ovalbumin and trimellitic-anhydride

With the increasing morbidity and mortality of asthma, asthma aggravated by environmental pollution has drawn more attention. This study investigated the exacerbating effects of trimellitic anhydride (TMA), a typical pollutant, in ovalbumin (OVA)-induced asthmatic mice and the gene and protein expressions of TRPA1, V1, V2 in lung tissue. Female BALB/c mice were respectively administered for 42 days as follow: sensitized and challenged with OVA, sensitized and challenged with TMA, sensitized with OVA and challenged with OVA plus TMA, as well as sensitized and challenged with OVA plus TMA. 24 h after the last challenge, the changes in airway resistance (RI) and lung dynamic compliance (Cdyn) were tested. The levels of the inflammatory cells in blood and bronchoalveolar lavage fluid (BALF) were determined. The gene and protein expressions of TRPA1, V1, V2 in lung tissue were examined, and levels of interleukin (IL)-4, -13, substance P (SP), prostaglandin D

Topics: Allergens; Animals; Asthma; Calcium Channels; Disease Models, Animal; Environmental Pollutants; Female; Gene Expression Regulation; Humans; Lung; Mice; Ovalbumin; Phthalic Anhydrides; TRPA1 Cation Channel; TRPV Cation Channels

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

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

Both trimellitic anhydride (TMA), a small molecular weight chemical, and ovalbumin (OVA), a reference protein allergen, cause asthma with eosinophilia. To test the hypothesis that different allergens elicit symptoms of asthma via different effector pathways, gene expression was compared in lungs of Balb/c mice sensitized with either TMA or OVA, followed by intratracheal challenge with TMA conjugated to mouse serum albumin (TMA-MSA) or OVA, respectively. Sensitized animals challenged with mouse serum albumin (MSA) alone were controls. Seventy-two hours after challenge, lung eosinophil peroxidase indicated that both allergens caused the same significant change in eosinophilia. Total RNA was isolated from lung lobes of 6-8 animals in each of four treatment groups and hybridized to Affymetrix U74Av2 GeneChips. False discovery rates (q-values) were calculated from an overall F test to identify candidate genes with differences in expression for the four groups. Using a q-value cutoff of 0.1, 853 probe sets had significantly different expression across the four treatment groups. Of these 853 probe sets, 376 genes had an Experimental/Control ratio of greater than 1.2 or less than 1/1.2 for either OVA- or TMA-treated animals, and 249 of the 376 genes were uniquely up- or down-regulated for OVA or TMA (i.e., differentially expressed with the allergen). qRT-PCR analysis of selected transcripts confirmed the gene expression analysis. Increases in both arginase transcript and enzyme activity were significantly greater in OVA-induced asthma compared to TMA-induced asthma. These data suggest that pathways of arginine metabolism and the importance of nitric oxide may differ in OVA- and TMA-induced asthma.

Topics: Allergens; Animals; Arginase; Asthma; Disease Models, Animal; Eosinophilia; Eosinophils; Female; Gene Expression; Intubation, Intratracheal; Mice; Mice, Inbred BALB C; Microchip Analytical Procedures; Ovalbumin; Peroxidase; Phthalic Anhydrides; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger

Feeding a soluble antigen to an animal is known to cause a state of unresponsiveness against this antigen. If this antigen is given together with another antigen during the sensitization procedure, impairment of the response to the new antigen can also be seen, a phenomenon referred to as bystander suppression. The induction of tolerance against ovalbumin (OvA) and the effect of bystander suppression on the response to the hapten trimellitic anhydride (TMA), a cause of occupational asthma, were studied in Brown-Norway rats. Rats were fed either OvA-containing pellets or standard diet for 16 days before sensitization with the mixture of TMA and OvA. The animals were followed for 6 weeks after sensitization. Animals made tolerant to OvA showed a significantly suppressed delayed-type hypersensitivity (DTH) reaction against both OvA and TMA compared with the nontolerized control group at 5 weeks after sensitization, implying bystander suppression. By contrast, immunoglobulin (Ig)E and IgG antibody levels were suppressed only against OvA, whereas anti-TMA antibody levels were not affected. Airway eosinophilia after a single aerosol challenge at 6 weeks after sensitization using TMA conjugated to rat serum albumin, correlated with IgE anti-TMA levels in the group made tolerant to OvA and was not affected by OvA ingestion. In conclusion, suppressive factors released in ovalbumin-tolerant rats when they are challenged with ovalbumin, can suppress the response to trimellitic anhydride and this suppression is more pronounced for T-helper1-type responses.

