Immediate Hypersensitivity Research Articles

A Study of Comparative on the Classification Criteria and Results of Several Chemicals of GHS Between Korea and Japan

PP-29-092 Background/Aims: The objectives of this study were to determine the current status of the Globally Harmonized System of Classification and Labeling of Chemicals (GHS) in Korea and Japan. We have also emphasized the need for international harmony in standardizing the GHS classification and finding methods to improve the GHS. Methods: To determine the difference between the GHS of Korea and Japan. We compared the published GHS classification results of 200 chemicals. We reviewed the concordance rate of Korea and Japan's GHS classifications and analyzed the basis of discordance rate. Results: As a result of the comparison between GHS classification results of Korea and Japan, the overall concordance rate was 70.9%. The target organ toxicity resulted in the least concordance rate showing 27% for the single exposure and 40.0% for the repeated exposure while the respiratory hypersensitivity and the aspiration hazard resulted in the highest concordance rate indicating 95.5% each. The major cause of discordance showed 74.3% for the “difference of reference,” 15.6% for the “difference of building block approach,” and 10.1% for the “difference of data in the same reference.” Conclusion: As a result, chemical study of Korea and Japan, health and environmental hazard for the 200 chemicals classified discordance rate was 29.1%, all items matching the category, the results were just a matter of 5 chemicals. To minimize the confusion which may occur because of the introduction of GHS, the uniformity of GHS classification criteria, and results between countries os needed, and the effort for achieving the actual harmony of classification results of chemicals through the establishment of basis for the joint research like GHS Expert Committee between countries or multi parties is also demanded.

Further development of the NCTC 2544 IL-18 assay to identify in vitro contact allergens

Several European Union legislations request the use of in vitro methods for toxicological evaluations, including sensitization, in order to increase consumer safety but also to reduce the use of animals. The EU project SENS-IT-IV addresses the need of developing predictive in vitro tests to assess contact and respiratory hypersensitivity reactions. In this context, we have recently reported the possibility to use IL-18 production in the human keratinocyte cell line NCTC 2544 to discriminate contact sensitizer from irritants and low molecular weight respiratory allergens. The aims of the present study were to further develop this assay in order to optimize experimental conditions; to develop a 96-well plate format to establish a high throughput assay; to test the performance of other available keratinocyte cell lines, and to understand the signal transduction pathway involved in p-phenylenediamine (PPD)-induced IL-18 production.If cells reach confluence at the moment of treatment, the ability to identify contact allergens is lost; therefore a careful check for the optimal cell density using PPD as reference contact allergen is critical. In our hands, a cell density of 1–2.5×105cells/ml gave optimal stimulation. In order to develop a high throughput test, cells seeded in 96-well plate were exposed to contact allergens (2,4-dinitrochlorobenzene, p-phenylenediamine, isoeugenol, cinnamaldehyde, tetramethylthiuram disulfite, resorcinol, cinnamic alcohol and eugenol), irritants (phenol, sodium laurel sulphate, lactic acid and salicylic acid) and respiratory allergens (hexachloroplatinate, diphenylmethane diisocyanate, trimellitic anhydride). A selective increase in total (intracellular plus released) IL-18 was observed 24h later in cells treated with contact allergens, whereas no changes were observed following treatment with respiratory allergens and irritants, confirming previous results obtained in a 24-well format assay. A selective induction of IL-18 was also obtained testing with PPD other keratinocyte cell lines, namely HPKII and HaCaT, with the HPKII showing the highest stimulation index. Regarding the signal transduction pathway, we could demonstrate using selective inhibitors a role for oxidative stress, NF-κB and p38 MAPK activation in PPD-induced IL-18 production.In conclusion, results obtained suggest that the production of IL-18 represents a promising endpoint for the screening of potential contact allergens. The assay can be performed in a 96-well plate format, different keratinocyte cell lines can be used, and a role for oxidative stress in contact allergen-induced IL-18 was demonstrated.

