Search

BackgroundLimited is known on the profiles of apple allergy in China.ObjectiveTo explore the clinical significance of apple allergen components in northern China.MethodsThis study recruited 40 participants and categorized into apple tolerance (n = 19) and allergy (n = 21) group. The latter was categorized into oral allergy symptoms (OAS, n = 14) and generalized symptoms (GS, n = 7). All participants underwent ImmunoCAP screening to assess sIgE levels of birch, apple, and their components.ResultsThe sensitization rates were 90% for Bet v 1, 85% for Mal d 1, 35% for Bet v 2, and 20% for Mal d 3. The overall positive rate for apple allergens was 97.5%, with half demonstrating mono-sensitization to Mal d 1. Birch, Bet v 1 and Mal d 1 sIgE levels had consistent areas under the curve (AUC 0.747, p = 0.037; AUC 0.799, p = 0.012; AUC 0.902, p

Xiaoyan Wang,1,2 Lijia Chen,1 Jiaqi Ding,3 Hongtian Wang,1 Xueyan Wang1,2 1Department of Allergy, Beijing Shijitan Hospital, Capital Medical University, Beijing, 100038, People’s Republic of China; 2Beijing Laboratory of Allergic Diseases, Beijing Municipal Education Commission, Beijing, 100038, People’s Republic of China; 3Department of Otolaryngology, Qingdao Women and Children’s Hospital Affiliated to Qingdao University, Qingdao, Shandong, 0355729, People’s Republic of ChinaCorrespondence: Xueyan Wang, Hongtian Wang, Tieyi Road, No. 10th, Haidian District, Beijing, 100038, People’s Republic of China, Tel +86 13810570961 ; +86 13391836668, Email allergy_wxy@126.com; wht301@263.netPurpose: To investigate the major allergen components associated with birch pollen allergy in northern China and elucidate clinical relevance to pollen food allergy syndrome (PFAS).Methods: Fifty-eight patients were recruited for a cross-sectional study and categorized into two groups: PFAS group and non-PFAS group, as well as apple allergy group and non-apple allergy group. The sIgE levels of birch pollen and its components, namely Bet v 1, Bet v 2, Bet v 4, and Bet v 6, were analyzed.Results: Among 58 participants, 44 individuals (75.9%) reported PFAS. 32 out of 44 (72.7%) participants reported apple allergy. Bet v 1 exhibited the highest sensitization rate at 82.8%, followed by Bet v 2 (29.3%) and Bet v 6 (1.7%). The combined sensitization rate for Bet v 1 and/or Bet v 2 was 93.1%. A total of 77.6% of the subjects demonstrated sensitization to single component, while 19.0% exhibited sensitization to two components. The sIgE levels of birch pollen and Bet v 1 were significantly elevated in PFAS group compared to non-PFAS group (p=0.001, p< 0.001, respectively), as well as in apple-allergic and non-apple-allergic group (p< 0.001, p< 0.001, respectively). The optimal cut-off values for birch pollen and Bet v 1 sIgE were determined to be 7.09 kUA/L (with a sensitivity of 84.1% and specificity of 78.6%) and 5.11 kUA/L (with a sensitivity of 75.0% and specificity of 85.7%) when diagnosing PFAS. In terms of apple allergy, the optimal cut-off value were 9.40 kUA/L (with a sensitivity of 81.3% and specificity of 76.9%) and 6.53 kUA/L (with a sensitivity of 84.4% and specificity of 84.6%), respectively.Conclusion: The predominant sensitization pattern is mono-sensitization to Bet v 1, but when considering immunotherapy, Bet v 2 should also be taken into account. Bet v 1 serves as a valuable biomarker for diagnosing PFAS and apple allergy.Keywords: birch pollen, allergen component, Bet v 1, pollen food allergy syndrome, apple allergy

Apples are available all year round since they can be stored for long time. However, the expression of the major apple allergen Mal d 1 can increase in the fruit during storage and incease so the risk of allergies in sensitive consumers. Therefore, we studied the change in the expression of the Mal d 1 gene during storage in the modern cultivar UEB 32642 (known under the brand name Opal®) and the cultivar Gala Brookfield (cv. Gala) as a reference. The cultivars were stored under two different conditions, ultra-low oxygen (ULO) and standard cold conditions. The gene expression was evaluated both in fresh fruits and in fruits stored for four, six, and nine months. Nine isoforms of the gene Mal d 1 with the highest published expression were quantified using real-time PCR. The most expressed isoforms Mal d 1.01, Mal d 1.02 and Mal d 1.06A were found in the fresh and also in the stored fruits. The expression of the Mal d 1.03G and Mal d 1.06D isoforms was higher the sored fruits. Our study confirmed that (i) Opal® had a lower overall expression of the Mal d 1 gene than cv. Gala, both in the fresh and stored fruits; (ii) standard cold storage is superior in preserving lower Mal d 1 levels in Opal®apples compared to the ULO conditions; and (iii) less expressed isoforms may be responsible for the general increase in the Mal d 1 gene expression during storage.

