, 7, and 14. One week after sensitization, each sensitized animal group was challenged with an aspiration dose of 50 μg of OVA once a week for 2 weeks. At 1 day after the last aspiration, bronchoalveolar lavage fluid and blood was collected, and markers of lung allergy and inflammation were assessed. An adjuvant effect of zymosan on OVA allergy during sensitization was observed as indicated by significant elevations in lung eosinophils, serum OVA-specific IgE, and lung IL-5 in the groups sensitized with zymosan and OVA. Pulmonary treatment with zymosan also amplified lung inflammation. Elevations were observed in lung neutrophils, TNF-α, and parameters of lung injury in the groups primed with both zymosan and OVA. In nearly all parameters, a non-linear dose-response relationship was observed in the groups primed with OVA and zymosan. The optimum adjuvant dose of zymosan was 10 μg. This study demonstrated an adjuvant effect of zymosan when exposures occurred during the sensitization phase in an OVA-induced allergy model in BALB/c mice.

Topics: Adjuvants, Immunologic; Administration, Inhalation; Animals; Asthma; Bronchoalveolar Lavage Fluid; Disease Models, Animal; Dose-Response Relationship, Immunologic; Drug Hypersensitivity; Drug Therapy, Combination; Eosinophils; Immunoglobulin E; Interleukin-5; Male; Mice; Mice, Inbred BALB C; Ovalbumin; Specific Pathogen-Free Organisms; Zymosan

Immune-mediated drug hypersensitivity is a particularly concerning health-safety issue among clinicians given its unpredictability and potentially life-threatening effects, especially with exposure to intravenous drugs. Therefore, the development of intravenous drug-exposure models for drug-hazard assessments has garnered increasing interest in recent years. In this study, we used reporter antigens popliteal lymph node assay to investigate the potential value of intravenous exposure to a selected variety of allergenic compounds, including ovalbumin (OVA), concanavalin A (ConA) and diclofenac. The trinitrophenyl (TNP)-specific antibody-forming cells were used to assess the systemic immune responses to a bystander antigen. Mice were subsequently sensitized by TNP-OVA, and then intravenous exposure to one of the selective compounds. As expected, all positive compounds induced significant popliteal lymph node (PLN) proliferation compared with the control. OVA significantly increased Cluster of Differentiation 4 receptors (CD⁴)⁺ interleukin-4 (IL-4)⁺ T-helper 2 (Th2) cells and, consequently, increased the ratios of IL-4/interferon-γ (IFN-γ) antibody-forming cells (AFCs) in PLNs, while bringing about a dose-dependent increase in immunoglobulin G1 (IgG1) AFCs; these findings indicate that a Th2 hypersensitivity response was induced. A Th2 response was also observed in diclofenac sodium-treated groups, and for ConA, a more mixed Th1/Th2 immune response appeared to be induced. In addition, there was no marked reaction with the negative compound. Together, it seems likely that the intravenous exposure model may be useful for drug-induced systemic hypersensitivity assessments.

Topics: Adjuvants, Immunologic; Allergens; Animals; Antigen-Antibody Reactions; Antigens; Cell Proliferation; Concanavalin A; Diclofenac; Drug Hypersensitivity; Female; Injections, Intravenous; Local Lymph Node Assay; Lymph Nodes; Mice; Mice, Inbred BALB C; Ovalbumin; Risk Assessment; Trinitrobenzenes

Asthma is an allergic lung disease can be modulated by drugs that modify the activity of central nervous system (CNS) such as amphetamine (AMPH). AMPH is a highly abused drug that exerts potent effects on behavior and immunity. In this study we investigated the mechanism involved in the effects of long-term AMPH treatment on the increased magnitude of allergic lung response. We evaluated mast cells degranulation, cytokines release, airways responsiveness and, expression of adhesion molecules. Male Wistar rats were treated with AMPH or vehicle (PBS) for 21 days and sensitized with ovalbumin (OVA) one week after the first injection of vehicle or AMPH. Fourteen days after the sensitization, the rats were challenged with an OVA aerosol, and 24h later their parameters were analyzed. In allergic rats, the treatment with AMPH exacerbated the lung cell recruitment due increased expression of ICAM-1, PECAM-1 and Mac-1 in granulocytes and macrophages recovered from bronchoalveolar lavage. Elevated levels of IL-4, but decreased levels of IL-10 were also found in samples of lung explants after AMPH treatment. Conversely, the ex-vivo tracheal hyper-responsiveness to methacholine (MCh) was reduced by AMPH treatment, whereas the force contraction of tracheal segments due to in vitro antigen challenge remained unaltered. Our findings suggest that lung inflammation and airway hyper-responsiveness due to OVA challenge are under the distinct control of AMPH during long-term treatment. Our data strongly indicate that AMPH positively modulates allergic lung inflammation via the increase of ICAM-1, PECAM-1, Mac-1 and IL-4. AMPH also abrogates the release of the anti-inflammatory cytokine IL-10.

Topics: Amphetamine; Animals; Bronchial Spasm; Bronchoalveolar Lavage Fluid; Cytokines; Drug Administration Schedule; Drug Hypersensitivity; Gene Expression Regulation; Inflammation; Male; Ovalbumin; Rats; Rats, Wistar; Sympathomimetics

Immune-mediated drug hypersensitivity reactions are important causes of black box warnings and drug withdrawals. Despite the high demand for preclinical screening tools, no validated in vitro or in vivo models are available. In the current study, we used a previously described oral administration model using trinitrophenyl-ovalbumin (TNP-OVA) as an antigen to report immuno-adjuvating effects of the analgesic drug acetaminophen (APAP) and its nonhepatotoxic regioisomer 3'-hydroxyacetanilide (AMAP), the antibiotic ofloxacin (OFLX), the antiepileptic drug carbamazepine (CMZ), and the antidiabetic drug metformin (MET). Furthermore, APAP and AMAP were tested in a popliteal lymph node assay (PLNA) combined with TNP-OVA as reporter antigen (RA). C3H/HeOuJ mice were dosed by oral gavage with diclofenac (DF), APAP, AMAP, OFLX, MET, or CMZ. On the first exposure day, the mice received an ip injection with TNP-OVA. Fifteen days later, they were ear challenged with TNP-OVA and delayed-type hypersensitivity (DTH) responses were assessed 24 h later. One week after challenge, the ear-draining lymph node was removed and TNP-specific antibody-secreting cells were determined. DF, APAP, CMZ, and OFLX showed a significant increase in DTH responses to ear injection with TNP-OVA, whereas AMAP and MET did not. C57BL/6 mice were slightly less responsive to APAP and DF after oral gavage, and importantly both AMAP and APAP were negative in the RA-PLNA. The present work shows that the oral exposure model using RA and the RA-PLNA may serve to screen the immune-adjuvant potential of new chemical entities during preclinical drug development.

Topics: Acetaminophen; Acetanilides; Adjuvants, Immunologic; Administration, Oral; Animals; Antibody Formation; Carbamazepine; Drug Evaluation, Preclinical; Drug Hypersensitivity; Female; Injections, Epidural; Local Lymph Node Assay; Metformin; Mice; Mice, Inbred C3H; Mice, Inbred C57BL; Ofloxacin; Ovalbumin

Topics: Drug Hypersensitivity; Enzyme-Linked Immunosorbent Assay; Humans; Influenza A Virus, H1N1 Subtype; Influenza Vaccines; Influenza, Human; Lot Quality Assurance Sampling; Ovalbumin; Seasons; Vaccination

Asthma is a chronic lung disease characterized by allergen-induced airway inflammation and orchestrated by Th2 cells. Interleukin-12, a Th1-promoting cytokine, is capable of inhibit the Th2-driven allergen-induced airway changes and therefore considered as an attractive molecule to treat asthma. Recent epidemiological and clinical studies suggest a possible role of Lactococcus lactis in the prevention of allergic diseases. In this study, we evaluated the immunomodulatory effects of live L. lactis secreting a biologically active form of IL-12 (LL-IL12) in a mouse model of ovalbumin (OVA)-induced asthma. Intranasal mice administration with LL-IL12 resulted in a shift Th2 to Th1 with elevated IFN-gamma and decreased IL-4 levels. In addition, a profound decrease in airway hyper-responsiveness and pulmonary inflammation was also observed in mice administered with LL-IL12. These promising preclinical results suggest the feasibility of this approach to be used in the treatment of asthma.