Identification of covalent binding sites of ethyl 2-cyanoacrylate, methyl methacrylate and 2-hydroxyethyl methacrylate in human hemoglobin using LC/MS/MS techniques

Acrylates are used in vast quantities, for instance in paints, adhesive glues, molding. They are potent contact allergens and known to cause respiratory hypersensitivity and asthma. Here we study ethyl 2-cyanoacrylate (ECA), methyl methacrylate (MMA) and 2-hydroxyethyl methacrylate (HEMA). There are only limited possibilities to measure the exposure to acrylates, especially for biological monitoring. The aim of the present study was to investigate the chemical structures of adducts formed after reaction of hemoglobin (Hb) with ECA, MMA, and HEMA. This information may be used to identify adducted Hb peptides for biological monitoring of exposure to acrylates. Hb-conjugates with ECA, MMA, and HEMA were synthesized in vitro. The conjugates were digested by trypsin and pronase E. Adducted peptides were characterized and analyzed by liquid chromatography and nano electro spray/hybrid quadrupole time-of-flight mass spectrometry (MS) as well as tandem quadrupole MS. The search for the adducted peptides was facilitated by visualizing the MS data by different computer programs. The results showed that ECA binds covalently to cysteines at the 104 position in the α and the position 112 in the β-chains in Hb. MMA and HEMA bound to all the cysteines in both chains, Cys 104 in the α-chain and Cys 93 and 112 in the β-chain. The full-length spectra of in un-digested Hb confirmed this binding pattern. There was no reaction with N-acetyl- l-lysine at physiological pH. The adducted peptides were possible to measure using LC/MS/MS in selected reaction monitoring mode. These peptides may be used for biological monitoring of exposure to ECA, MMA and HEMA.

Inbred Chinese Wuzhishan (WZS) Minipig Model for Soybean Glycinin and β-Conglycinin Allergy

Validated murine models have been built to assess the potential allergenicity of novel proteins. Large animals, such as pigs, share more similarities to humans in physiology and immunology than murine. Among Chinese minipigs, Wuzhishan (WZS) minipigs have the highest inbreeding coefficient, more stable heredity, and less variability, which were gastrically sensitized and excitated with diets containing 4% glycinin or 4% beta-conglycinin or neither to induce anaphylactic reactions in the present study. In glycinin- and beta-conglycinin-sensitized animals, diarrhea symptoms and skin wheal and flare responses were observed. In comparison to the control, after oral excitation with glycinin or beta-conglycinin, the serum IgE was increased by 34.4 or 38.4% and the serum histamine was increased by 42.1 or 46.9%, respectively. In addition, the serum IFN-gamma were reduced by 12.4 or 30.0%, respectively. The jejunum histamine level of beta-conglycinin-sensitized animals was increased by 196.6%, while the number of mast cells in the submucosa of jejunum and ileum of the glycinin-sensitized animals declined by 48.1 and 45.0%. In conclusion, the WZS minipig allergy models induced by soybean glycinin and beta-conglycinin represent type-I hypersensitivity reactions mediated by IgE, which could potentially be useful in determining the potential allergenicity of novel proteins.

A method for measuring mouse respiratory allergic reaction to low-dose chemical exposure to allergens: An environmental chemical of uncertain allergenicity, a typical contact allergen and a non-sensitizing irritant