Apples are available all year round since they can be stored for long time. However, the expression of the major apple allergen Mal d 1 can increase in the fruit during storage and incease so the risk of allergies in sensitive consumers. Therefore, we studied the change in the expression of the Mal d 1 gene during storage in the modern cultivar UEB 32642 (known under the brand name Opal®) and the cultivar Gala Brookfield (cv. Gala) as a reference. The cultivars were stored under two different conditions, ultra-low oxygen (ULO) and standard cold conditions. The gene expression was evaluated both in fresh fruits and in fruits stored for four, six, and nine months. Nine isoforms of the gene Mal d 1 with the highest published expression were quantified using real-time PCR. The most expressed isoforms Mal d 1.01, Mal d 1.02 and Mal d 1.06A were found in the fresh and also in the stored fruits. The expression of the Mal d 1.03G and Mal d 1.06D isoforms was higher in the stored fruits. Our study confirmed that (i) Opal® had a lower overall expression of the Mal d 1 gene than cv. Gala, both in the fresh and stored fruits; (ii) standard cold storage is superior in preserving lower Mal d 1 levels in Opal®apples compared to the ULO conditions; and (iii) less expressed isoforms may be responsible for the general increase in the Mal d 1 gene expression during storage. [ABSTRACT FROM AUTHOR]

Apples are available all year round since they can be stored for long time. However, the expression of the major apple allergen Mal d 1 can increase in the fruit during storage and incease so the risk of allergies in sensitive consumers. Therefore, we studied the change in the expression of the Mal d 1 gene during storage in the modern cultivar UEB 32642 (known under the brand name Opal®) and the cultivar Gala Brookfield (cv. Gala) as a reference. The cultivars were stored under two different conditions, ultra-low oxygen (ULO) and standard cold conditions. The gene expression was evaluated both in fresh fruits and in fruits stored for four, six, and nine months. Nine isoforms of the gene Mal d 1 with the highest published expression were quantified using real-time PCR. The most expressed isoforms Mal d 1.01, Mal d 1.02 and Mal d 1.06A were found in the fresh and also in the stored fruits. The expression of the Mal d 1.03G and Mal d 1.06D isoforms was higher in the stored fruits. Our study confirmed that (i) Opal® had a lower overall expression of the Mal d 1 gene than cv. Gala, both in the fresh and stored fruits; (ii) standard cold storage is superior in preserving lower Mal d 1 levels in Opal®apples compared to the ULO conditions; and (iii) less expressed isoforms may be responsible for the general increase in the Mal d 1 gene expression during storage. [ABSTRACT FROM AUTHOR]

Interactions between phenolic compounds and the allergen Mal d 1 are discussed to be the reason for better tolerance of apple cultivars, which are rich in polyphenols. Because Mal d 1 is susceptible to proteolytic digestion and allergenic symptoms are usually restricted to the mouth and throat area, the release of native Mal d 1 during the oral phase is of particular interest. Therefore, we studied the release of Mal d 1 under different in vitro oral digestion conditions and revealed that only 6-15% of the total Mal d 1 present in apples is released. To investigate proposed polyphenol-Mal d 1 interactions, various analytical methods, e.g., isothermal titration calorimetry, 1 H- 15 N-HSQC NMR, and untargeted mass spectrometry, were applied. For monomeric polyphenols, only limited noncovalent interactions were observed, whereas oligomeric polyphenols and browning products caused aggregation. While covalent modifications were not detectable in apple samples, a Michael addition of epicatechin at cysteine 107 in r-Mal d 1.01 was observed.

Background: Limited is known on the profiles of apple allergy in China., Objective: To explore the clinical significance of apple allergen components in northern China., Methods: This study recruited 40 participants and categorized into apple tolerance ( n  = 19) and allergy ( n  = 21) group. The latter was categorized into oral allergy symptoms (OAS, n  = 14) and generalized symptoms (GS, n  = 7). All participants underwent ImmunoCAP screening to assess sIgE levels of birch, apple, and their components., Results: The sensitization rates were 90% for Bet v 1, 85% for Mal d 1, 35% for Bet v 2, and 20% for Mal d 3. The overall positive rate for apple allergens was 97.5%, with half demonstrating mono-sensitization to Mal d 1. Birch, Bet v 1 and Mal d 1 sIgE levels had consistent areas under the curve (AUC 0.747, p  = 0.037; AUC 0.799, p  = 0.012; AUC 0.902, p  < 0.001 respectively) in diagnosing apple allergy. The optimal cut-off values were determined to be 22.85 kUA/L (63.6% sensitivity, 85.7% specificity), 6.84 kUA/L (81.8% sensitivity, 71.4% specificity) and 1.61 kUA/L (93.8% sensitivity, 75.0% specificity), respectively. No allergens or components demonstrated diagnostic value in distinguishing between OAS and GS. Mal d 3 sensitization was correlated with mugwort allergy and higher risk of peach, nuts or legumes generalized allergy., Conclusion: Mal d 1 was major allergen and the best for diagnosing apple allergy. Mal d 3 does not necessarily indicate severe allergic reaction to apples in northern China but may indicate mugwort sensitization and an increased risk of peach, nuts or legumes allergy., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Wang, Chen, Lan, Wang and Wang.)