Topics: Adjuvants, Immunologic; Administration, Intranasal; Animals; Anti-Asthmatic Agents; Asthma; Cytokines; Drug Hypersensitivity; Eosinophils; Interleukin-12; Lactococcus lactis; Lung; Mice; Mice, Inbred BALB C; Ovalbumin; Plasmids; Th1 Cells; Th2 Cells

A nasal formulation of mometasone furoate (MF) is advantageous in avoiding systemic activity characteristic of glucocorticoids when it is applied topically. To confirm antiallergic effects of this glucocorticoid formulation elaborately, we investigated whether the drug can suppress the production of IgE antibodies and related cytokines. It we showed that IgE production induced in mice immunized via intranasal route was significantly reduced when the mice were administered MF intranasally. Further, MF was effective in inhibiting production of type-2 helper T cell cytokines in vivo and in vitro. These results provide a immunopharmacological basis for clinical efficacy of this drug.

Topics: Administration, Intranasal; Animals; Anti-Allergic Agents; Antibody Formation; Cell Line; Drug Hypersensitivity; Immunization; Immunoglobulin E; Immunoglobulin G; Injections, Subcutaneous; Interleukin-4; Male; Mice; Mice, Inbred BALB C; Mometasone Furoate; Nitro Compounds; Ovalbumin; Pregnadienediols; Th2 Cells

The capability of certain drugs to cause immune-mediated drug hypersensitivity reactions in susceptible individuals has initiated a search for pre-clinical screening tools to identify immunosensitizing drugs. Since most drugs are taken orally, hazard assessment of their immunosensitizing potential should include oral exposure models. In this study, the predictive value of the reporter antigen (RA) approach was investigated in combination with oral or intraperitoneal (ip) exposure to a selection of allergenic drugs, i.e., D-penicillamine (D-Pen), Diclofenac (DF), or Nevirapine (Nevi). The RA trinitrophenyl-Ovalbumin (TNP-OVA) was used to assess the capacity of the drugs to stimulate systemic immune responses to a bystander antigen, whereas the RA TNP-Ficoll was used to indicate whether the drugs were able to induce specific anamnestic T-cell responses. TNP-OVA was injected (ip) in C3H/HeOuJ mice that were subsequently exposed (orally or ip) to one of the drugs via different exposure protocols. All three model drugs used resulted in delayed type hypersensitivity reactions to TNP-OVA after ip and oral exposure. In addition, TNP-specific serum antibody levels were increased after ip exposure to Nevi, and after both oral and ip exposure to D-Pen and DF. These data indicate that the present drugs are able to stimulate immune responses to bystander antigens. Responses to TNP-Ficoll were measured in the popliteal lymph node of BALB/c mice three weeks after they received a single oral dose of D-Pen or DF. Results of this approach show that orally pre-treated mice responded with enhanced responses (TNP-specific IgG1 and IFN-gamma production) to sub-optimal doses of D-Pen or DF in a drug-specific manner. Data with TNP-Ficoll indicate that these drugs stimulate systemic formation of specific T cells. Together, the RA-approach allows assessment of systemic sensitization upon oral and/or ip exposure to the selected drugs. To further evaluate the utility of these models, more drugs, including non-allergenic drugs and those that require metabolic conversion to become allergenic need to be studied in the present models.

Topics: Administration, Oral; Allergens; Animals; Antigen-Antibody Reactions; Antigens; Disease Models, Animal; Drug Hypersensitivity; Female; Ficoll; Immunologic Memory; Injections, Intraperitoneal; Lymph Nodes; Mice; Mice, Inbred BALB C; Mice, Inbred C3H; Ovalbumin; Predictive Value of Tests

The use of mAbs to abrogate costimulatory interactions has attracted much attention with regard to prevention and modulation of adverse (auto)immune-like reactions. However, the role of costimulatory molecules and possible therapeutic use of Ab-treatment in drug-induced immunostimulation is poorly elucidated. In the present studies, we show that CD28/CTLA-4-CD80/CD86 costimulatory interactions differently regulate drug-induced type 1 and type 2 responses to an identical bystander Ag, TNP-OVA, in BALB/c mice using the reporter Ag popliteal lymph node assay. The antirheumatic drug D-Penicillamine, which may induce lupus-like side-effects, stimulated type 2 responses against TNP-OVA, characterized by the production of IL-4 and TNP-specific IgG1 and IgE. These responses were abrogated in CD80/CD86-deficient mice and in wild-type mice that were treated with anti-CD80 and anti-CD86, or CTLA-4-Ig. Anti-CTLA-4 intensively enhanced the D-Penicillamine-induced effects. In contrast, the type 1 response (IFN-gamma, TNF-alpha, IgG2a) to TNP-OVA induced by the diabetogen streptozotocin still developed in the absence of CD80/CD86 costimulatory signaling. In addition, it was demonstrated that coadministration of anti-CD80 and anti-CD86 mAbs slightly enhanced streptozotocin-induced type 1 responses, whereas the CTLA-4-Ig fusion protein completely abrogated this response. In conclusion, different drugs may stimulate distinct types of immune responses against an identical bystander Ag, which are completely dependent on (type 2) or independent of (type 1) the CD28/CTLA-4-CD80/CD86 pathway. Importantly, the effects of treatment with anti-CD80/CD86 mAbs and CTLA-4-Ig may be considerably different in responses induced by distinct drugs.

Topics: Animals; Antibodies; Antigens, CD; Antigens, Differentiation; B7-1 Antigen; Bystander Effect; CD28 Antigens; CTLA-4 Antigen; Drug Hypersensitivity; Female; Immunity; Mice; Mice, Inbred BALB C; Mice, Knockout; Ovalbumin; Penicillamine; Signal Transduction; Streptozocin; Th1 Cells; Th2 Cells

We detected a human humoral peptide that deglycosylates mouse antibody IgE, and found that this peptide had the amino acid sequence ADSGEGDFLAEGGGV. The peptide synthesized according to the sequence also deglycosylated the antibody IgE. The deglycosylated IgE did not induce mouse systemic anaphylaxis. Human fibrinopeptide A has the amino acid sequence indicated above, and a polypeptide extracted from human urine showed an antiallergic effect on humans and mice, which strongly suggests that fibrinopeptide A mediates allergic reaction via antibody IgE-deglycosylation, and is excreted as a polypeptide in urine.

Topics: Acute-Phase Reaction; Adult; Amino Acid Sequence; Animals; Drug Hypersensitivity; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Female; Fibrinopeptide A; Glycosylation; Glycosyltransferases; Humans; Hypersensitivity; Immune Sera; Immunoglobulin E; Injections, Intraperitoneal; Male; Mice; Mice, Inbred Strains; Molecular Sequence Data; Ovalbumin; Peptide Fragments; Structure-Activity Relationship

The effect of lipids administration by gavage (0.4% body weight) given daily during four weeks on the hypersensitivity reaction in trachea, upper and lower bronchi, liver, kidney, mesentery, and pancreas was investigated in male rats. The plasma exudation was assessed by using Evans blue (EB) dye extravasation method. There was a significant difference in the permeability of the organs in nonimmunized rats. The immunization increased the vascular permeability and the response with the organs varied greatly. The effect of lipids on anaphylactic reaction was compared to those of untreated rats (control group). The EB extravasation was significantly increased in the trachea obtained from rats treated with cocoa butter and soybean oil. In the upper bronchi of rats treated with soybean oil, the EB extravasation was increased. However, in the lower bronchi, none of the treatments with lipids changed the extravasation of EB. The same was observed in the liver and kidney. The animals treated with lipids by gavage did not present differences in EB extravasation in the mesentery. However, in the pancreas and duodenum, the treatment with fish and soybean oils and cocoa butter markedly lowered EB extravasation.