Our aim was to improve a method for detecting respiratory hypersensitivity by testing three confirmed respiratory allergens (trimellitic anhydride [TMA], phthalic anhydride [PA] and toluene diisocyanate [TDI]), an environmental chemical of uncertain allergenicity (2,4- d-butyl [DB]), a confirmed contact allergen (2,4-dinitrochlorobenzene [DNCB]) and a standard irritant (sodium dodecyl sulfate [SDS]). BALB/c mice were topically sensitized (nine times in 3 weeks) with these chemicals, then challenged via the trachea. One day post-challenge, samples were taken from the mice to assay for total immunoglobulin (IgE and IgG 1) levels in serum and bronchoalveolar lavage fluid (BALF); differential cell counts and cytokine/chemokine levels in BALF; lymphocyte counts and surface antigen expression on B-cells within lung-associated lymph nodes (LNs); ex situ cytokine production by cells from these LNs; and gene expression in BALF (CCR3) and LNs (CCR4, STAT6 and GATA-3). The three confirmed respiratory allergens and DB induced immune response characteristic of immediate-type respiratory reactions, as evidenced by increased total IgE and IgG 1 levels; influx of eosinophils, neutrophils, chemokines and cytokines in BALF; increased surface antigen expression on B-cells in LNs; increased Th2 cytokine production in LNs; and increased respiratory allergy-related gene expression in both BALF and LNs. In contrast, DNCB and SDS treatments yielded, at most, insignificant increases in all respiratory allergic parameters. Thus, the protocol was equally suitable for use with an environmental chemical of unknown allergenicity and for typical respiratory allergens. Since the protocol differentiated respiratory allergens from contact allergens and irritants, it may be useful for detecting respiratory allergy related to environmental chemicals.

Severe respiratory adverse events associated with rituximab infusion

Sirs, We read with great interest the article in Pediatric Nephrology entitled “Fatal pulmonary fibrosis after rituximab administration” by Chaumais et al. [1]. We believe that patients who have lung complications are at an extremely high risk of respiratory adverse events after rituximab infusion. Respiratory events, such as cough, bronchospasm, and dyspnea, are relatively common adverse effects of rituximab. At our center, 28 patients have received rituximab to date (21 with steroid-dependent nephrotic syndrome and 7 with steroid-resistant nephrotic syndrome). Ten (36%) of these patients have experienced respiratory events, such as an infusion reaction. Several case reports have described severe respiratory injuries associated with the use of rituximab, such as interstitial pneumonitis and acute respiratory distress syndrome, although the precise frequency is unknown. There have been 32 reports (total of 62 cases) of rituximabinduced severe respiratory adverse events [2]. In these reports, elderly people were considered to be at high risk, and most of the patients were older than 50 years (median age 62 years, range 9–88). Of these 62 patients (85%), 53 had lymphoma or B-cell lymphocytic leukemia, and 46 (74%) suffered from interstitial pneumonitis and other complications, including bronchiolitis obliterans with or without organizing pneumonia (4 patients), acute respiratory distress syndrome (3), organizing pneumonia (3), hypersensitivity pneumonia (2), and pulmonary fibrosis (1); for three patients, there was no definite diagnosis. Prompt initiation of steroid therapy appears to have conferred benefit, although some cases resulted in fatal outcomes (12 patients, 19%). Most of the patients reported were elderly, but there were two pediatric patients and, surprisingly, both of these children suffered from refractory nephrotic syndrome [1, 2]. Bitzan et al. reported a 14-year-old boy who suffered frequent relapsing nephrotic syndrome despite the use of various immunosuppressants [2]. Respiratory symptoms (cough, chest pain, fatigue, and fever) occurred 19 days after the last infusion of rituximab, and a chest computed tomography demonstrated the presence of diffuse, groundglass infiltrates in both lungs, which was compatible with interstitial pneumonitis. The patient's respiratory symptoms improved without recurrence. However, he was successfully treated with prednisolone and antibiotics. It may be interesting to investigate whether pediatric patients with nephrotic syndrome have a high risk of interstitial pneumonitis. At the present time, we recommend that a routine chest radiograph is obtained before riuximab infusion is initiated. We believe that clinicians should be highly alert to the possibility of severe respiratory adverse events and should never administer rituximab to patients already suffering from lung disease, such as pleural effusion, pneumonia, and atelectasis.

