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Allergic reactions to wood dust allergens are rare, and only few in vitro diagnostic tools and information about relevant allergens are available. To differentiate between protein-based allergy and probably clinically silent glycogenic sensitization, it is helpful to characterize the relevant protein allergens and specify IgE binding. The current case report deals with the occupational softwood allergy of a carpenter exposed to different wood dusts. Skin tests and IgE tests against wood were performed with specifically tailored ImmunoCAPs and cross-reactive carbohydrate determinants. Potential allergens were identified by IgE blots and tandem mass spectrometry. The clinical relevance was verified by challenge tests. Specific IgE to softwood (spruce, pine and larch wood), beech wood, natural rubber latex (NRL) and horseradish peroxidase (HRP) were detected. Allergens in spruce wood, the dominant allergen source, were identified as peroxidases. Softwood were the strongest inhibitors. HRP reduced IgE binding to softwood to <50%, indicating predominantly proteinogenic epitopes, whereas IgE binding to NRL and beech wood was reduced to >50% by HRP, indicating predominantly glycogenic IgE epitopes. Skin and challenge tests underlined that softwoods were the source of sensitization. For the polysensitized patient, a clinically relevant softwood allergy was diagnosed, not only by challenge tests but also with specifically tailored in vitro tools., (Copyright © 2011 S. Karger AG, Basel.)

Background: Cow hair and dander are important inducers of occupational allergies in cattle-exposed farmers. To estimate allergen exposure in farming environments, a sensitive enzyme immunoassay was developed to measure cow hair allergens., Methods: A sandwich ELISA was developed using polyclonal rabbitantibodies against a mixture of hair extracts from different cattle breeds. To assess the specificity of the assay, extracts from other mammalian epithelia, mites, molds and grains were tested. To validate the new assay, cow hair allergens were measured in passive airborne dust samples from the stables and homes of farmers. Dust was collected with electrostatic dust fall collectors (EDCs)., Results: The sandwich ELISA was found to be very sensitive (detection limit: 0.1 ng/ml) and highly reproducible, demonstrating intra- and interassay coefficients of variation of 4 and 10%, respectively. The assay showed no reactivity with mites, molds and grains, but some cross-reactivity with other mammalian epithelia, with the strongest reaction with goat. Using EDCs for dust sampling, high concentrations of bovine allergens were measured in cow stables (4,760-559,400 μg/m²). In addition, bovine allergens were detected in all areas of cattle farmer dwellings. A large variation was found between individual samples (0.3-900 μg/m²) and significantly higher values were discovered in changing rooms., Conclusion: The ELISA developed for the detection of cow hair proteins is a useful tool for allergen quantification in occupational and home environments. Based on its low detection limit, this test is sensitive enough to detect allergens in passive airborne dust., (Copyright © 2011 S. Karger AG, Basel.)

Background: Occupational allergy to latex is generally reported from occupational groups such as health care workers; however, few reports derive from other occupational settings., Methods: Two male subjects working as loom tuners in a textile manufacturing plant developed severe allergic reactions during the cutting and weaving of elastic bands, initially not suspected to contain latex constituents. Clinical evaluation and lung function tests were supplemented by skin prick testing, specific IgE evaluation and basophil activation assays with extracted elastic bands., Results: Both workers presented with rhinitis, episodes of tight chest and itchy eyes. Initial spirometry was normal with no significant reversibility; however, a histamine challenge test was positive in one worker. Skin prick testing to a battery of common inhalant allergens was negative; however, raised IgE levels were detected to latex using ImmunoCAP. On further testing, the specific IgE response was directed mainly to the major latex allergens rHev b 5, rHev b 6.01, rHev b 6.02 and nHev b 13. Basophils of the two workers, but not the unaffected control subjects, were strongly activated by extracts of the elastic and the cutting dust material., Conclusions: Workers are at high risk of becoming sensitised to latex allergens when exposed to excessive dust produced by loom tuning machines. Latex sensitisation should therefore be considered in workers developing unexplained work-related allergic reactions (including asthma) associated with unlabelled materials in the textile industry.