Topics: Administration, Oral; Anaphylaxis; Animals; Capillary Permeability; Dietary Fats; Drug Hypersensitivity; Evans Blue; Immunization; Male; Ovalbumin; Rats

No adequate enteral sensitization models are available to study food allergy and allergenicity of food proteins. Using a previously described oral sensitization protocol to sensitize Brown Norway rats (BN) to food proteins, the influence of genetically-based strain-specific characteristics of the immune system on the outcome of oral sensitization studies was investigated. BN, Hooded Lister (HL), Piebald Virol Glaxo (PVG) and Wistar rats were daily administered 1 mg of ovalbumin (OVA) by gavage dosing for 42 days without the use of an adjuvants. The highest OVA-specific IgG responses were detected in the BN rats followed by Wistar, HL and PVG rats. OVA-specific IgE responses were only detectable in the BN rats. The cellular immune response was examined by determination of delayed-type hypersensitivity (DTH) reactions in the animals. The response was most pronounced in the HL and Wistar rats. PVG and BN rats showed comparable DTH responses but the responses were significantly weaker than those observed in HL and Wistar rats. It was concluded that the genetic make-up of different rat strains influences the outcome of oral sensitization studies. In addition, using the described oral sensitization protocol, the BN rat seems to be the most suitable strain for inducing oral sensitization.

Topics: Administration, Oral; Animals; Antibody Formation; Drug Hypersensitivity; Hypersensitivity, Delayed; Immunity, Cellular; Immunoglobulin E; Immunoglobulin G; Male; Ovalbumin; Rats; Rats, Inbred BN; Rats, Wistar; Species Specificity

The cytokines IL-4, IL-5, and IL-13, secreted by Th2 cells, have distinct functions in the pathogenesis of asthma. We have previously shown that the transcription factor GATA-3 is expressed in Th2 but not Th1 cells. However, it was unclear whether GATA-3 controls the expression of all Th2 cytokines. Expression of a dominant-negative mutant of GATA-3 in mice in a T cell-specific fashion led to a reduction in the levels of all the Th2 cytokines IL-4, IL-5, and IL-13. Airway eosinophilia, mucus production, and IgE synthesis, all key features of asthma, were severely attenuated in the transgenic mice. Thus, targeting GATA-3 activity alone is sufficient to blunt Th2 responses in vivo, thereby establishing GATA-3 as a potential therapeutic target in the treatment of asthma and allergic diseases.

Topics: Aerosols; Amino Acid Substitution; Animals; Asthma; Bronchoalveolar Lavage Fluid; DNA-Binding Proteins; Drug Hypersensitivity; Eosinophilia; GATA3 Transcription Factor; Gene Expression Regulation; Genes, Dominant; Immunization; Immunoglobulin E; Inflammation; Interleukin-13; Interleukin-4; Interleukin-5; Lung; Mice; Mice, Inbred C57BL; Mice, Transgenic; Mucus; Mutagenesis, Site-Directed; Ovalbumin; Th2 Cells; Trans-Activators

In an attempt to study the pathogenesis of mucosal hypersensitivity in allergic rhinitis, we investigated the suppressive effects of cyclosporin A (CyA) and glucocorticosteroids on ovalbumin (OA)-induced hypersensitivity to topical histamine challenge.. Actively sensitized Dunkin-Hartley guinea pigs.. OA and alum were applied to guinea pigs intraperitoneally 3 times at two-week intervals. After general sensitization, OA inhalation was performed every day for 6 days as topical sensitization. Before inhalation, treatment with CyA (50 mg/kg, p.o.), glucocorticosteroids (beclomethasone propionate (1.0 mg/kg, i.p.), fluticasone propionate (FP, 0.5 mg/kg, i.p.)) or vehicle were performed, and the sensitivity to histamine was measured before and after the inhalation. Moreover, in actively (general and topical) sensitized guinea pigs, FP (0.5 mg/kg, i.p.) was applied every day for 5 days and histamine sensitivity was evaluated before and after the application.. We found that histamine sensitivity was significantly increased by nasal antigen challenge in this guinea pig model, and that the occurrence of histamine hypersensitivity was inhibited by the pretreatment with CyA and glucocorticosteroids. Although multiple administration of FP gradually reduced the histamine hypersensitivity according to the period of administration, it did not significantly alter the histamine hypersensitivity after the occurrence of hypersensitivity.. It is concluded that CyA and glucocorticosteroids suppress antigen-induced histamine hypersensitivity in a guinea pig model of allergic rhinitis.

Topics: Administration, Inhalation; Androstadienes; Animals; Anti-Allergic Agents; Anti-Inflammatory Agents; Beclomethasone; Cyclosporine; Disease Models, Animal; Drug Hypersensitivity; Fluticasone; Glucocorticoids; Guinea Pigs; Histamine; Immunosuppressive Agents; Injections, Intraperitoneal; Male; Ovalbumin; Rhinitis, Allergic, Perennial

Eotaxin administration intraperitoneally, but not into dorsal air-pouches, of ovalbumin-sensitized mice exhibiting blood eosinophilia induced a threefold increase in eosinophil (E phi s) infiltration. Transfer of 1 x 10(6) mixed peritoneal cavity cells (PCC), containing 3.5 to 4.5 x 10(4) mast cells (MC), from donor mice to air-pouches of sensitized (but not unsensitized) recipient mice, established an E phi infiltration to eotaxin (vehicle, 0.86 +/- 0.27 x 10(6); eotaxin, 1.63 +/- 0.16 x 10(6) E phi s/air-pouch). Neutrophil numbers were also increased. When MC-depleted (-93%) PCC were injected into air-pouches of recipient animals, E phi infiltration was not supported (-52%). Injection of macrophage-depleted (-99%) PCC into air-pouches elicited a full E phi response to eotaxin but not neutrophil infiltration (-81%). Systemic dexamethasone treatment of recipient mice reduced E phi accumulation; treatment of donor mice only reduced neutrophil accumulation. Our study points to a crucial role for MC in E phi recruitment by eotaxin.

Topics: Animals; Anti-Inflammatory Agents; Cell Line; Cell Movement; Chemokine CCL11; Chemokines, CC; Chemotactic Factors, Eosinophil; Cytokines; Dexamethasone; Drug Hypersensitivity; Eosinophilia; Eosinophils; Female; Leukocyte Count; Macrophages; Mast Cells; Mice; Mice, Inbred BALB C; Neutrophils; Ovalbumin; Peritoneal Cavity; Recombinant Proteins; Skin

The purpose of this study was to determine if the sustained release of ampicillin from a biodegradable drug-delivery system (microencapsulated ampicillin anhydrate (MEAA)) will increase or decrease the intensity of a hypersensitivity reaction compared with that observed with free drug. Ovalbumin, which is known to elicit a marked hypersensitivity reaction in guinea pigs, and microencapsulated ovalbumin (MOVA) were tested in parallel with ampicillin and MEAA. Guinea pigs were sensitized biweekly by subcutaneous and intramuscular injections of ampicillin, MEAA, ovalbumin, MOVA or placebo microspheres (test articles), each mixed with Freund's adjuvant, and challenged 2 weeks later, intradermally, with the free compounds. In a separate set of experiments, guinea pigs were sensitized by implantation of the same agents in the caudal thigh of anaesthetized animals. Skin allergic reactions were tested at 1 and 3 weeks following local implantation of the test articles. Sera of sensitized guinea pigs were tested for specific IgG antibodies by enzyme-linked immunosorbent assay, and skin samples from the site of the inflammatory reaction were fixed, stained and evaluated histologically. Guinea pigs sensitized systemically with MEAA or MOVA showed smaller, but not statistically different skin allergic response than animals given corresponding free compounds. However, guinea pigs sensitized by local implantation of MEAA showed a significantly lower inflammatory response (P < 0.0001) than those given an equivalent dose of the free drug. Guinea pigs sensitized with placebo microspheres showed a low inflammatory skin reaction which was similar to those sensitized with all doses of MEAA. There was no significant difference in specific IgG antibody response in the sera of guinea pigs sensitized locally with either free or microencapsulated ampicillin or ovalbumin. Histology of skin revealed a milder inflammatory reaction with MEAA or MOVA than with ampicillin or ovalbumin, respectively. We conclude that the encapsulated ampicillin or ovalbumin and subsequent release of each agent will elicit a reduced hypersensitivity reaction in guinea pigs than will the free agent.