Type IV delayed-type hypersensitivity of the respiratory tract due to budesonide use: report of two cases and a literature review

Respiratory type-IV hypersensitivity reactions due to corticosteroids is a rare phenomenon. We describe two such cases. The first is a 37- year-old atopic woman who developed labial angioedema and nasal itching after the use of budesonide nasal spray. A month later, after the first puffs of a formoterol/budesonide spray prescribed for asthma, she noticed symptoms of tongue and oropharyngeal itching and redness with subsequent dysphagia, labial and tongue angioedema, and facial oedema. The second is a 15-year-old non-atopic woman who reported pruritic eruptions around the nostrils after using a budesonide nasal spray. A year later she presented with nasal pruritus with intense congestion and labial and facial oedema after using the same spray. Both patients were evaluated with patch-tests using the commercial T.R.U.E. test, a budesonide solution, and corticosteroid creams. Test evaluation was performed at 48 and 96 hours. In both patients, patch tests were positive to budesonide (++) on the second day. The first patient also had a positive (+) reaction to tixocortol-21-pivalate. All the other patch tests were negative. Clinicians should be aware that hypersensitivity reactions may occur during the use of nasal or inhaled corticosteroids.

Immediate hypersensitivity to carob pods

Immediate hypersensitivity to carob pods

Innate Immunity toAspergillusSpecies

All humans are continuously exposed to inhaled Aspergillus conidia, yet healthy hosts clear the organism without developing disease and without the development of antibody- or cell-mediated acquired immunity to this organism. This suggests that for most healthy humans, innate immunity is sufficient to clear the organism. A failure of these defenses results in a uniquely diverse set of illnesses caused by Aspergillus species, which includes diseases caused by the colonization of the respiratory tract, invasive infection, and hypersensitivity. A key concept in immune responses to Aspergillus species is that the susceptibilities of the host determine the morphological form, antigenic structure, and physical location of the fungus. In this review, we summarize the current literature on the multiple layers of innate defenses against Aspergillus species that dictate the outcome of this host-microbe interaction.

House dust mite major allergen Der f 1 enhances proinflammatory cytokine and chemokine gene expression in a cell line of canine epidermal keratinocytes

House dust mite (HDM) allergens are the most common allergens involved in the induction of IgE-mediated hypersensitivity. Recently, epicutaneous sensitization with HDM allergens has been emphasized in the development of atopic dermatitis (AD); however, direct stimulation of canine keratinocytes by mite allergens has not been well investigated. In the present study, we investigated the effects of Der f 1, a major allergen of Dermatophagoides farinae, on cytokine and chemokine gene expression in a canine keratinocyte cell line, CPEK. CPEK constitutively expressed mRNA for TNF-α, IL-12p35, IL-18, GM-CSF, TGF-β, IL-8/CXCL8, TARC/CCL17, CTACK/CCL27 and MEC/CCL28. Of all the cytokines and chemokines investigated in CPEK, transcription levels of GM-CSF, IL-8/CXCL8 and TNF-α mRNA were significantly enhanced by stimulation with Der f 1. The present results suggest that Der f 1 can directly augment inflammatory cytokine and chemokine production from keratinocytes, and may initiate allergic inflammation independently of Type-I hypersensitivity.

Respiratory and skin hypersensitivity reactions caused by a peptide coupling reagent

TBTU (2-[1H-benzotriazol-1-yl]-1,1,3,3-tetramethyluronium tetrafluoroborate; CAS No: 125700-67-6) and HBTU (2-[1H-benzotriazol-1-yl]-1,1,3,3-tetramethyluronium hexafluorophosphate; CAS No: 94790-37-1) are increasingly used as coupling reagents for the stepwise synthesis of peptides. There have been isolated reports...

Use of long term dermal sensitization followed by intratracheal challenge method to identify low-dose chemical-induced respiratory allergic responses in mice

