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Notes: A 29-year-old female bakery shop assistant was occupationally sensitized to flour allergens and Aspergillusα-amylase (Asp o 2). The latter represents a strongly allergenic component of routinely used baking additives. The patient had repeatedly responded to the consumption of white bread with rhinitis, conjunctivitis, and, occasionally, wheal and flare reactions. She underwent allergologic investigations including oral challenge tests with commercially available bread loaves. Elevated specific IgE antibodies against bread extracts, Asp o 2, and flour allergens were detectable in her serum. The provocation test with bread resulted in a running nose together with a strong increase in nasal resistance. All symptoms subsided about 3 h after the challenge. None of the above symptoms could be observed when bread free of Aspergillusα-amylase was administered. This outcome provides evidence of a clinically relevant persistent allergenicity to Asp o 2 in bread.

Notes: Recent studies have demonstrated that allergy to natural rubber latex (NRL) is associated with cross-reactivity to certain foods. The aim of this study was to investigate the prevalence of NRL sensitization and allergy in children with atopic dermatitis (n=74). We also examined cross-reactions between latex and foods, and compared the frequency of suspected latex cross-reacting fruits in children with and without NRL-specific immunoglobulin E (IgE). Twelve of the 74 atopic children studied (16.2%; 95% confidence interval (CI), 8.7–26.6%) had circulating IgE antibodies to latex. These NRL-sensitized children were older and they showed significantly higher total IgE values (p〈0.003) when compared with the group of children without NRL sensitization. Of the specific food IgE evaluations, 18.4% (93 out of 505) were positive, and 69.9% were observed in the group of children with latex-specific IgE, most frequently to potato, tomato, sweet pepper, and avocado. An isolated latex-specific IgE response without food-specific IgE was never observed. Exclusively in the latex-positive group, conformity with the report of allergic symptoms after ingestion of food and increased food-specific IgE was found. Twenty children without proven latex sensitization showed increased food-specific IgE, most frequently to potato, banana, and chestnut. Avocado-specific IgE was never determined in this patient group. No significant differences were detected concerning the sensitization to potato, banana, and kiwi between NRL-sensitized children and the group of 20 children without latex-specific IgE. The competitive CAP inhibition using sera from children with specific IgE to both latex and food showed different cross-reactivities between latex and the specific food. A close relationship existed between latex and avocado (median inhibition: 100%), whereas sensitization to latex and kiwi seemed to be independent in our study group (inhibition: 〈25%). In particular, for potato, cross-reactivity and co-sensitization existed. Our study demonstrated that children with atopic dermatitis are a high-risk group for latex sensitization. Increasing age, additional sensitization to ubiquitous inhaled allergens, and enhanced total serum IgE values seemed to be important variables for latex sensitization and further sensitization to the latex-associated foods. Cross-reactivity and, in some cases, co-sensitization to specific fruits and vegetables, were observed.

Notes: Background The rubber elongation factor in Hevea rubber (Hev b 1) is one of the most important latex allergen and is leading cause oflatex type 1 hypersensitivity in children with spina bifida.Objective The aim of this study was to define the allergenic and antigenic epitopes of Hev b 1.Methods The immunoglobulin- (Ig)E and IgG antibody binding sites on Hev b 1 allergen were delineated by enzyme linked immunosorbent assay (ELISA) using synthetic overlapping peptides covering the whole Hev b 1 sequence. In order to improve the binding capacity and specificity all peptides were biotinylated at the N-terminal end via a 6-aminohexanoic acid as spacer and then adsorbed to streptavidin pre-coated microtitre plates. Fine mapping to define the essential amino acid residues for the antibody binding was achieved by using overlapping peptides with one amino acid offset.Results It was demonstrated that the IgE epitopes were located in different regions of Hev b 1 including the C-terminal segment (121–137) and the segments with amino acid residues of 30–49 and 46–64. Two monoclonal antibodies (MoAbs) II2F3 and II4G9 raised against purified Hev b 1 recognized the C-terminal segment only. The results of epitope mapping with three rabbit antisera revealed that five positive peptides, including the epitope peptides 31–49, 46–64 and 121–137, were involved in the antibody-binding sites. Eine mapping on the segments 46–64 and 121–137 showed that the two MoAbs reacted with the peptide 125–134 in the C-terminal region, whereas the peptide with amino acids 124–134 was essential for recognition by human IgE antibodies. Epitopes to rabbit polyclonal IgG and human IgE were also found to be involved in the amino acid residues of 47–59.Conclusion Our results indicate that the most allergenic/antigenic portions of Hev b 1 allergen are the C-terminal region and the region with amino acid residues of 31–64. In both regions, the minimal IgE-binding epitope is almost identical with the IgG-binding epitope.

Notes: Chironomids (non-biting midges) are known to cause IgE-mediated hypersensitivity in man. This study compares the cross-reactivity between the chironomid midge Cladotanytarsus lewisi (lgreen nimitti), a widespread cause of allergy in the Sudan and Chironomus riparius (=thummi, CTT) where larvae are used as pet fish food and where haemoglobins were previously shown to be major allergens. As with C. riparius, immature forms of C. lewisi also contain allergenic material since skin test responses to larval, pupal and adult extracts were obtained in Sudanese individuals. Crossed radioimmunoelectrophoresis of the C. lewisi larval and pupal extracts indicate that they contain a higher proportion of the allergenic fractions than adults. Further evidence of common allergen determinants between C. lewisi and C. riparius were obtained by the demonstration of positive skin-prick tests, in Sudanese patients, to extracts of larval, adult and isolated haemoglobin extracts of C. riparius. Cross-reactivity between C. lewisi and C. riparius was also demonstrable by RAST inhibition studies. A dose-dependent inhibition was observed using both the C. lewisi adult midge RAST and the C. riparius haemoglobin RAST, the two respective antigens, and sera from individuals hypersensitive to either C. lewisi or C. riparius. Due to the immunological cross-reactivity found between these distantly related species, we conclude that chironomids should be seen as significant environmental and occupational allergens.