Topics: Ampicillin; Animals; Antibiotic Prophylaxis; Antibodies; Antibody Specificity; Capsules; Drug Hypersensitivity; Drug Implants; Female; Guinea Pigs; Inflammation; Male; Ovalbumin; Penicillins

To study the role of tachykinins in allergic responses in the airways of guinea pigs sensitized to ovalbumin (OVA), we examined the bronchial contractile response to allergen in the presence or absence of the tachykinin antagonist FK224 in vitro. Because neutral endopeptidase (NEP) effectively cleaves tachykinins, we incubated bronchial tissues with the NEP inhibitor phosphoramidon (10(-5) M) to maintain the activity of endogenously released tachykinins. Then we added 10(-5)% (10 microns/ml) OVA in the presence or absence of FK224 (10(-5) M). FK224 significantly inhibited OVA-induced contraction plateaued and began to relax, we added 10(-5) M phosphoramidon. In the tissue without FK224, phosphoramidon blocked the relaxation and enhanced the contraction. In contrast, in the tissues treated with FK224, phosphoramidon did not enhance the OVA-induced contraction. The enhancement of the contraction induced by phosphoramidon was not inhibited by the sodium channel blocker tetrodotoxin. These results suggest that (1) allergic response causes release of tachykinin-like substances to induce bronchial contraction in part, (2) these responses are blocked by tachykinin antagonist FK224 and (3) nerve conduction is not necessary for the release of tachykinin-like substances induced by allergic response in the guinea pig bronchus.

Topics: Allergens; Animals; Bronchoconstriction; Drug Hypersensitivity; Glycopeptides; Guinea Pigs; Male; Metalloendopeptidases; Ovalbumin; Peptides, Cyclic; Protease Inhibitors; Tachykinins; Tetrodotoxin

To determine the role of leukotriene (LT)-degrading enzymes in allergic reactions, we studied the effects of inhibitors of gamma-glutamyl transpeptidase (gamma-GTP) and dipeptidases on increases in pulmonary insufflation pressure (PIP) and vascular permeability induced by ovalbumin (OA) antigen in guinea pigs sensitized to OA antigen in vivo. Vascular permeability was assessed by the amount of extravasated Evans blue dye from the trachea, main bronchi, and segmental bronchi. An intravenous (i.v.) administration of OA antigen (200 micrograms/kg) caused increases in PIP and extravasated Evans blue dye, and OA antigen-induced effects were potentiated by gamma-GTP inhibitor L-serine borate (3 x 10(-5) M/kg, i.v.) (P < 0.05) and an inhibitor of dipeptidases, L-cysteine (3 x 10(-5) M/kg, i.v.) (P < 0.01). OA antigen-induced increases in PIP and Evans blue dye extravasation were in part inhibited by LT-receptor antagonist ONO-1078 (10(-4) M/kg, i.v.). Guinea-pig tracheal tissues contained gamma-GTP and microsomal dipeptidase activities. Histochemical and immunohistochemical studies indicate that gamma-GTP-like activity existed in the epithelium and smooth muscle, and an activity of microsomal dipeptidase was observed in the endothelial cells of microvessels and epithelium. These results suggest that LT-degrading enzymes have an important role in regulating allergic reaction in the airway in vivo.

Topics: Animals; Capillary Permeability; Dipeptidases; Dose-Response Relationship, Drug; Drug Hypersensitivity; Evans Blue; gamma-Glutamyltransferase; Glycosylphosphatidylinositols; GPI-Linked Proteins; Guinea Pigs; Leukotriene C4; Leukotriene D4; Leukotriene E4; Leukotrienes; Lung; Male; Muscle, Smooth; Ovalbumin; Trachea

Asthma is considered to be a chronic inflammatory response of the airways characterized by a leukocyte infiltration into the lungs. Whereas lymphocytes and macrophages are involved in the initiation and propagation of inflammation, both neutrophils and in particular eosinophils are considered to play major effector roles. Therefore, allergic animal models in various species have been established to assess leukocyte infiltration by bronchoalveolar lavage (BAL) of antigen-sensitized and antigen-challenged animals as an inflammatory parameter in asthma pharmacology. Differential leukocyte counts in BAL fluids are routinely assessed by visual microscopic analysis of stained slides after cytocentrifugation. This procedure is very time-consuming, and the underlying standard morphological criteria may vary between different observers. In the present paper, we propose an alternative automatic method for leukocyte differentiation in BAL fluids from ovalbumin-treated guinea pigs and Brown-Norway rats using Cobas Helios 5Diff from Hoffmann-La Roche. BAL samples are directly applied to the analyzer and are automatically mixed with "Eosinofix," which stabilizes leukocyte membranes and specifically stains eosinophils. By a combination of electric (resistance) and optical (light scatter) analysis, the lymphocytes, monocytes/macrophages, neutrophils, and eosinophils are discriminated and the total leukocyte numbers are obtained. For both animal species we found high correlations for all leukocyte populations by comparing the results obtained with Cobas Helios 5Diff and conventional microscopic analysis. The major advantage of the automatic method is the much lower (about one-third) time requirement.

Topics: Animals; Bronchoalveolar Lavage Fluid; Coloring Agents; Drug Hypersensitivity; Guinea Pigs; Leukocyte Count; Male; Ovalbumin; Rats; Serine Proteinase Inhibitors

Formaldehyde (FA), a common indoor air pollutant, has been associated with increased prevalence rates of asthmatic symptoms among exposed individuals in epidemiologic surveys. We studied the influence of FA exposure on inhalative allergic sensitization in the guinea pig. Three groups of guinea pigs (n = 12 each) were exposed to clean air or two different FA concentrations (0.13 and 0.25 ppm) over 5 consecutive days. Exposure was followed by inhalation of 0.5% ovalbumin (OA) as sensitizing allergen. Three weeks later, specific bronchial provocation with OA was performed with body plethysmographic measurement of compressed air (CA). Furthermore, specific anti-OA-IgGl (reaginic) antibodies were determined in serum. In a further six animals, the respiratory tract was examined histologically for signs of inflammation directly after the end of FA or clean air exposure. In the group exposed to 0.25 ppm FA, 10/12 animals were found to be sensitized to OA (positive reaction on specific provocation) vs. 3/12 animals in the control group (P < 0.01). Furthermore, CA measurements of specific bronchial provocation and serum anti-OA-antibodies were significantly higher in the 0.25 ppm FA group than in controls (CA 0.35 vs. 0.09 ml median, P < 0.01; anti-OA-IgGl 13 vs. < 10 EU median, P < 0.05), indicating enhanced sensitization. In the group exposed to 0.13 ppm FA, no significant difference was found compared to the control group. There was no sign of inflammation of the lower airways in FA-exposed guinea pigs other than mucosal edema, which was discovered by morphometry. We conclude that short-term exposure to a low concentration of FA (0.25 ppm) can significantly enhance sensitization to inhaled allergens in the guinea pig.