Copyright of Revue Francaise d'Allergologie is the property of Elsevier B.V. and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Copyright of Erwerbs-Obstbau is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Scope: Apples are an important polyphenol (PP) source. To compare the health benefits of traditional and commercial varieties, the phenolic contents and profiles as well as their release from the matrix (bioaccessibility) during oral digestion are determined. Furthermore, based on these data the proposed beneficial effect of PP on the variety specific allergenicity is discussed., Methods and Results: Phenolics are quantified by HPLC-DAD. Total phenolic contents (TPC) are in the range of 111-645 and 343-1950 mg 100 g -1 dry weight for flesh and peel, respectively. Matrix release during oral digestion is investigated ex vivo, with centrifuged and non-centrifuged human saliva and in vitro with simulated saliva fluid (SSF). The overall bioaccessibility is similar in all digestion media, ranging between 40-80% and 39-65% of the TPC in flesh and peel, respectively. Analyzing the correlation among Mal-d 1-allergen-content, unoxidized PP, and the allergenic potential for the samples reveals a negligible effect of phenolics., Conclusion: Due to higher phenolic contents in combination with a similar release, increased PP concentrations in the oral phase and an improved uptake of PP from traditional varieties are assumed. However, the proposed beneficial effect of phenolics on allergenicity cannot be confirmed., (© 2023 The Authors. Molecular Nutrition & Food Research published by Wiley-VCH GmbH.)

About 50–70% of patients allergic to birch pollen suffer from sensitization after apple ingestion. Apple allergenicity was established in only few varieties. Studies were performed on apple fruits of 21 traditional and nine modern varieties organically, intensively, or integratively produced. The aim of the study was to assess whether the factors like cultivation method, maturity stage, genotype, or type of tissue place an impact on the allergenic potential of apples. To answer these questions, we used semiquantitative real-time PCR, ELISA, and immunoblotting. Apple allergen genes present divergent expression across apple cultivars. Expression of the Mal d 1.06A correlates with the Mal d 1 level and is affected by the cultivation method and maturity of the fruit. The content of the main allergen Mal d 1 varied widely across cultivars. Interestingly, in our study, the Gala variety presented a low Mal d 1 concentration regardless of the cultivation method. Based on the Mal d 1.06A expression, the Mal d 1 protein content, and the immunoreactivity assay, the Kandil Sinap, Kosztela, Rumianka from Alma-Ata, Kantówka Gdańska, Reinette Coulon, and Gala cultivars emerged as potentially hypoallergenic apple cultivars. Our study allowed distinguishing between potentially low, medium, and highly allergenic varieties.

Copyright of Acta Alimentaria is the property of Akademiai Kiado and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Mal d 1 is the primary apple allergen in northern Europe. To explain the differences in the allergenicity of apple varieties, it is essential to study its properties and interaction with other phytochemicals, which might modulate the allergenic potential. Therefore, an optimized production route followed by an unsophisticated purification step for Mal d 1 and respective mutants is desired to produce sufficient amounts. We describe a procedure for the transformation of the plasmid in competent E. coli cells, protein expression and rapid one-step purification. r-Mal d 1 with and without a polyhistidine-tag are purified by immobilized metal ion affinity chromatography (IMAC) and fast-protein liquid chromatography (FPLC) using a high-resolution anion-exchange column, respectively. Purity is estimated by SDS-PAGE using an image-processing program (Fiji). For both mutants an appropriate yield of r-Mal d 1 with purity higher than 85% is achieved. The allergen is characterized after tryptic in gel digestion by peptide analyses using HPLC-MS/MS. Secondary structure elements are calculated based on CD-spectroscopy and the negligible impact of the polyhistidine-tag on the folding is confirmed. The formation of dimers is proved by mass spectrometry and reduction by DTT prior to SDS-PAGE. Furthermore, the impact of the freeze and thawing process, freeze drying and storage on dimer formation is investigated.