Notes: Spina bifida patients are at a high risk of developing latex allergy. Recently, we found a relationship between the IgE responsiveness to latex allergen hevein and human leucocyte antigen (HLA) alleles DRB1*04 (DR4) as well as DQB1*0302 (DQ8). This study was carried out to investigate the association between HLA class II alleles and the specific IgE response to latex allergen Hev b 1 in spina bifida patients.〈section xml:id="abs1-2"〉〈title type="main"〉MethodsBlood samples from 103 unrelated German spina bifida patients exposed to latex products and from 90 unsensitized controls were examined. Genomic DNA isolation followed by HLA-D-specifc polymerase chain reaction (PCR) amplification was used to perform HLA typing of allelic polymorphisms in exon 2 of DQB1 and of DRB1,3,4,5 with sequence-specific oligonucleotide probes (SSOPs).〈section xml:id="abs1-3"〉〈title type="main"〉ResultsFifty-one out of 103 spina bifida patients were found to have anti-latex IgE antibodies; 40 had also anti-Hev b 1 antibodies. Further, we observed that 80% of the Hev b 1 responders underwent five or more surgeries whereas 55% of the Hev b 1 non-responders and 75% of the latex-non-responders underwent less than five surgical interventions. From the latex-sensitized group 33% showed an elevated phenotype frequency of DRB1*0701 (DR7) when compared with unsensitized patients (12%, P = 0.0095, Pc = NS) and with controls (17%; P = 0.035, Pc = ns). Fifteen out of 40 Hev b 1 responders also exhibited an elevated DR7 frequency when compared with latex-sensitive but Hev b 1-negative patients (38% vs 18%, P = NS) or with unsensitized controls (38% vs 17%, P = 0.013, Pc = NS).〈section xml:id="abs1-4"〉〈title type="main"〉ConclusionsAlthough we found that the DRB1*0701 (DR7) phenotype frequency was elevated in SB patients with latex- as well as with Hev b 1-IgE responsiveness, the analyses of the other class II alleles clearly demonstrate that the HLA-D region does not play a major role in the pathogenetic way of sensitization to Hev b 1.

Notes: Background Sensitization to various flours and flour additives in the baking industry has been known for some time. However, most studies refer to allergens in their native state.Objective The aim of our study was to find out how high temperatures during the baking process influence enzymatic and allergenie activities of the common flour additive alpha-amylase (Asp o 2), a relevant allergen for bakers derived from Aspergillus oryzae.Methods In order to assess the allergenicity of Asp o 2 during the baking process, four to 11 correspondingly sensitized bakers were investigated by Enzyme Allergo-Sorbent Test (EAST) with native Asp o 2 and Asp o 2 heated to 80, 90, 95, 99.8 or 200 C. Eurthermore, the enzymatic activity was assayed in simulated baking processes at the above mentioned temperatures.Results Elevated temperatures resulted in a gradual loss of IgE antibodies’ recognition of Asp o 2 in two commercially available enzyme products. The enzymatic activity decreased more rapidly. Heating the enzyme to 200°C abolished both the enzymatic and the allergenie activity of the enzyme.Conclusion Based on these results, alpha-amylase in flour additives partially retains its allergenicity. This finding could be relevant for consumers.

Notes: Fourteen bakers suffering from workplace-related respiratory symptoms and sensitized to soybean were studied. Twelve of them were also allergic to wheat flour, 10 to rye flour, and five to α-amylase of Aspergillus oryzae (Asp o 2). IgE estimation by RAST strongly indicated that the trypsin inhibitor and lipoxidase are major allergens of soybean. Various allergenic components could be characterized by immunoblotting after two-dimensional electrophoresis. Our RAST and immunoblotting results show an interindividually different allergic response to inhalative soybean constituents, and that the trypsin inhibitor (20 kDa, pI˜4.5) is an important inhalative soybean allergen recognized by IgE antibodies in the sera of 86% of the examined sensitized bakers.

Notes: A total of 109 subjects reporting symptoms indicating type I hypersensitivity reactions to natural rubber latex (NRL) gloves was included in this study, and 66 of them had latex-specific IgE antibodies. They underwent provocation tests by wearing two types of NRL gloves with high (n=103) and low (n=75) allergen contents. The first glove type caused positive skin reactions in 30% of IgE-positive and in 3% of IgE-negative subjects. After application of a commercially available skin protection (barrier) cream, the frequencies of positive skin responses in wearing tests increased to 41% and 7%, respectively. The gloves with low allergen content did not cause hypersensitivity without skin-protection cream but induced responses in 5% of IgE-positive subjects when this cream was applied. Corresponding findings were obtained in intraindividual comparisons of test results, which were possible in 69 cases. Of all wearing-test responders. 92% had latex-specific IgE antibodies. Our results demonstrate that high allergen contents in latex gloves frequently elicite skin responses in NRL-sensitized subjects, and that skin-protection creams may favor the uptake of allergens from gloves, thus increasing allergic reactions. The stipulation of a legally binding threshold limit value (TLV) for allergens in NRL products is urgently needed. This TLV should not be set above 2 ng allergen/g rubber.