Background: A pharmaceutical industry worker was exposed to dust of gum arabic in the tablet coating plant and complained of work-related shortness of breath, chest tightness, runny nose, itching and redness of the eyes. This case was investigated for allergy to gum arabic and compared with a control group. The aim of the study was to identify the IgE-binding components responsible for the work-related symptoms., Methods: Skin prick tests (SPTs)and specific IgE (sIgE) measurements with environmental and occupational allergens, spirometry and a specific bronchial challenge with gum arabic were performed. One hundred and nineteen control subjects underwent SPT with gum arabic and 43 controls were tested for sIgE. Crossreactivity between gum arabic and horse radish peroxidase was investigated by IgE CAP inhibition. A combined procedure of immunoblotting and periodate treatment was applied to identify the epitope nature of gum arabic., Results: Allergy to gum arabic was shown by SPT, presence of sIgE and a positive bronchial challenge with gum arabic. Sensitization to gum arabic was demonstrated by SPT or sIgE in 7 and 5 controls, respectively. The results of inhibition with horse radish peroxidase, immunoblotting and periodate treatment suggest that gum arabic sIgE of the patient and 1 SPT-positive control subject were directed to the polypeptide chains of gum arabic. In contrast, gum arabic sIgE of the other controls reacted to carbohydrate components., Conclusions: Sensitization to gum arabic carbohydrate structures occurs casually in atopic patients with pollen sensitization without obvious exposure to gum arabic. This study suggests that allergy to gum arabic is mediated preferentially by IgE antibodies directed to polypeptide chains of gum arabic., (Copyright (c) 2006 S. Karger AG, Basel.)

Background: Detection of allergen-specific IgE antibodies in patients' sera plays a key role for the diagnosis of IgE-mediated allergy. If no validated test system is available, diagnostic tools must be developed, usually by coupling or binding the allergens to a solid phase. Streptavidin ImmunoCAP is a new solid phase for binding of allergens which can be used in the Pharmacia CAP system., Objective: It was the aim of this study to assess the diagnostic validity of Streptavidin ImmunoCAP., Methods: Biotinylation and allergen concentration for binding to Streptavidin ImmunoCAP were optimized and IgE obtained with natural rubber latex, obeche wood, wheat and rye flour Streptavidin ImmunoCAP were compared with the results of ImmunoCAP and Enzyme Allergo-Sorbent Test (EAST) using sera from patients complaining of workplace-related respiratory symptoms., Results: While the relation of biotin-label and protein was critical (best results were obtained with a 5- fold molar excess), labelled protein for coupling to streptavidin ImmunoCAP was applicable in a wide concentration range. On average, IgE values with streptavidin ImmunoCAP were as high as with ImmunoCAP but considerably higher than values obtained by EAST., Conclusion: Streptavidin ImmunoCAP is a valuable tool for sensitive and specific measurement of IgE binding to new allergens superior to cellulose disk-based methods.

Background: Peripheral blood mononuclear cells (PBMC) of subjects allergic to the insect-derived allergen Chi t 1--9 are characterized by an allergen-induced pronounced proliferation and increased expression of activation markers (CD25, HLA-DR, CD23). T cell lines showed an elevated percentage of V beta 8a-positive cells following stimulation by Chi t 1--9., Objective: The aim of the present study was to investigate whether V beta 8a dominance plays an important role in PBMC short-term cultures (24 h) as well. The role of exogenous added cytokines, especially IL-4, has been determined., Methods: The T cell receptor repertoire was measured with 16 monoclonal antibodies to epitopes on the variable region of the beta chain by flow cytometry. Patients allergic to Chi t 1--9 were compared to nonallergic subjects as well as to subjects with other occupational allergies. In addition, cytokines were determined intracellulary by flow cytometry. Studies were performed with PBMC cultured for 24 h., Results: After cultivation for 24 h without or with different stimuli (cytokines, allergen, phytohaemagglutinin), changes in the T cell receptor profile and the cytokine profile were measurable compared to the baseline value (without cultivation). Stimulation with IL-4 revealed increased percentages of V beta 8a-expressing cells in Chi t 1--9-sensitized patients. This IL-4-induced V beta 8a increase did not occur in PBMC from the two control subject groups (non-allergic and allergic to other allergens than Chi t 1--9). CONCLUSION In conclusion, the dominance of certain T cell receptor types seems to arise due to the exposure to specific allergens and cytokine production. Some T cell receptors are often affected, for example V beta 8a, whereas others only show minor variations. V beta 8a expression obviously plays an important role in Chi t 1-9 allergy., (Copyright 2001 S. Karger AG, Basel)