Topics: Airway Resistance; Allergens; Animals; Disease Models, Animal; Dose-Response Relationship, Immunologic; Eosinophils; Haptens; Hypersensitivity, Delayed; Immune Tolerance; Immunoglobulin E; Immunoglobulin G; Male; Occupational Diseases; Ovalbumin; Phthalic Anhydrides; Rats; Rats, Inbred BN; Reference Values; Respiratory Hypersensitivity

Sensitization of guinea pigs by intradermal injections of the occupational allergen trimellitic anhydride (TMA) in oily vehicle has been shown to be very reproducible. We studied the effect of intradermal sensitization with ovalbumin (OA) in oily vehicle on immune and airway responses in guinea pigs. We also compared airway responses to trimellitic anhydride or Dermatophagoides farinae (DF; mite) with those to OA in guinea pigs intradermally sensitized to respective allergens. Three to four weeks after sensitization, the animals were challenged with intratracheal instillation of these allergens. Intradermal injections with OA developed dose-dependently specific IgG1 antibodies to OA demonstrated by ELISA. In animals sensitized with different doses of OA in corn oil vehicle, a challenge with OA induced a reversely dose-dependent airflow obstruction and airway plasma exudation. In contrast, animals sensitized with OA in saline vehicle had dose-dependent airway responses to OA. Challenge with OA caused an immediate peak and subsequently persistent airflow obstruction, whereas this response to either TMA guinea pig serum albumin or Df was slowly progressive in animals sensitized to respective allergens. The animals sensitized to TMA or Df may show a different profile of airway responses following the challenge compared to OA. Intradermal sensitization may be a valuable method of sensitization for the development of an animal model of airway allergy to different types of allergens, including chemicals or mites.

Topics: Allergens; Animals; Antigens, Dermatophagoides; Enzyme-Linked Immunosorbent Assay; Glycoproteins; Guinea Pigs; Immunization; Immunoglobulin G; Injections, Subcutaneous; Male; Mites; Ovalbumin; Phthalic Anhydrides; Respiratory System

Topics: Allergens; Animals; Bronchial Provocation Tests; Female; Guinea Pigs; Immunoglobulin G; Injections, Intradermal; Isocyanates; Male; Ovalbumin; Phthalic Anhydrides; Respiratory Hypersensitivity

We established a model of asthma induced by trimellitic anhydride (TMA) in guinea pigs and assessed the role of sensitization in the development of their bronchial hyperresponsiveness, and relationship between bronchial responsiveness and bronchial inflammation. Fourteen guinea pigs (sensitized group) were administered 1 mg/0.5 ml of trimellity 36-bovine serum albumin intramuscularly and 0.5 ml of complete Freund adjuvant on Day 1 as the priming dose. Booster doses were repeated on Day 15. By Day 28, all of the sensitized animals showed a high passive hemagglutination titer against trimellityl 14-ovalbumin. On Day 29, they were challenged by an inhalation of TMA (150 mg/m3) for 30 min, and respiratory resistance (Rrs) was monitored by the oscillation method. In all sensitized animals, Rrs increased immediately upon challenge and returned to baseline within 6 h. The bronchial reactivity to acetylcholine (Ach), measured 6 h after TMA challenge in the sensitized animals, increased significantly (p < 0.01) compared with that measured 24 h before challenge; that measured 24 h later was not different from that before challenge. There was also a significant difference (p < 0.01) in the number of eosinophils in the lamina propria and the epithelium 6 and 24 h after the challenge inhalation in the sensitized group. The increased airway responsiveness to Ach in the sensitized animals was correlated with an increase in the number of eosinophils in the lamina propria and the epithelium. These observations suggest that humoral antibody and eosinophils are involved in the pathogenesis of TMA-induced asthma.

Topics: Acetylcholine; Administration, Inhalation; Animals; Antibody Formation; Asthma; Bronchi; Bronchial Hyperreactivity; Bronchial Provocation Tests; Disease Models, Animal; Guinea Pigs; Hemagglutination Tests; Immunization; Lung; Male; Ovalbumin; Passive Cutaneous Anaphylaxis; Phthalic Anhydrides; Sheep

Conscious Hartley guinea-pigs were sensitized with trimellitic anhydride (TMA) or trimeric hexamethylene diisocyanate biuret (Des-N) by repeated intradermal injections or by inhalation exposure. Immediate- and delayed-onset pulmonary reactions were recorded during/after challenges with the hapten or protein conjugate of the hapten, respectively. The positive control was sensitized and challenged with ovalbumin (OA) by inhalation. Homocytotropic antibodies of the IgG1 type were determined to correlate antibody titres and pulmonary responses. Immediate-onset pulmonary reactions were apparent in guinea-pigs sensitized by inhalation of TMA and OA. Animals sensitized intradermally with TMA demonstrated more vigorous immediate-onset reactions than animals sensitized by inhalation. The measurement of breathing parameters demonstrated that the ability to detect immediate-onset airway hyperreactivity was best when the breathing rate and tidal and minute volumes were measured. None of the measurements for delayed-onset reactions presented conclusive results, since sensitized as well as naive guinea-pigs demonstrated a delayed increase of breathing frequency. Antigen-specific IgG1 antibodies were several orders of magnitude higher in intradermally sensitized animals compared with animals sensitized by inhalation. Although Des-N-sensitized guinea-pigs experienced high IgG1-antibody titres, no response of pulmonary hypersensitivity could be elicited after hapten or conjugate challenge. In summary, it would appear that pulmonary hypersensitivity depends on factors other than IgG1 antibody titres. For the screening of chemicals, the dermal induction and inhalation challenge protocol was revealed to be more sensitive than the inhalation induction and inhalation challenge protocol.