The inhalation of many types of chemicals, including pesticides, perfumes, and other low-molecular weight chemicals, is a leading cause of allergic respiratory diseases. We attempted to develop a new test protocol to detect environmental chemical-related respiratory hypersensitivity at low and weakly immunogenic doses. We used long-term dermal sensitization followed by a low-dose intratracheal challenge to evaluate sensitization by the well-known respiratory sensitizers trimellitic anhydride (TMA) and toluene diisocyanate (TDI) and the contact sensitizer 2,4-dinitrochlorobenzene (DNCB). After topically sensitizing BALB/c mice (9 times in 3 weeks) and challenging them intratracheally with TMA, TDI, or DNCB, we assayed differential cell counts and chemokine levels in bronchoalveolar lavage fluid (BALF); lymphocyte counts, surface antigen expression of B cells, and local cytokine production in lung-associated lymph nodes (LNs); and antigen-specific IgE levels in serum and BALF. TMA induced marked increases in antigen-specific IgE levels in both serum and BALF, proliferation of eosinophils and chemokines (MCP-1, eotaxin, and MIP-1β) in BALF, and proliferation of Th2 cytokines (interleukin (IL)-4, IL-10, and IL-13) in restimulated LN cells. TDI induced marked increases in levels of cytokines (IL-4, IL-10, IL-13, and IFN-γ) produced by restimulated LN cells. In contrast, DNCB treatment yielded, at most, small, nonsignificant increases in all parameters. Our protocol thus detected respiratory allergic responses to low-molecular weight chemicals and may be useful for detecting environmental chemical-related respiratory allergy.

How irritating: the role of TRPA1 in sensing cigarette smoke and aerogenic oxidants in the airways

Airway irritants cause a variety of lung pathologies. Two separate studies, the first recently reported in the JCI by Bessac et al. and the second reported by Andrè et al. in the current issue of the JCI (see the related article beginning on page 2574), have identified irritants that activate transient receptor potential cation channel, subfamily A, member 1 (TRPA1) receptors in airway sensory neurons, resulting in neurogenic inflammation and respiratory hypersensitivity. The identification of TRPA1 activation by toxicants from cigarette smoke and polluted air, such as crotonaldehyde, acrolein, and oxidizing agents such as hydrogen peroxide, is an important finding. These two studies enhance our understanding of how pollution and cigarette smoke can damage airway function and will hopefully pave the way for the development of rational alternative therapeutics for such airway injury.

Immunology of Dermatophytosis

The immune response to infection by dermatophytes ranges from a non-specific host mechanism to a humoral and cell-mediated immune response. The currently accepted view is that a cell-mediated immune response is responsible for the control of dermatophytosis. Indeed, some individuals develop a chronic or recurrent infection mediated by the suppression of a cell-mediated immune response. The immune response to Trichophyton is unusual in that this fungus can elicit both immediate hypersensitivity (IH) and delayed-type hypersensitivity (DTH) in different individuals when they are submitted to a skin test reaction. Understanding the nature and function of the immune response to dermatophytes is an exciting challenge that might lead to novel approaches in the treatment and immunological prophylaxis of dermatophytosis.

Human Lung Epithelial Cells Express a Functional Cold-Sensing TRPM8 Variant

Several transient receptor potential (TRP) ion channels sense and respond to changes in ambient temperature. Chemical agonists of TRP channels, including menthol and capsaicin, also elicit sensations of temperature change. TRPM8 is a cold- and menthol-sensing ion channel that converts thermal and chemical stimuli into neuronal signals and sensations of cooling/cold. However, the expression and function of TRPM8 receptors in non-neuronal cells and tissues is a relatively unexplored area. Results presented here document the expression and function of a truncated TRPM8 variant in human bronchial epithelial cells. Expression of the TRPM8 variant was demonstrated by RT-PCR, cloning, and immunohistology. Receptor function was characterized using the prototypical TRPM8 agonist, menthol, and exposure of cells to reduced temperature (18 degrees C). The TRPM8 variant was expressed primarily within endoplasmic reticulum membranes of lung epithelial cells and its activation was attenuated by thapsigargin, the cell-permeable TRPM8 antagonist N-(4-tert-butylphenyl)-4-(3-chloropyridin-2-yl)piperazine-1-carboxamide, and shRNA-induced suppression of TRPM8 expression. Activation of the TRPM8 variant in lung cells was coupled with enhanced expression of the inflammatory cytokines IL-6 and IL-8. Collectively, our results suggest that this novel TRPM8 variant receptor may function as a modulator of respiratory physiology caused by cold air, and may partially explain asthmatic respiratory hypersensitivity to cold air.