Topics: Allergens; Animals; Antibodies; Antibody Specificity; Drug Hypersensitivity; Formaldehyde; Guinea Pigs; Immunization; Mucous Membrane; Ovalbumin

The pharmacological mechanisms of allergic cough in the guinea pig were studied. Actively sensitized guinea pigs were exposed to aerosols of antigen to elicit coughing. In separate experiments, naive guinea pigs were exposed to aerosols of capsaicin to elicit coughing. Both allergic and capsaicin-induced cough were inhibited by loratadine (0.3-10 mg kg-1 p.o.) and chlorpheniramine (0.1-3.0 mg kg-1 p.o.). Neither cimetidine (10 mg kg-1 s.c.), nor thioperamide (3-10 mg kg-1 s.c.), inhibited allergic or capsaicin-induced cough. Codeine (3-30 mg kg-1 p.o.), salbutamol (0.003-3.0 mg kg-1 s.c.) and ipratropium (0.03-1.0 mg kg-1 s.c.) inhibited both allergic and capsaicin-induced cough. Hexamethonium (10 and 30 mg kg-1 s.c.) inhibited allergic, but not capsaicin-induced cough. Allergic and capsaicin-induced cough were unaffected by phenidone (5.0 and 10.0 mg kg-1 s.c.). Indomethacin (5.0 and 10.0 mg kg-1 s.c.) had no effect on allergic cough but slightly inhibited capsaicin-induced cough. We conclude that allergic and capsaicin-induced cough are modulated by histamine H1 receptor and cholinergic mechanisms. Histamine H2 or histamine H3 receptor mechanisms, and lipoxygenase and cyclooxygenase products of arachidonic acid metabolism do not influence allergic and capsaicin-induced cough. Ganglionic mechanisms play a minor role in the production of allergic cough and no role in capsaicin-induced cough.

Topics: Administration, Oral; Aerosols; Albuterol; Analysis of Variance; Animals; Antitussive Agents; Capsaicin; Chlorpheniramine; Cimetidine; Codeine; Cough; Drug Hypersensitivity; Guinea Pigs; Hexamethonium; Histamine Antagonists; Indomethacin; Injections, Subcutaneous; Ipratropium; Loratadine; Male; Ovalbumin; Piperidines; Receptors, Histamine H1; Receptors, Histamine H2

The aims of these studies were to examine the effects of FCC5 (2-carboxamidino-1,2,3,4,10,14b-hexahydrodibenzo (c,f) pyrazino (1,2,-a) azepine HCl), an analogue of mianserin, on immediate type hypersensitivity reactions in-vitro. The actions of FCC5 were examined on the Schultz-Dale reaction of guinea-pig ileum and on histamine and leukotriene release from human- and guinea-pig-sensitized lung fragments. FCC5 (applied topically) was assessed for anti-inflammatory activity in-vivo against phorbol-12-myristate-13-acetate (PMA)-induced oedema in the mouse ear. FCC5 (IC50 = 0.17 microM) was a potent inhibitor of the Schultz-Dale reaction in-vitro, as assessed by a concentration-dependent attenuation of egg albumin-induced contractions of sensitized guinea-pig isolated ileum. Using human and guinea-pig isolated sensitized lung fragments, FCC5 (1-100 microM) attenuated antigen-induced release of sulphidopeptidoleukotrienes and histamine. FCC5 (50 micrograms topically) resembled mianserin and indomethacin in attenuating PMA-induced mouse ear inflammation. These properties together with previously published evidence of long lasting antihistamine properties in-vivo, suggest that FCC5 has therapeutic potential as an anti-allergic agent, especially in pathological conditions where an inflammatory component is present.

Topics: Analysis of Variance; Animals; Anti-Inflammatory Agents, Non-Steroidal; Disease Models, Animal; Drug Hypersensitivity; Drug Interactions; Guinea Pigs; Histamine H1 Antagonists; Histamine Release; Humans; Ileum; Injections, Intraperitoneal; Leukotrienes; Lung; Male; Mianserin; Mice; Muscle Contraction; Muscle, Smooth; Ovalbumin; Phthalazines; Serotonin Antagonists; Tetradecanoylphorbol Acetate

In this study, the effect of soluble IL-4 receptors (sIL-4R) on murine allergen-induced IgE and IgG1 production was examined. Lymphocytes from mice sensitized to the allergens ragweed (RW) or ovalbumin (OVA) in vivo were restimulated in vitro with the sensitizing allergen in the presence of either a soluble murine sIL-4R, a dimeric sIL-4R Ig fusion protein (sIL-4R/Fc), or anti-IL-4 antibody in 14-day cultures. Both monomeric and dimeric sIL-4R inhibited polyclonal IgE (approximately 70%) and IgG1 (approximately 35%) production in a dose-dependent fashion, similar to that observed in the presence of the anti-IL-4 antibody. Allergen-specific IgE and IgG1 were inhibited to a greater degree. Addition of sIL-4R was most effective when present in the culture during the first 3 days and added not later than day 6. In kinetic experiments, we distinguished ongoing IgE production from precommitted B cells versus newly induced IgE synthesis and found that newly induced IgE production was the major target of the sIL-4Rs. These data demonstrate the efficacy of sIL-4R in inhibiting the early stages of the IgE B-cell maturation pathway and indicate the potential of sIL-4R for the inhibition of IgE production in vivo.

Topics: Animals; Antibodies; Base Sequence; Cells, Cultured; DNA, Complementary; Drug Hypersensitivity; Female; Immunoglobulin E; Immunoglobulin G; Lymph Nodes; Mice; Molecular Sequence Data; Ovalbumin; Receptors, Interleukin; Receptors, Interleukin-4; Spleen

A murine model for antigen-induced bronchial hyperreactivity (BHR) and airway eosinophilia, two hallmarks of asthma, was developed using ovalbumin-immunized mice, which produce large amounts of IgE (named BP2, "Bons Producteurs 2," for High Line of Selection 2). A single intranasal ovalbumin challenge failed to modify the bronchial responses, despite the intense eosinophil recruitment into the bronchoalveolar lavage fluid and airways. When mice were challenged twice a day for 2 days or once a day for 10 days, BHR in response to i.v. 5-hydroxytryptamine or to inhaled methacholine was induced in BP2 mice but not in BALB/c mice. Histological examination showed that eosinophils reached the respiratory epithelium after multiple ovalbumin challenges in BP2 mice but remained in the bronchial submucosa in BALB/c mice. Total IgE titers in serum were augmented significantly with immunization in both strains, but much more so in BP2 mice. Interleukin 5 (IL-5) titers in serum and bronchoalveolar lavage fluid of BP2 mice were augmented by the antigenic provocation, and a specific anti-IL5 neutralizing antibody suppressed altogether airway eosinophilia and BHR, indicating a participation of IL-5 in its development. Our results indicate that the recruitment of eosinophils to the airways alone does not induce BHR in mice and that the selective effect on BP2 mice is related to their increased IgE titers associated with antigen-driven eosinophil migration to the epithelium, following formation and secretion of IL-5.

Topics: Animals; Antigens; Bronchi; Drug Hypersensitivity; Eosinophils; Epithelium; Immunoglobulin E; Interleukin-5; Male; Methacholine Chloride; Mice; Mice, Inbred BALB C; Mice, Inbred Strains; Mucous Membrane; Ovalbumin; Serotonin

The aim of this study was to investigate whether in vitro exposure to NO2 affects responsiveness in ovalbumin-sensitized guinea-pig bronchi. Twenty-three animals were sensitized by three weekly intraperitoneal injections of 1 mg ovalbumin in saline with Freund's adjuvant; twenty-one control guinea-pigs received the diluent alone. From each animal, the two main bronchi were obtained and cannulated, then exposed in vitro to a constant intraluminal flow of: (i) either air or 2.5 ppm NO2 with four spikes of 10 ppm NO2 for 2 h; (ii) either air or 10 ppm NO2 for 4 h. A bronchial ring obtained from each animal before exposure was kept in aerated Krebs-Henseleit solution. Rings from bronchi exposed to air, NO2, or kept in Krebs solution were studied isometrically. We performed overall and non-adrenergic non-cholinergic voltage-response curves to electrical field stimulation, concentration-response curves to acetylcholine and to neurokinin A, followed by administration of 10 mg/ml ovalbumin. We did not find any significant difference in bronchial smooth muscle responsiveness between nonexposed, air-exposed and NO2-exposed bronchi, as well as between bronchi from control and sensitized animals. We conclude that in vitro exposure to NO2 does not alter bronchial smooth muscle responsiveness to either specific or non-specific stimuli.