Background: The baking additives xylanase and cellulase were described as baking additives causing baker's asthma. It is not known whether monosensitization to these enzymes may occur., Methods: We present a case report of a baker with work-related asthma evaluated by skin prick test (SPT), enzyme-linked immunosorbent assay (EAST), immunoblot, EAST and immunoblot inhibition, and specific bronchial challenge. Fungal xylanase and alpha-amylase were measured by two-site enzyme immunoassays in products used by the patient at work., Results: Allergy to xylanase and cellulase was demonstrated by SPT, EAST, immunoblot and specific bronchial challenge (for xylanase only). No sensitization to alpha-amylase could be demonstrated, but there was a weak flour allergy as documented by EAST and immunoblot and a positive occupational-type challenge with high concentrations of rye flour. Four baking additives contained measurable amounts of fungal alpha-amylase and xylanase, without a correlation between these enzymes., Conclusions: We conclude that occupational asthma due to the baking additives xylanase and cellulase may occur without concomitant sensitization to alpha-amylase and only weak sensitization to flour., (Copyright 2001 S. Karger AG, Basel)

Background: An association between allergy to Ficus benjamina and natural rubber latex (NRL) has been suspected based on clinical and immunological observations. The responsible cross-reactive allergens have not been identified yet. This study was undertaken to investigate the cross-reactivity between hevein (Hev b 6.02, 4.7 kD), a major allergen of NRL, and F. benjamina and identify its counterpart in F. benjamina., Methods: 89 serum samples from subjects allergic to NRL were used in the study. Skin prick tests were performed with highly purified hevein and sap extract of F. benjamina. Specific IgE antibodies to NRL, F. benjamina and Hev b 6.02 were determined by the Pharmacia CAP method. Cross-reactivity among these allergens was investigated by means of CAP and immunoblot inhibition experiments. Two-dimensional gel electrophoresis separation and protein microsequencing were performed to identify the cross-reactive allergens in F. benjamina., Results: 67 out of 89 (75%) sera showed elevated IgE to hevein. Specific IgE to Ficus were found in 22 (24.7%) sera, and with 1 exception, all these sera also had IgE to Hev b 6.02. Results of CAP inhibition assays using 11 sera showing IgE to both Hev b 6.02 and Ficus demonstrated that the IgE to Ficus could be completely inhibited by Hev b 6.02 in 6 of 11 sera. Immunoblots and immunoblot inhibition assays revealed that a protein of about 45 kD in F. benjamina is strongly recognized by serum IgE. In addition, the IgE-binding reactivity to this 45-kD protein could be completely inhibited by preincubation of the sera with Hev b 6.02. N-terminal protein sequencing of 14 amino acids indicated that this 45-kD protein has a hevein-like domain at the N-terminal region and may belong to the endochitinase family., Conclusion: Latex-allergic patients are at higher risk of becoming sensitized to Ficus. Hev b 6.02 in latex is a major cross-reactive allergen and its counterpart in F. benjamina is an acidic protein with a molecular weight of about 45 kD and a hevein-like N-terminal domain., (Copyright 2000 S. Karger AG, Basel)

Background: Natural rubber latex (NRL) products are complex mixtures consisting of different allergenic components. Among them, Hev b 1 belongs to the important and well-characterized ones. To quantify the relevant allergen Hev b 1 in NRL products, a two-site monoclonal antibody (mAb)-based assay was developed., Methods: Two Hev b 1-specific mAbs with different epitope recognition and ability to bind simultaneously to an Hev b 1 molecule were used in the study. Both mAbs (II4F9 and II4G9) were enriched by in vitro production in a modular minifermenter and affinity purified. Wells of micro-ELISA plates coated with captured mAb II4G9 were incubated with samples containing Hev b 1. Bound Hev b 1 was detected by a combination of biotinylated mAb II4F9 as detection antibody and peroxidase-labeled avidin., Results: The optimized sandwich ELISA was highly reproducible in the linear range of the standard curve and Hev b 1 concentrations ranging from 12.5 to 400 ng/100 microl could be detected. The assay was suitable for the detection of Hev b 1 concentrations in latex sap and latex products, e.g. gloves, with a detection limit of 1.25 microg of Hev b 1/g of rubber. In a preliminary study with five different brands of latex gloves, Hev b 1 concentrations were found to be in the range of 18-40 microg per gram of rubber material, corresponding to 2-4% of the total extractable protein content in latex glove extracts., Conclusions: A sensitive sandwich assay was developed to quantify the latex allergen Hev b 1. This assay can be used to standardize latex extracts with regard to the content of the major allergen Hev b 1., (Copyright 2000 S. Karger AG, Basel)