The allergen Mal d 1 is often responsible for adverse allergic reactions to fresh apples in northern and central Europe. The Mal d 1 content and isoallergen profile are proposed to have an impact on the allergenic potential of the fruit. Therefore, we investigated the impact of the cropping system on the Mal d 1 content and the isoallergen profile of apples by mass spectrometry for the varieties 'Jonagored' and 'Topaz'. To monitor the impact of storage time and conditions, apples of the varieties 'Santana' and 'Jonagold' were stored for up to 12 weeks under regular air (RA), under RA in combination with 1-methylcyclopropene (1-MCP) treatment, and under a controlled atmosphere (CA). The impact of the cropping system (integrated production vs organic production) was negligible. However, a significant increase in the Mal d 1 content during storage was observed, being higher when stored under CA conditions than under RA conditions. An additional treatment with 1-MCP prior to RA storage drastically reduced the level of Mal d 1 expression in the flesh of the apples by ∼50%. Furthermore, the content of isoallergens 1.03 and 1.06 increased disproportionately under CA conditions, while under RA conditions, only isoallergen 1.06 was affected. With the 1-MCP treatment, no changes in the isoallergen profile were obvious.

Fruit and vegetable allergies are prevalent commonly in adults, infants, and children all around the world, but more importantly in Europe and North America. The only solution is complete abstinence from the responsible food, which can be very difficult due to their presence in the form of hidden or undeclared ingredients. Various studies have shown the direct effect of processing on the secondary structure of proteins that can mitigate the allergic properties. The impact of these processing techniques on fruit and vegetable allergies have shown limited success due to the fact that they have multiple allergens that are especially heat stable. Apples, kiwi, peach, and melons are common fruits, whereas celery and carrot are the common vegetables that can result in allergic reactions for some portion of the population upon exposure. Abbreviations: IgE: immunoglobulin E, OAS: oral allergy syndrome, nsLTP: nonspecific lipid transfer proteins, LTP: lipid transfer proteins, HPP: high-pressure processing, PEF: pulsed electric field, CD: circular dichroism, pI: isoelectric point, DBPCFCs: double-blind placebo-controlled food challenges [ABSTRACT FROM AUTHOR]

Allergy to freshly consumed apple fruits is often associated to pollinosis and manifested as oral allergy syndrome (OAS). The allergenic properties of apple varieties differ greatly, spanning from low allergenic to high allergenic varieties. The knowledge of the genetic determinants for allergenicity has been of great interest in scientific community for several years, but the molecular mechanisms involved are still little understood. Here, factors putatively involved in allergenicity were investigated at biochemical and molecular level in pollen and in fruits of apple varieties differing in their allergenic potential. Among putative sensitizing factors, transglutaminase (TGase) and phospholipase A 2 (PLA 2 ) were considered together with reactive oxygen species (ROS) and known apple allergen genes, with particular attention devoted to the Mal d 1 gene family, the most important one in sensitization. We found that the expression of some allergen genes and the activities of TGase, PLA 2 and ROS producing enzyme are lower in the hypo-allergenic variety ‘Durello di Forlì’ in comparison with the high-allergenic genotypes ‘Gala’ and ‘Florina’. These results highlight correlations among allergen expressions, enzymatic activities and apple cultivars; these data underline the possibility that some of them could be used in the future as markers for allergenicity. [ABSTRACT FROM AUTHOR]

Background Cross-reactive apple allergy is a common co-morbidity of birch pollen allergy, caused by the presence of a Bet v 1 homologue allergen in apple, Mal d 1. Treatment of tree pollen hay fever by immunotherapy is well established, but its effect on the accompanying apple allergy is debated. Objective To establish a mouse model of birch pollen induced cross-reactivity to Mal d 1 and investigate the effect of birch pollen immunotherapy on the cross-reactivity to Mal d 1. Methods Respiratory allergy was induced in Balb/c mice by intraperitoneal exposure to alum-adsorbed birch pollen extract ( BPE) in combination with short or prolonged intranasal exposure to BPE. To evaluate the response to Mal d 1, mice were exposed intraperitoneally to Mal d 1. Immunoglobulin responses and cytokine production by splenocytes were measured by ELISA. Allergic symptoms were evaluated by measuring airway hyper-reactivity and hypothermia as a surrogate marker for anaphylaxis. Immunotherapy was performed subcutaneously with alum-adsorbed BPE. Results Mice exposed to BPE develop cross-reactive IgE to Mal d 1. Early after exposure to BPE, this response is still weak and does not yet translate into anaphylaxis. Interestingly, later re-challenge with BPE increased cross-reactivity to a level where Mal d 1 exposure induced anaphylaxis. Cross-sensitization can also be induced by systemic Mal d 1 exposure. Birch pollen immunotherapy significantly reduced the anaphylactic response of mice to Mal d 1. Conclusion & Clinical Relevance A mouse model mimicking birch pollen induced cross-reactivity to Mal d 1 was successfully established. In this model, birch pollen immunotherapy significantly ameliorated the anaphylaxis induced by Mal d 1. Our experimental data suggest that boosting of Mal d 1 recognizing immunoglobulins by BP SCIT is important for the amelioration of apple allergy in human. [ABSTRACT FROM AUTHOR]

