Indoor allergen levels in settled airborne dust are higher in day-care centers than at home.

BACKGROUND: Early-life sensitization to indoor allergens predicts asthma development. The aim of this study was to compare allergen concentrations in day-care centers (DCC) with those in private homes. METHODS: Settled airborne dust was collected 4 times a year from 20 German DCC (620 samples) and from the homes of children and day-care workers (602 samples) using electrostatic dust collectors (EDC). The samples were analyzed with fluorescence enzyme immunoassays recognizing domestic mite allergens (DM), Fel d 1, Can f 1, and Mus m 1. Pet allergen thresholds that discriminate samples from homes with cats or dogs from those without were calculated using receiver-operating characteristics. Influences on allergen levels were analyzed using multilevel models. RESULTS: Allergen loads were on average higher in DCC than in homes. In DCC, 96% of the samples were positive for DM, 95% for Can f 1, 90% for Fel d 1, and 83% for Mus m 1. In homes, 84% contained DM, 48.5% Can f 1, 33% Fel d 1, and 43% Mus m 1. The threshold level for homes with dogs was 75 ng/m² Can f 1 (96.8% sensitivity, 96% specificity), and the threshold level for homes with cats was 46 ng/m² Fel d 1 (92% sensitivity, 94.9% specificity). In DCC, Can f 1 and Fel d 1 loads were higher than these thresholds in 37% and 54% of the samples, respectively. Allergen levels were significantly influenced by the season and room type; however, carpets on floors had no influence. CONCLUSIONS: Mite, mouse, cat, and dog allergens were mostly higher in DCC than in homes. Exposure to dog and cat allergens in DCC often reached levels of households with pets.

Quantification of protein and latex allergen content of various natural rubber latex products.

INTRODUCTION: The use of natural rubber latex (NRL) products can cause IgE-mediated allergic reactions in exposed people. The aim of this study was to quantify the content of protein and latex allergens of currently available NRL products to estimate the allergenic potential of these products. METHODS: 14 household articles (pacifiers, baby bottle nipples, condoms, household and disposable gloves, toy balloons, and Band-Aids) as well as 18 NRL examination gloves currently used by healthcare workers were investigated. Extracts of the examination gloves were prepared according to the standard method DIN EN 455-3, which contains requirements and testing for biological evaluation of single use medical gloves. The protein content was determined with a modified Lowry method. Latex allergen content was measured using an IgE-inhibition immunoassay with a mix of serum-sensitized patients as detection antibody sources and the latex ImmunoCAP as solid phase. The allergens Hev b 1, 3, 5, and 6.02 were determined using available immunoassays. RESULTS: In 5 out of 18 examination gloves, the protein content was under the detection limit. The other 13 gloves contained protein between 7.1 and 92.3 µg protein/g material. Five glove brands contained protein concentrations above the recommended reference value of 30 µg protein/g material. Latex allergen could be measured in 12 out of 18 NRL gloves. In only 3 gloves could none of the allergens Hev b 1, 3, 5, and 6.02 be detected. Protein and Hev b 1 could be measured in the examined childcare products, while the concentrations of the latex allergens Hev b 3, 5, and 6.02 were mostly under the detection limit. Boiling of childcare products led to a reduction of protein and allergen content. In some of the other daily-used NRL articles, the protein and allergen contents were even higher than in gloves. CONCLUSION: Our study demonstrated that protein, and particularly latex allergens, were detectable in currently available examination gloves as well as in household articles whereby a risk for sensitization and/or induction of allergic symptoms could not be excluded.

Identification of wheat flour allergens by means of 2-dimensional immunoblotting.

BACKGROUND: Wheat flour proteins are allergens for 60% to 70% of bakers with workplace-related respiratory symptoms. OBJECTIVE: The aim of the study was to investigate the variability of IgE antibody patterns of wheat flour-sensitized bakers and to identify the most frequently recognized allergens. METHODS: Water/salt-soluble wheat flour proteins from the cultivar Bussard were separated by using 2-dimensional gel electrophoresis with immobilized pH gradients. IgE-reactive proteins were identified by means of immunoblotting with sera of 10 subjects with baker's asthma. Mass spectrometric fingerprinting was used to identify the proteins most frequently recognized by IgE. RESULTS: The IgE immunoblots obtained with 10 different sera exhibited a remarkable heterogeneity. Each patient showed an individual IgE-binding pattern with 4 to 50 different allergen spots. Altogether, more than 100 IgE-binding protein spots were detected. Nine of the predominant IgE-binding protein spots were identified by using mass spectrometric fingerprinting. The obtained masses matched 2 different isoforms of glycerinaldehyde-3-phosphate dehydrogenase from Hordeum vulgare, triosephosphate isomerase from H vulgare, and serpin, a serine proteinase inhibitor from Triticum aestivum. CONCLUSIONS: The results show a great interindividual variation of IgE-binding patterns of wheat flour proteins in baker's asthma. The clinical relevance of the identified 4 new allergens will be further investigated in the near future.

Identification and characterization of cross-reactive natural rubber latex and Ficus benjamina allergens.

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.

Class I endochitinase containing a hevein domain is the causative allergen in latex-associated avocado allergy.