Topics: Administration, Inhalation; Animals; Cyanates; Female; Guinea Pigs; Hypersensitivity, Delayed; Hypersensitivity, Immediate; Immunoglobulin G; Isocyanates; Lung; Ovalbumin; Phthalic Anhydrides; Respiratory Hypersensitivity

It was previously shown that administration of conjugates of trimellitic anhydride with polyvinyl alcohol (TM-PVA) could suppress the IgE anti-TM immune response of mice sensitized with a TM-ovalbumin conjugate. In the present study, the existence of TM-specific B cell tolerance was shown by cell transfer experiments in which splenic B cells from mice treated with TM-PVA failed to interact with either the helper T (Th) cells of carrier primed recipients or with Th cells derived from carrier-primed donors. In contrast to previous findings from this laboratory indicating that tolerogenic conjugates of PVA and the 2,4-dinitrophenyl group led to both B cell tolerance and activation of suppressor T (Ts) cells, no evidence was obtained for the induction of Ts cells by TM-PVA. Thus, the induction of demonstrable Ts cells by hapten-PVA conjugates may depend on some property conferred by the haptenic group.

Topics: Animals; B-Lymphocytes; Carrier Proteins; Female; Immune Tolerance; Immunization, Passive; Immunoglobulin E; Immunosuppressive Agents; Mice; Ovalbumin; Phthalic Acids; Phthalic Anhydrides; Polyvinyl Alcohol; Radiation Chimera; Rats; Spleen; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory

Topics: Animals; Antibody-Producing Cells; Dose-Response Relationship, Immunologic; Haptens; Immune Tolerance; Immunization; Immunoglobulin E; Mice; Mice, Inbred Strains; Ovalbumin; Phthalic Anhydrides; Polyvinyl Alcohol; Rats

The i.p. injection of 1 microgram of TM3-OA or TM9-OA with 1 mg of A1(OH)3 into B6D2F1 mice elicited the production of antibodies of the IgE and other classes to the trimellityl (TM) group and ovalbumin (OA). The induction of anti-TM antibodies belonging to the IgE and other immunoglobulin classes was specifically suppressed by the administration of tolerogenic conjugates prepared by coupling trimellitic anhydride (TMA) to the hydrophilic non-immunogenic polymer, polyvinyl alcohol (PVA), prior to immunization with TM-OA conjugates. More importantly, established anti-TM responses were also suppressed by these TM-PVA conjugates. By contrast, however, treatment with TM-PVA conjugates did not affect either the primary or the established anti-OA antibody response. The tolerogenic effects of the PVA conjugates were dose-dependent and appeared also to be dependent on the epitope density. Treatment with these conjugates also prevented immunized mice from showing any symptoms of systemic anaphylaxis on challenge with polyvalent TM-protein conjugates. These findings indicate that these conjugates may have the potential of useful therapeutic agents for the treatment of TMA-induced pulmonary hypersensitivity diseases.

Topics: Anaphylaxis; Animals; Antibody Formation; Dose-Response Relationship, Immunologic; Female; Immune Tolerance; Immunoglobulin E; Mice; Ovalbumin; Passive Cutaneous Anaphylaxis; Phthalic Acids; Phthalic Anhydrides; Polyvinyl Alcohol

The specificities of antibodies of different Ig classes against trimellityl (TM)-human serum albumin were examined by a radioimmunoassay inhibition technique. The antisera were from workers exposed to trimellitic anhydride who had 4 differing respiratory diseases. The studies demonstrated similarities between reactivities of antibodies of different Ig classes in these workers in that inhibition of antibody to TM-HSA required markedly less TM-HSA on a molar basis than TM-ovalbumin (OA) or sodium trimellitate (NaTM), the hapten for the trimellityl group. The results appears best explained by formation of new antigenic determinants on altered HSA molecules with the TM group forming a component of some of the new antigenic determinants. NaTM in high concentration could not completely inhibit the IgG antibodies of 2 sera suggesting either very low affinity for the hapten or that some of the antibodies might possibly be directed against new antigenic determinants formed from the reaction of TMA with HSA but that the TM group might not be a part of the antigenic specificity. The results of some studies also suggested that there were similar antigenic specificities on TM-HSA and TM-OA but of lesser concentration or lower affinity on the latter molecules. Passive transfer studies using a serum containing IgE antibodies against TM-HSA demonstrated that these human IgE antibodies will passively sensitize rhesus monkey skin. Neutralization of the cutaneous IgE antibodies occurred with TM-HSA but not with a great molar excess of NaTM in analogy with the in vitro studies.

Topics: Antibodies; Antibody Specificity; Binding Sites, Antibody; Binding, Competitive; Humans; Immunization, Passive; Immunoglobulin A; Immunoglobulin E; Immunoglobulin G; Immunoglobulins; Ovalbumin; Phthalic Acids; Phthalic Anhydrides; Radioimmunoassay; Serum Albumin; Tricarboxylic Acids