The apple allergy in Northern Europe is a cross-reaction to the birch pollen allergy. No correlation between the allergenicity of an apple variety and the content of the major apple allergen Mal d 1, a homologue to the Bet v 1 allergen in birch, could be found using ELISA, so far. Therefore, an impact of polyphenols and/or differences in the isoallergen profile are discussed. To allow a more detailed analysis of the Mal d 1 content and the isoallergen profile, a mass spectrometric method was applied to investigate differences in the flesh and peel of 10 traditional varieties and 10 commercial breeds. The data revealed often, but not always, lower Mal d 1 contents in traditional varieties grown in orchard meadows, which was more obvious in the flesh. Differences among the peels were less pronounced. A closer look at the individual isoallergens 1.01, 1.02, 1.03, and 1.06 reveals an increased impact of the minor isoallergens 1.03 and 1.06 on the allergenic potential, since commercial breeds like Braeburn, Santana, and Holstein Cox, which are considered to have reduced allergenic potentials, were characterized by low levels of these isoallergens.

Background Genetic modification of allergenic foods such as apple has the potential to reduce their clinical allergenicity, but this has never been studied by oral challenges in allergic individuals. Methods We performed oral food challenges in 21 apple-allergic individuals with Elstar apples which had undergone gene silencing of the major allergen of apple, Mal d 1 , by RNA interference. Downregulation of Mal d 1 gene expression in the apples was verified by qRT- PCR. Clinical responses to the genetically modified apples were compared to those seen with the wild-type Elstar using a visual analogue scale ( VAS). Results Gene silencing produced two genetically modified apple lines expressing Mal d 1.02 and other Mal d 1 gene mRNA levels which were extensively downregulated, that is only 0.1-16.4% (e- DR1) and 0.2-9.9% (e- DR2) of those of the wild-type Elstar, respectively. Challenges with these downregulated apple lines produced significantly less intense maximal symptoms to the first dose (Vmax1) than with Elstar (Vmax1 Elstar 3.0 mm vs 0.0 mm for e- DR1, P = 0.017 and 0.0 mm for e- DR2, P = 0.043), as well as significantly less intense mean symptoms per dose (meanV/d) than with Elstar (meanV/d Elstar 2.2 mm vs 0.2 mm for e- DR1, P = 0.017 and 0.0 mm for e- DR2, P = 0.043). Only one subject (5%) remained symptom-free when challenged with the Elstar apple, whereas 43% did so with e- DR1 and 63% with e- DR2. Conclusion These data show that mRNA silencing of Mal d 1 results in a marked reduction of Mal d 1 gene expression in the fruit and reduction of symptoms when these apples are ingested by allergic subjects. Approximately half of the subjects developed no symptoms whatsoever, and virtually all subjects wished to consume the apple again in the future. [ABSTRACT FROM AUTHOR]

BACKGROUND Up to 6.6% of adults and 11.5% of children aged between 0 and 6 years are reporting a perceived fruit allergy. In the last group 8.5% perceived reactions against apple. To keep allergic reactions as low as possible and to make apple consumption more tolerable for patients allergic to apple, the Mal d 1 content of four apple cultivars ('Boskoop', 'Elise', 'Pinova', 'Santana' and 'Topaz') were analyzed, to identify apple cultivars with only a low Mal d 1 content. Further, the development of Mal d 1 content of low-allergenic cultivars should be investigated during long-term storage. RESULTS The lowest Mal d 1 content was measured for fruits of the cultivar 'Santana' at harvest, followed by 'Pinova' and 'Elise' after storage without 1- MCP. Nearly all fruits treated with 1- MCP had a significantly lower Mal d 1 content than fruits without 1- MCP, except for 'Boskoop' fruits after 44 weeks of storage. Apple cultivars with a low Mal d 1 content are not a cross of 'Golden Delicious' with another cultivar, with the exception of 'Pinova' which is a cultivar from the so-called 'Pi-series', which have a moderate resistance to scab and mildew. First experiments on tomatoes and peanuts have shown a reduction of allergic proteins in older and child generations after transfer of RNA. This reduction is probably based on an alteration of gene expression regulating the synthesis of allergic proteins, leading to changes in protein composition in the fruit. CONCLUSION Individuals allergic to apple should choose the described cultivars with relative low Mal d 1 content. If breeders want to breed cultivars with low allergy, they should avoid crossings with 'Golden Delicious'. © 2013 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Background Birch pollen allergies are frequently associated with adverse reactions to various fruits, nuts, or vegetables, described as pollen-food syndrome ( PFS) and caused by cross-reactive IgE antibodies primarily directed against Bet v 1. Specific immunotherapy ( SIT) represents an effective treatment for inhalant allergies; however, successful birch pollen SIT does not correlate well with the amelioration of concomitant food allergies. Methods As vaccine candidates, apple Mal d 1 as well as hazelnut Cor a 1 derivatives were designed by in silico backbone analyses of the respective allergens. The proteins were produced by site-directed mutagenesis as fold variants of their parental allergens. Because Mal d 1 and Cor a 1 form cysteine-mediated aggregates, nonaggregative cysteine to serine mutants were also generated. The proteins were characterized physicochemically, immunologically, and in in vivo models with or without adjuvant. Results The structurally modified proteins showed significantly decreased IgE binding capacity. Notably, both in vivo models revealed reduced immunogenicity of the hypoallergenic fold variants. When formulated with alum, the monomeric cysteine mutants induced a similar immune response as the aggregated parental allergens, which is in contrast with data published on Bet v 1. Conclusion These findings lead to the suggestion that the Bet v 1 structure has unique intrinsic properties, which could account for its high allergenicity. Obviously, these characteristics are not entirely shared with its food homologues from apple and hazelnut. Thus, it is important to tackle pollen-related food allergies from different angles for the generation of effective vaccine candidates to treat birch PFS. [ABSTRACT FROM AUTHOR]