Topics: Acetylcholine; Air Pollutants; Animals; Bronchi; Dose-Response Relationship, Drug; Drug Hypersensitivity; Electric Stimulation; Guinea Pigs; Male; Muscle Contraction; Muscle, Smooth; Neurokinin A; Nitrogen Dioxide; Ovalbumin

The ovalbumin (OA)-sensitized Brown Norway rat (BN) demonstrates early-response (ER) and late-response (LR) allergic bronchoconstriction. To determine whether these responses could be replicated in vitro, we studied lung explants from 8-wk-old male BN rats (wt: 239 +/- 28 g), of which 19 were sensitized to OA (test) and 16 served as controls. Two weeks after sensitization, the animals' lungs were removed, filled with a 1% (wt/vol) agarose-containing solution at 37 degrees C, and cooled to 4 degrees C. Transverse slices (0.5 to 1.0 mm thick) were cut and cultured overnight. Airways were visualized with an inverted microscope and baseline images were obtained with a video camera. To study the ER, 40 airways from 15 test rats and 29 airways from 10 control rats were challenged with 2 micrograms OA and imaged each minute for 10 min. To study the LR, 40 airways from 12 test rats and 44 airways from 12 control rats were challenged with 2 micrograms OA and imaged each hour for 8 h. The maximal response (MR) for each airway was defined as the percent of airway closure. The ER and LR were both defined as an MR > or = mean + 2 SD of the controls. An ER occurred in 38 of 40 test and 2 of 29 control airways (mean MR: 42 +/- 24% versus 4 +/- 3%, p < 0.001), and was completely blocked by methysergide pretreatment in 13 airways.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Airway Resistance; Animals; Asthma; Bronchial Provocation Tests; Bronchodilator Agents; Constriction, Pathologic; Disease Models, Animal; Drug Hypersensitivity; Hypersensitivity, Delayed; Hypersensitivity, Immediate; Immunoglobulin E; In Vitro Techniques; Leukotriene D4; Male; Methysergide; Ovalbumin; Premedication; Propionates; Quinolines; Rats; Serotonin; Time Factors

Immunohistochemical study with antisera to chromogranin A and neuron-specific enolase, a general marker for nerves and endocrine cells, was used to quantify changes in bronchial neuroendocrine cells in guinea-pigs sensitized and challenged with ovalbumin. Actively sensitized animals were killed 2, 6, 24, 48, 72, and more than 144 hours after being challenged by an aerosolized solution of ovalbumin. The number of chromogranin A-immunoreactive cells was significantly greater in sensitized but unchallenged animals and in sensitized animals killed 2 and 6 h after challenge when compared to controls; it decreased significantly in animals killed more than 24 h after challenge when compared to sensitized, unchallenged animals. The number of neuron-specific-enolase-immunoreactive cells did not change. We conclude that the peptide content of bronchial neuroendocrine cells increases during sensitization and in the early phase of a hypersensitivity reaction, and that the cells release their granule contents in the late phase of such a reaction. They may therefore play a role in immunoallergic events in the lung.

Topics: Animals; Biomarkers; Bronchi; Chromogranin A; Chromogranins; Cytoplasmic Granules; Drug Hypersensitivity; Guinea Pigs; Immunization; Male; Neurosecretory Systems; Ovalbumin; Phosphopyruvate Hydratase; Respiratory Hypersensitivity

Selective platelet-activating factor (PAF) antagonists and autodesensitization to this lipid were used to investigate the role of PAF in antigen-induced pleurisy in the rat. Pleural inflammation was triggered by the intrathoracic (i.t.) injection of ovalbumin (12 micrograms/cavity) into animals actively sensitized 14 days before. Successive daily i.t. injections of PAF (1 microgram/cavity) led to selective autodesensitization, which was apparent after the third injection and maximal after the fifth. The PAF antagonists BN 52021 and WEB 2086 inhibited the late pleural eosinophil accumulation caused by antigen but, as also noted with WEB 2170, failed to modify the early antigen-induced plasma exudation and leukocyte infiltration. In contrast to the antagonists, desensitization to PAF was clearly effective against these early alterations. To further investigate this discrepancy, the antigenic challenge was performed 24 h after a single prestimulation with PAF, when sensitivity to the lipid was still intact. Under this condition, plasma exudation and cellular influx triggered by the antigen were also abrogated, indicating that this protective effect was accounted for by a mechanism other than refractoriness to PAF. Because 24 h after PAF injection only eosinophil counts remained elevated, an alternative eosinophilotactic substance was used to further study the mechanism of PAF versus antigen-induced pleural inflammation. Prior treatment with the peptide Ala-Gly-Ser-Glu (ECF-A, 20 micrograms/cavity) also inhibited the allergic pleurisy, whereas the noneosinophilotactic substances histamine (200 micrograms/cavity) and serotonin (100 micrograms/cavity) were inactive. Furthermore, drugs that share the ability to impair PAF-induced eosinophilia, including azelastine and cetirizine, prevented the inhibitory effect of PAF on the antigen-induced pleurisy. These findings suggest that PAF may account for the late eosinophilia, but not for the acute phase of the rat allergic pleurisy, which is clearly attenuated by PAF or ECF-A pretreatment.

Topics: Analysis of Variance; Animals; Azepines; Chemotactic Factors, Eosinophil; Diterpenes; Drug Hypersensitivity; Eosinophils; Female; Freund's Adjuvant; Ginkgolides; Inflammation; Lactones; Leukocyte Count; Male; Oligopeptides; Ovalbumin; Platelet Activating Factor; Platelet Membrane Glycoproteins; Pleurisy; Rats; Rats, Wistar; Receptors, Cell Surface; Receptors, G-Protein-Coupled; Triazoles

Sensitization and challenge of guinea-pig airways with aerosols of ovalbumin solution administered through an endotracheal tube leads to a significant increase in kallikrein-like enzyme activity in bronchoalveolar lavage (BAL) fluid. This increase is present up to six hours after challenge and may be related to changes in airways resistance that persist for similar periods.

Topics: Aerosols; Animals; Bronchoalveolar Lavage Fluid; Drug Hypersensitivity; Guinea Pigs; Kallikreins; Ovalbumin; Reference Values; Respiratory System

The effect of NZ-107 (4-bromo-5-(3-ethoxy-4-methoxybenzylamino)-3(2H)-pyridazinone) on late-phase airway responses and airway hyperreactivity was investigated in the guinea pig. Challenge with inhaled ovalbumin in conscious guinea pigs actively sensitized with inhaled ovalbumin caused triphasic bronchial obstruction, which peaked at 5-30 min, 6-8 h and 24 h. In this model, airway hyperreactivity to acetylcholine was observed 48 h after antigen challenge. Orally administered NZ-107, given 2 h before ovalbumin challenge significantly inhibited airway responses at 5-30 min (10 mg/kg), 6-8 h (30 mg/kg), 24 h (10 mg/kg) and airway hyperreactivity (30 mg/kg). When NZ-107 (10 mg/kg) was orally administered to the guinea pigs 3 h after ovalbumin challenge, it also inhibited airway responses at 6-8 h and 24 h and airway hyperreactivity. In anaesthetized guinea pigs, intravenous administration of NZ-107 (0.03-1.0 mg/kg) inhibited platelet-activating factor (PAF)- and propranolol-induced airway hyperreactivity to histamine. These results suggest that NZ-107 may be a useful drug for the treatment of bronchial asthma by reducing late-phase airway responses and airway hyperreactivity.