BACKGROUND: In the medical literature immunoglobulin (Ig)E-mediated sensitization to avocado is rarely reported. On the other hand, more than 50% of subjects having IgE-mediated natural rubber latex allergy are sensitized to avocado fruit as demonstrated by skin-prick testing and/or specific IgE measurements and about 10-20% report hypersensitivity reactions after ingesting avocado. OBJECTIVE: The underlying pathomechanism of latex-associated avocado allergy is still unknown. The conserved hevein domain of the major latex allergen prohevein (Hev b 6.01) is a ubiquitous chitin-binding protein structure that can be found in several plant proteins and may be responsible for the observed cross-reactivity between latex and avocado fruit. METHODS: Chitin-binding avocado proteins (CBAPs) were isolated by affinity-chromatography and their IgE-binding characteristics were studied by immunoblotting using the sera from 15 avocado-sensitized latex patients. Inhibition experiments using isolated hevein and CBAPs as inhibitor solutions were performed to study the immunological cross-reactivity between both protein species and to assess the role of the CBAPs as mediators in latex-associated avocado allergy. RESULTS: In 80% of avocado-sensitized subjects (n = 15), IgE antibodies directed against a 31-kDa allergen were detected by immunoblotting. This IgE-binding protein was identified by protein sequencing to be a class I endochitinase containing a hevein domain at the N-terminus. Purified native and digested (using simulated gastric fluid) endochitinase were able to completely block all avocado-specific IgE antibodies in six out of seven avocado patients. CONCLUSIONS: Sensitization to endochitinase class I containing a hevein domain is the main underlying pathomechanism in latex-mediated avocado allergy.

Differentiation between cosensitization and cross-reactivity in wheat flour and grass pollen-sensitized subjects.

BACKGROUND: To diagnose baker's asthma, occupational sensitization to wheat flour should be distinguished clearly from influences of cosensitization such as it may exist in pollinosis patients. To define the route of sensitization, the cross-reactivity between wheat flour and grass pollen allergens was investigated. METHODS: Two groups of atopic individuals with or without professional contact to wheat flour were screened by skin prick test for their sensitization to wheat flour and grass pollen and, in the case of hints for cosensitization, by Enzyme Allergo Sorbent Test (EAST). Cross-reactivity between wheat flour and grass pollen allergens was investigated by IgE binding inhibition assay using sera of 20 cosensitized individuals and by immunoblots. RESULTS: The immunological cross-inhibition between wheat flour and grass pollen proves some proteins to share common allergenic determinants. Significant differences between bakers and individuals not occupationally exposed to flour could be seen in the inhibition of IgE binding to wheat flour. While the IgE binding to wheat flour allergens was only slightly inhibited by grass pollen proteins in baker's sera its inhibition was nearly complete by this extract in sera of nonexposed atopics. Immunoblots indicate that wheat proteins with a molecular weight of about 8-12 and 22 kD preferentially react with sensitized bakers' IgE and not with IgE of other individuals. IgE binding to grass pollen allergens in immunoblots and inhibition experiments showed no differences between bakers and other subjects. CONCLUSION: In the group of atopics without professional contact to flour, the positive test results from wheat flour obviously ensue from contact to cross-reacting grass pollen proteins whereas bakers are exposed and sensitized to allergens of both sources. Such cross-inhibition experiments help to identify the source of sensitization which may be difficult to obtain in case of cross-reacting allergens.

EAST and CAP specificity for the evaluation of IgE and IgG antibodies to diisocyanate-HSA conjugates.

Sera of 54 symptomatic workers showing sensitization to isocyanate-human serum albumin (HSA) conjugates were subjected to parallel enzyme allergosorbent test (EAST) and CAP measurements to determine IgE antibodies to diphenyl-methane diisocyanate-HSA, toluene diisocyanate-HSA and hexamethylene diisocyanate-HSA. Results of both methods correlated rather well with each other. In comparison to the EAST results, the CAP values were twice as high, and 4, 17 and 13% more frequently positive findings were obtained with the three different antigens. Autoinhibition performed with both methods proved the specificity of IgE binding in 92% of sera in EAST and in 89% of sera in CAP if values of > or = 0.35 kU/l were considered. The total IgE level in sera influenced the antibody results. Four of 20 sera studied by autoinhibition had a total IgE of > 700 kU/l, and two of them did not show significant autoinhibition with all conjugates by CAP and one serum by EAST. In addition, three of these sera showed an elevated binding to control HSA only (0.31-0.5 kU/l), and two revealed only a slightly increased IgE binding when compared with the HSA control (ratio of isocyanate-HSA to HSA, < 2). Only 2 of the 16 sera with a total IgE level of < 700 kU/l yielded a noninhibitory positive CAP result, whereas all positive EAST values of these sera could be significantly blocked by autoinhibition. Therefore, we suggest regarding EAST and CAP IgE results to isocyanate-HSA as positive if they exceed HSA control by 100% and are above 0.35 kU/l. Weak positive CAP results (< or = kU/l), especially of sera with total IgE > 700 kU/l, should by confirmed by inhibition experiments. Twelve of 40 symptomatic isocyanate workers exhibited borderline or weakly increased IgG values for diisocyanate-HSA conjugates in the CAP system and IgG-EAST. HSA tested in EAST as a reference showed nearly the same results as the isocyanate-HSA conjugates. In the 23 inhibition experiments, IgG-binding specificity was not confirmed. These findings imply IgG measurement to be of no diagnostic value in isocyanate-induced airway disorders.