Patients with Oral Allergy Syndrome (OAS) to fresh apple may tolerate low allergenic apple cultivars. We aimed to investigate if the low allergenic properties of Elise and Santana, as previously identified in a Dutch population, could be generalised within North West Europe within the birch pollen region with regard to both the prevalence and degree of sensitization. Prick-to-prick tests (PTP) were performed in eighty-five adult patients with OAS to fresh apple in Great Britain, Switzerland and Northern Italy, before the birch pollen season, using the putatively low allergenic apple cultivars Elise, Santana, Granny Smith, Modi and Mcintosh, as well as the putatively high allergenic apple cultivars Golden Delicious and Kanzi. No significant differences in percentages of negative responses of PTPs were found between the three countries. Negative responses did not differ from negative responses to the different apple cultivars we previously found in 2006/2007 in the Netherlands. The size of the PTPs of all apple cultivars tested were correlated to the size of the skin prick tests with birch pollen. These results add to the indications for the low allergenic properties of the low allergenic apple cultivars Santana and Elise, as the number of negative responses were reproducible in three countries within the birch pollen region and were similar to previous results in the Netherlands. These results justify oral challenge studies with Elise and Santana within the birch pollen region, to establish the low allergenic properties for the benefit for apple allergic consumers for definite conclusions. [ABSTRACT FROM AUTHOR]

Patients who suffer from birch pollinosis often develop adverse reactions to the consumption of fresh apples due to the structural similarity of the allergens Bet v 1 and Mal d 1 from birch and apples, respectively. A different allergenic potential for Mal d 1 isoallergens is postulated, but approaches to quantify the Mal d 1 isoallergen-specific are missing. Therefore, a bottom-up proteomics approach was developed to quantify Mal d 1 by stable isotope dilution and microHPLC-QTOF analyses. Marker peptides for individual isoallergens (Mal d 1.01-Mal d 1.03 and Mal d 1.06), combinations thereof (Mal d 1.01 + 1.02, Mal d 1.02 + 1.06, and Mal d 1.04 + 1.05), and two global marker peptides, comprising Mal d 1.01 + 1.02 + 1.04 + 1.05 and Mal d 1.03 + 1.06 + 1.07 + 1.08 + 1.09, were identified. By the use of an extraction standard (r-Mal d 1&#95;mut), an optimized protocol for extraction and tryptic digestion of apple proteins was developed, and the variety-specific extraction efficiency was monitored for the flesh and peel of apples. The Mal d 1 contents in flesh and peel of five commercial apple breeds and four apple varieties from orchard meadows were quantified isoallergen-specific.

The consumption of apple fruits can induce allergic reactions. The major allergen responsible for these allergic reactions in Central Europe and North America is the apple allergen Mal d 1, the synthesis of which depends on the apple cultivar and increases during storage time. Several studies have investigated the impact of the cultivar and storage conditions on the Mal d 1 content in apple fruits. However, there are few data available on the impact of 1-methylcyclopropene (1-MCP) on Mal d 1 synthesis during long-term storage. In this study, the Mal d 1 content of seven different apple cultivars was analyzed. Half of the fruits were treated with 1-MCP (Smart Fresh™, Agro Fresh Inc. Spring House, PA, USA). After treatment, the fruits were stored for 12, 20, 28 and 36 weeks in a cold chamber at 2 °C as well as for 12, 20, 28, 36 and 44 weeks under controlled atmosphere (CA-storage, 1.5% CO2, 1.5% O2, 2 °C). The Mal d 1 contents of all apple cultivars increased during storage. With the exception of ‘Boskoop’, Mal d 1 contents of all of the fruits treated with 1-MCP were significantly lower than those of the fruits in the untreated control group. [ABSTRACT FROM AUTHOR]