Topics: Aerosols; Animals; Antigens; Bronchial Hyperreactivity; Bronchoconstriction; Drug Hypersensitivity; Guinea Pigs; Histamine; Male; Ovalbumin; Platelet Activating Factor; Propranolol; Pyridazines; Respiratory Hypersensitivity

Comparative studies on immunogenicity for compounds (TNBS, CET and 4-AAP) and compound-protein conjugates (TNBS-OA, CET-OA and 4-AAP-OA) were undertaken by the assay systems of humoral reaction (HA) and cellular reaction (DTH) in rabbits. The immunogenicity of TNBS-OA was substantially different from either that of CET-OA or 4-AAP-OA in the assay system of DTH. That is, rabbits immunized with TNBS-OA plus FCA displayed positive DTH reactions to the eliciting antigen (TNBS), while such reactions were never recognized in the rabbits immunized with CET-OA plus FCA or 4-AAP-OA plus FCA to the eliciting antigen of CET or 4-AAP, respectively. The above negative reactions observed in both groups, CET-OA and 4-AAP-OA, can be explained by the ordinary interpretation that the specificity of cellular immunoreaction (DTH) is directed toward the binding part of hapten-protein conjugate. By contrast, TNBS, spread on the skin to evoke DTH, may react with epidermal proteins in vivo to form 2,4,6-trinitrophenyl (TNP)-coupled protein. Consequently, it was suggested that this TNP-coupled protein evoked the positive DTH reaction in the TNBS-OA group.

Topics: Ampyrone; Animals; Cephalothin; Drug Hypersensitivity; Hypersensitivity, Delayed; Male; Ovalbumin; Rabbits; Serum Albumin, Bovine; Trinitrobenzenesulfonic Acid

Antigenicity of terazosin was studied in the experimental animals and the following results were obtained. Terazosin was shown to be non-immunogenic in guinea-pigs and mice when immunized alone or with mixture of terazosin and protein as immunogen. Protein-conjugate of terazosin induced responses of hapten specific antibody when guinea pigs and mice were immunized. However, terazosin alone was shown to be not capable of eliciting any allergic responses. In conclusion, terazosin lacks immunogenicity and eliciting antigenicity in these experimental conditions and this suggests that drug allergic response would either not occur or be minimal, if any, when terazosin is administered clinically.

Topics: Adrenergic alpha-Antagonists; Anaphylaxis; Animals; Antigens; Drug Hypersensitivity; Female; Guinea Pigs; Immunoglobulin E; Male; Mice; Mice, Inbred Strains; Ovalbumin; Passive Cutaneous Anaphylaxis; Prazosin; Rats; Serum Albumin, Bovine; Species Specificity

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

The ability of 5 dissimilar monoisocyanates conjugated to ovalbumin (OA) as a carrier protein to induce pulmonary hypersensitivity towards the hapten specific component was assessed by using a previously described method based on the determination of a respiratory index (RI) in the guinea pig. The test chemicals included the commercially available p-tolyl and hexyl monoisocyanates (TMI and HMI), with 4-isocyanoto-4'-diphenylmethane (IDM), 4-isocyanoato-4'-methyldiphenylmethane (IMDM) and 1-isocyanato-methyl-1,3,3-trimethylcyclohexane (IMTC) as synthetized monoisocyanates. Guinea pigs were exposed daily to an aerosol of the OA conjugate of each monoisocyanate up to day 15. Increases in respiratory rate and/or respiratory collapse occurred in the guinea pigs exposed to TMI-OA and HMI-OA conjugates by days 9 and 15, with RI values of 155 and 177, respectively, being recorded. The greatest mean RI values in guinea pigs exposed to IDM-OA, IMDM-OA and IMTC-OA conjugates up to day 15 were 20, 25 and 22, respectively, and were not indicative of any pulmonary reaction. Guinea pigs exposed in parallel to each test conjugate did not exhibit any pulmonary reaction when they were exposed to OA on the challenge days. All these findings evidence pulmonary hypersensitivity as the result of exposure to TMI-OA and HMI-OA conjugates and suggest a high degree of conjugation and strong linkage of all the monoisocyanates with OA.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Aerosols; Animals; Cyanates; Drug Hypersensitivity; Female; Guinea Pigs; Lung; Ovalbumin; Respiration; Structure-Activity Relationship; Time Factors

An experimental model to investigate orbital granuloma formation in inbred rats was established. Animals sensitized to trinitrophenyl ovalbumin (TNP-OA) and challenged retro-orbitally with TNP-OA covalently linked o Sepharose 4B beads specifically developed a granulomatous response. This granulomatous reactivity was passively transferred into normal animals by lymph node cells, but not by serum antibody from sensitized donors. Lymphocytes which transfer granuloma formation in normal recipients were characterized by cell fractionation and membrane marker analysis. These experiments show that the effector cells capable of transferring granulomatous hypersensitivity are enriched in the lower density fractions on discontinuous Percoll gradients. These cells are lymphoblasts and express the W3/25 helper T lymphocyte marker. It was also demonstrated that lymphoid cells from sensitized donors in the higher density Percoll fraction appear to be incapable of adoptively transferring granulomatous responsiveness directly to normal recipients. However, incubation of these high density lymphocytes with specific antigen resulted in marked enhancement of their ability to transfer the disease. Antigen-induced activation also resulted in an increase in both lymphoblasts and the W3/25 marker. The authors conclude, therefore, that a subset of T cells which are lymphoblasts and express the helper-cell marker is responsible for granuloma formation in sensitized animals and is capable of transferring orbital granuloma formation to non-sensitized normal recipients.

Topics: Animals; Cell Count; Drug Hypersensitivity; Female; Granuloma; Intradermal Tests; Lymphocyte Activation; Orbital Diseases; Ovalbumin; Rats; Rats, Inbred Lew; T-Lymphocytes

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

In awake sensitized ponies, we studied the effect of aerosol ovalbumin challenge on ventilation, pulmonary mechanics, lung volume, and gas exchange before and after vagal blockade. We also challenged the left lung and measured respiratory rate (f) and right and left respiratory system resistance (RrsR, RrsL) before and after both left and bilateral vagal section. Bilateral ovalbumin aerosol challenge increased f, minute ventilation (VE), total respiratory system resistance (Rrs), and minimal volume, decreased dynamic compliance, total lung capacity, and arterial oxygen tension, and was without effect on tidal volume (VT), functional residual capacity, quasi-static lung compliance, and arterial carbon dioxide tension. Vagal blockade reversed the increase in f, VE, and Rrs and increased VT. Challenge of the left lung increased f and RrsL but did not alter RrsR. Bilateral vagal section reversed the tachypnea but unilateral section did not. Histopathologic lesions included acute fibrinopurulent obstructive bronchiolitis, bronchitis, edema, and alveolar distension. We conclude that local mechanisms are of critical importance in the pathogenesis of ovalbumin-induced airway obstruction in ponies, that increased sensitivity of airway smooth muscle to normal vagal tone may also play a role, and that tachypnea following challenge is caused by activity of pulmonary receptors with vagal afferent fibers.