Alcian Blue and Pyronine Y histochemical stains of paraffin embedded sections provide multi‐parameter morphologic assessment in experimental models of pulmonary disease

Pulmonary researchers often assess morphologic parameters to determine disease progression or response to novel therapeutics. Detection of morphologic parameters by special stains is essential for assessment. We show novel application of Alcian Blue and Pyronine Y histochemical stains in experimental models of lung disease to provide multi‐parameter tissue assessment. The Alcian Blue provided intrinsic counterstaining through pale blue staining of nuclei. Acidic mucins of airway epithelia or submucosal glands were stained deep blue (sialylated mucins), red (sulfated mucins), or purple (sialylated/sulfated mucins). Elastic fibers stained red within the walls of pulmonary arteries, connecting airways, alveolar septa and subpleural space. Mast cells were readily detected in select species by red granular cytoplasmic staining. Mucin alterations were readily detected in a respiratory syncytial virus hypersensitivity model and elastin loss in a model of bronchiectasis. Experimental models of respiratory disease which had representative staining included: inbred mice, rat, ferret, cat, dog, sheep and pig. This histochemical stain assesses multiple morphologic parameters often required by pulmonary investigators, is cost effective, simple to perform, and reduces the needs for repeated serial sectioning. This work is supported by the Department of Pathology (University of Iowa) and the NIH.

An investigation of respiratory hypersensitivity and work related allergens on pig slaughters in Taiwan

An investigation of respiratory hypersensitivity and work related allergens on pig slaughters in Taiwan

Th2 Cytokines in Skin Draining Lymph Nodes and Serum IgE Do Not Predict Airway Hypersensitivity to Intranasal Isocyanate Exposure in Mice

Isocyanate exposure in the workplace has been linked to asthma and allergic rhinitis. Recently, investigators have proposed that Th2 cytokine responses in lymph nodes draining the site of dermal application of chemicals including isocyanates may be used to identify sensitizers that cause asthma-like responses. The purpose of this study was to determine if the cytokine profile induced after dermal sensitization with isocyanates and serum IgE predict immediate (IHS) and methacholine-induced late (LHS) respiratory hypersensitivity responses after intranasal challenge. Dermal application of hexylmethane diisocyanate (HMDI), toluene diisocyanate (TDI), or methylene diisocyanate (MDI) significantly increased interleukin-4 (IL-4), IL-5, and IL-13 secretion in parotid lymph node cells. Isophorone diisocyanate (IPDI) increased IL-4 and IL-13, but not IL-5. Tolyl(mono)isocyanate (TMI), tetramethylene xylene diisocyanate (TMXDI), or the contact sensitizer dinitrochlorobenzene (DNCB), only induced minor increases in some of the Th2 cytokines. HMDI, TDI, MDI, and IPDI elicited greater increases in total serum IgE than DNCB, TMI, and TMXDI. All chemicals except TMXDI caused IHS after intranasal challenge of sensitized female BALB/c mice. Only HMDI-, TMI-, or TMXDI-sensitized and challenged mice had increases in LHS. All chemicals elicited epithelial cytotoxicity indicative of nasal airway irritation. The discordance between dermal cytokine profiles and respiratory responses suggests that dermal responses do not necessarily predict respiratory responses. Serum IgE also was not predictive of the respiratory responses to the isocyanates, suggesting that other unknown mechanisms may be involved.

Immediate hypersensitivity to paraphenylenediamine

Immediate hypersensitivity to paraphenylenediamine

RESEARCHES REGARDING THE DIAGNOSIS OF ATOPIC DERMATITIS IN DOG

Atopic dermatitis is a skin disease that occurs after a type I hypersensitivity reaction, reaction which appears in individuals that have a genetic predisposition to produce excessive quantities of immunoglobulin E to certain environmental allergens: house dust mites, pollens, fungi, epithelia, hair, wool, feathers