Abstract: The most important apple allergen in Central Europe and North America is Mal d 1. Apples are a very important source of secondary plant metabolites like polyphenols in human nutrition. It is known that oxidised phenols can bind proteins. These irreversible bindings can lead to a reduced allergenicity. The most important phenols in apple are epicatechin, catechin and their polymeric structures, which have been identified as substrates of the polyphenoloxidase (PPO). The aim of this study was to analyse the influence of naturally occurring catechin and epicatechin contents in apple on the allergenicity of apple fruits. Fruits of the cultivars ‘Elstar’, ‘Diwa’ and ‘Boskoop’ were harvested and stored for 8 and 12weeks in a cold-chamber at 2°C. Mal d 1-, catechin-, epicatechin- and total phenol content as well as the activity of PPO were determined. Correlation analysis showed that naturally occurring catechin as well as epicatechin has no impact on the Mal d 1 content of the tested cultivars: correlation coefficient ranged from −0.203 to 0.501 for the correlation between Mal d 1 and catechin. The results further indicated that the activity of PPO is more important than the content of total phenols to reduce the Mal d 1 level. If there is a high PPO activity, Mal d 1 could be reduced even if the total phenol concentration is low. [Copyright &y& Elsevier]

Background: A considerable number of individuals suffer from oral allergy syndrome (OAS) to apple, resulting in the avoidance of apple consumption. Apple cultivars differ greatly in their allergenic properties, but knowledge of the causes for such differences is incomplete. Mal d 1 is considered the major apple allergen. For Mal d 1, a wide range of isoallergens and variants exist, and they are encoded by a large gene family. To identify the specific proteins/genes that are potentially involved in the allergy, we developed a PCR assay to monitor the expression of each individual Mal d 1 gene. Gene-specific primer pairs were designed for the exploitation of sequence differences among Mal d 1 genes. The specificity of these primers was validated using both in silico and in vitro techniques. Subsequently, this assay was applied to the peel and flesh of fruits from the two cultivars 'Florina' and 'Gala'. Results: We successfully developed gene-specific primer pairs for each of the 31 Mal d 1 genes and incorporated them into a qRT-PCR assay. The results from the application of the assay showed that 11 genes were not expressed in fruit. In addition, differential expression was observed among the Mal d 1 genes that were expressed in the fruit. Moreover, the expression levels were tissue and cultivar dependent. Conclusion: The assay developed in this study facilitated the first characterisation of the expression levels of all known Mal d 1 genes in a gene-specific manner. Using this assay on different fruit tissues and cultivars, we obtained knowledge concerning gene relevance in allergenicity. This study provides new perspectives for research on both plant breeding and immunotherapy. [ABSTRACT FROM AUTHOR]

Copyright of Erwerbs-Obstbau is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

To evaluate differences in skin prick test response to apple cultivars in patients with apple allergy, 19 patients (10 adults and 9 children) underwent prick-to-prick skin prick test with eleven commercial and noncommercial apple cultivars, and evaluation of specific IgE to apple and recombinant apple allergens Mal d 1 and Mal d 3. The results show that different reactions might be evoked in a single patient by different apple cultivars and also separately for the peel and the pulp of a single cultivar. The cultivars were ranked according to their allergenicity level, being Jonathan, Jonagold, Golden Delicious and Fiesta the most allergenic when considering the pulp and Gala, Fiesta and Golden Delicious for the peel. Further investigations are needed to clarify if a single patient can be allergic only to well-defined apple cultivars and which allergometric tests are necessary to ascertain this. [ABSTRACT FROM AUTHOR]

Abstract: Lipid transfer protein (LTP) has been reported as an important allergen inducing severe systemic reactions in allergic subjects. The aim of the present study was to estimate the role of LTP in diagnosis of apple allergy in children allergic to birch pollen. The prospective study was carried out on a group of 21 children sensitized to birch pollen and apple allergens, confirmed by presence of specific IgE in the sera of patients. Allergens separated by electrophoresis were probed by Western blotting using hypersensitive patients’ sera. Food hypersensitivity to apple was verified by oral food challenge test. Six patients’ sera were hypersensitive to birch pollen and apple proteins. Almost all sera recognized specifically the main allergen of apple peal Mal d 3 with molecular weight <10kDa (LTP). Positive oral challenge to apple was found in 52.4% of investigated children. Children allergic to Mal d 1 presented different clinical symptoms. Sensitization to Mal d 3 is very common in children with apple allergy. No correlation between clinical symptoms and sensitization to recombinant apple allergens has been found, LTP may be useful in diagnosis of apple allergy. [Copyright &y& Elsevier]