Topics: Aerosols; Animals; Bronchi; Drug Hypersensitivity; Horses; Lung; Ovalbumin; Respiratory Hypersensitivity; Vagus Nerve

Topics: Animals; Antibody Formation; Carrier Proteins; Cyanates; Drug Hypersensitivity; Female; Hypersensitivity, Immediate; Immunization; Injections, Intraperitoneal; Injections, Intravenous; Male; Mice; Mice, Inbred Strains; Models, Biological; Ovalbumin; Passive Cutaneous Anaphylaxis; Reagins; Serum Albumin; Toluene 2,4-Diisocyanate

Reaginic antibodies to the benzylpenicilloyl determinant (BPO) and ovalbumin (OA) were induced readily in B6D2F1 mice by a single i.p. injection of either 1 or 10 mug of BPO4-OA suspended with 1 mg of Al(OH)3 in 0.5 ml of saline. Administration of conjugates consisting of the hapten coupled to the isologous, nonimmunogenic murine gamma-globulins (MgammaG), i.e., BPO9-MgammaG, BPO11-MgammaG, or BPO12-MgammaG, resulted in complete and specific suppression of the induction of the anti-BPO reaginic antibody response without affecting, however, the level of reaginic antibodies to OA. Further study of the effect of epitope density on the immunologic properties of BPOX-MgammaG revealed that a) the lightly haptenated conjugated, BPO1-MgammaG and BPO2.9-MgammaG, were not immunosuppressive, b) the conjugates, BPO4.3-MgammaG and BPO19-MgammaG, were partially tolerogenic, and c) the heavily haptenated conjugate, BPO31-MgammaG, was nontolerogenic. Moreover, most importantly, the ongoing anti-BPO response in sensitized mice was readily abrogated by either four daily or four weekly injections of BPO9-MgammaG. The immunosuppressive effect of BPO12-MgammaG conjugates was dose dependent, complete suppression being achieved with 200 mug of the tolerogen. The unresponsiveness to BPO of spleen cells from immunosuppressed donors was also maintained in adoptive cell transfer experiments in spite of the additional administration of the immunizing antigen under conditions expected to yield a secondary IgE response. Hence, it is suggested that, with special precautions to prevent unleashing an anaphylactic shock, treatment of penicillin-sensitive individuals with polyvalent conjugates of an appropriate number of BPO groups per human gamma-globulin molecule would constitute a rational immunotherapeutic procedure for the abrogation of the allergic response to BPO.

Topics: Animals; Antibody Formation; Antibody Specificity; Carrier Proteins; Drug Hypersensitivity; gamma-Globulins; Haptens; Immune Tolerance; Immunization, Passive; Immunoglobulin E; Immunosuppression Therapy; Immunotherapy; Mice; Mice, Inbred Strains; Ovalbumin; Penicillin G; Spleen; Structure-Activity Relationship

Cattle were more readily sensitised than guinea pigs to carboxymethylcellulose. Freund's complete adjuvant enhanced, but was not essential for, sensitisation. Schultz-Dale responses were obtained from pulmonary tissues of sensitised cattle but their sera failed to induce passive cutaneous anaphylaxis. Cattle could possibly be sensitised by carboxymethylcellulose contained in drug formulations.

Topics: Anaphylaxis; Animals; Antigens; Bordetella pertussis; Carboxymethylcellulose Sodium; Cattle; Cattle Diseases; Drug Hypersensitivity; Freund's Adjuvant; Guinea Pigs; Histamine Release; Injections, Intradermal; Injections, Intramuscular; Injections, Intraperitoneal; Injections, Intravenous; Methylcellulose; Muscle Contraction; Ovalbumin; Passive Cutaneous Anaphylaxis

Topics: Acid Phosphatase; Animals; Antibodies; Antigen-Antibody Complex; Cytoplasmic Granules; Drug Hypersensitivity; Erythrocytes; Fibroblasts; Freund's Adjuvant; gamma-Globulins; Histocytochemistry; Hypersensitivity, Delayed; Lymphocytes; Macrophages; Ovalbumin; Peroxidases; Plants; Rabbits; Rats; Sheep; Skin; Staining and Labeling; Time Factors

Topics: Adrenal Glands; Age Factors; Anaphylaxis; Animals; Bordetella pertussis; Drug Hypersensitivity; Female; Histamine; Hot Temperature; Humans; Hydrogen-Ion Concentration; Immunization; Injections, Intraperitoneal; Injections, Intravenous; Injections, Subcutaneous; Leukocyte Count; Mice; Mice, Inbred BALB C; Mice, Inbred CBA; Ovalbumin; Propionibacterium acnes; Serotonin

Topics: Air; Anaphylaxis; Animals; Aorta; Arachis; Blood Pressure; Dextrans; Drug Hypersensitivity; Emulsions; Female; Guinea Pigs; Histamine Release; Hypersensitivity; Iron-Dextran Complex; Lung; Male; Muscle Contraction; Oils; Ovalbumin; Perfusion; Polystyrenes; Pressure; Prostaglandins; Pulmonary Artery; Rabbits; Rats; Stimulation, Chemical; Suspensions; Time Factors

Topics: Adult; Anaphylaxis; Angioedema; Anhydrides; Animals; Antibodies; Antibody Formation; Antibody Specificity; Antigens; Aspirin; Carrier Proteins; Cattle; Drug Contamination; Drug Eruptions; Drug Hypersensitivity; Erythrocytes; Female; gamma-Globulins; Guinea Pigs; Haptens; Hemagglutination Tests; Humans; Hypersensitivity, Immediate; Immunodiffusion; Lysine; Male; Middle Aged; Ovalbumin; Passive Cutaneous Anaphylaxis; Serum Albumin; Skin Tests; Tannins; Urticaria

Topics: Animals; Antibodies; Aspirin; Cattle; Chromatography, Gel; Coliphages; Coombs Test; Drug Hypersensitivity; Glycosaminoglycans; Goats; Haptens; Hemagglutination Tests; Humans; Immune Sera; Neutralization Tests; Ovalbumin; Polystyrenes; Rabbits; Serum Albumin, Bovine

Topics: Acetylcholine; Animals; Blood Pressure; Blood Vessels; Bronchi; Bronchial Spasm; Drug Hypersensitivity; Extrachromosomal Inheritance; Female; Guinea Pigs; Histamine; Ileum; Male; Ovalbumin; Pilocarpine; Respiratory Hypersensitivity; Salivary Glands; Salivation; Serotonin; Skin

Topics: Adrenal Cortex Hormones; Aerosols; Anaphylaxis; Animals; Anti-Inflammatory Agents; Antigens; Benzyl Compounds; Betamethasone; Bronchi; Cortisone; Dexamethasone; Drug Hypersensitivity; Female; Fludrocortisone; Guinea Pigs; Hydrocortisone; Methylamines; Ovalbumin; Paramethasone; Prednisolone; Pyridines; Time Factors; Triamcinolone

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: Aerosols; Alpha-Globulins; Animals; Antigen-Antibody Reactions; Blood Group Antigens; Desensitization, Immunologic; Drug Hypersensitivity; gamma-Globulins; Guinea Pigs; Immunity, Maternally-Acquired; Immunization, Passive; Injections, Intraperitoneal; Ovalbumin

Topics: Animals; Arthus Reaction; Body Temperature; Drug Hypersensitivity; Guinea Pigs; Hypothermia, Induced; Immune Sera; Injections, Intradermal; Injections, Intravenous; Mice; Nitrobenzenes; Ovalbumin; Passive Cutaneous Anaphylaxis; Rabbits; Rectum

Topics: Acetylcholine; Animals; Bronchi; Drug Hypersensitivity; Female; Guinea Pigs; Histamine; Inbreeding; Male; Ovalbumin; Time Factors; Vagus Nerve

Topics: Absorption; Animals; Antibody Formation; Antigens; Aspirin; Cattle; Drug Hypersensitivity; Freund's Adjuvant; gamma-Globulins; Haptens; Hemagglutination Tests; Immune Sera; Immunodiffusion; Ovalbumin; Rabbits

Topics: Agglutination; Anaphylaxis; Animals; Antigen-Antibody Reactions; Blood Circulation; Blood Pressure; Blood Transfusion; Catheterization; Drug Hypersensitivity; Heparin; Hypotension; Leukocytes; Ovalbumin; Perfusion; Rats; Vasodilator Agents

Topics: Anaphylaxis; Animals; Antigen-Antibody Reactions; Antigens; Biomedical Research; Drug Hypersensitivity; Guinea Pigs; Hemagglutination; Ovalbumin; Penicillin G; Penicillins; Rabbits; Research; Serum Albumin; Skin Tests; Toxicology