Background: Individual amino acid residues of the major birch pollen allergen, Bet v 1, have been identified to be crucial for IgE recognition. The objective of the present study was to evaluate whether this concept was applicable for the Bet v 1-homologous apple allergen, Mal d 1. Methods:A Mal d 1 five-point mutant was produced by PCR techniques, cloned into pMW 172 and expressed in Escherichia coli BL21(DE3) cells. To evaluate the allergenic properties of the engineered protein compared to Mal d 1 wild-type IgE immunoblotting, ELISA, peripheral blood monocytes proliferation assays, and skin prick tests were performed. Results: The Mal d 1 mutant showed reduced capacity to bind specific IgE as compared to wild-ype Mal d 1 in in vitro assays in the majority of the sera tested. In ELISA, 10 out of 14 serum samples displayed an 88-30% decrease in gE binding to Mal d 1 mutant compared to wild-type Mal d 1. Skin prick tests in apple-allergic patients (n = 2) confirmed the markedly decreased ability of the Mal d 1 mutant to induce allergic reactions in vivo. However, the relevant T cell epitopes were present in the mutated molecule according to peripheral blood mononuclear cell proliferation assays. Conclusions: Our findings suggest that it is possible to modulate the IgE-binding properties of allergens by single amino acid substitutions at crucial positions which might be useful for future immunotherapy of birch-pollen-associated food allergies which are not ameliorated by birch pollen immunotherapy. [ABSTRACT FROM AUTHOR]

Background: Lipid transfer proteins (LTP) are stable, potentially life-threatening allergens in fruits and many other vegetable foods. The aim of this study was to clone and express recombinant apple LTP (Mal d 3), as has previously been done for peach LTP (Pru p 3) and set up quantitative tests for measuring fruit LTPs. Methods: cDNA for Mal d 3 and Pru p 3 was cloned, expressed in the yeast Pichia pastoris and the resulting proteins were purified via cation exchange chromatography. The immune reactivity of rMal d 3 was compared to nMal d 3 by RAST (inhibition), immunoblotting and basophil histamine release testing. To obtain monoclonal and monospecific polyclonal antibodies, mice and rabbits were immunized with purified nMal d 3. Results: The deduced amino acid sequence of Mal d 3 was identical to the published sequence, Pru p 3 differed at two positions (S9A and S76H). The rMal d 3 had an IgE-binding potency and biological activity close to its natural counterpart. One sandwich ELISA selectively detecting apple LTP and another cross-reactive with cherry, nectarine and hazelnut LTP were developed. In addition, a competitive RIA was developed with polyclonal rabbit antiserum and labeled nMal d 3. Conclusion: rMal d 3 (as shown before for rPru p 3) may be a useful tool for application in component-resolved diagnosis of food allergy. Assays for the measurement of LTP will increase the traceability of this potentially dangerous allergen. Copyright © 2005 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

The true prevalence of food allergy as an IgE mediated reaction is still under discussion. Using apple as a model allergen source a multidisciplinary consortium worked together at developing various strategies for reducing the incidence of fruit allergies in an EC-funded project. Patient allergen profiles were established using in vitro and in vivo tests with respect to geographic area and mild or severe symptoms. Apple allergens (Mal d 1–Mal d 4) were characterised, variants identified, cloned and sequenced. These individual allergens were used to increase the sensitivity and specificity of diagnosis. Furthermore, they provided better prognosis of disease severity. RT-PCR and ELISA were developed for determining the allergen specific mRNA and expressed allergenic protein in a large number of apple cultivars. Similarly, changes in allergen characteristics from harvest through storage to processing and the impact of agronomic practices were investigated. Allergen genes were mapped on a molecular linkage map of apple. The biological function of Mal d 1 was studied using the RNA interference strategy. Finally, consumer attitudes in Northern, Central and Southern Europe were gauged on the acceptability of low allergen cultivars or a GMO and its impact on product quality. [ABSTRACT FROM AUTHOR]

Background: Recent studies showed that injection specific immunotherapy (SIT) with birch pollen extract greatly reduces or cures the associated apple allergy in a large proportion of birch pollen-allergic patients. However, the long-term efficacy of SIT for apple allergy has not been assessed. Objective: To evaluate the duration of the effect of injection SIT with birch pollen extract on apple allergy in birch pollen-allergic patients. Methods: Thirty birch pollen-allergic patients showing both the clinical disappearance of apple allergy and a negative SPT with fresh apple at the end of their injection SIT course were followed-up at 12-month intervals from 6 months after SIT was stopped. Apple tolerance as well as SPT was assessed on all occasions. Fifty-seven birch pollen-allergic subjects without apple allergy and not submitted to SIT regularly followed-up for the onset of oral allergy syndrome (OAS) were used as controls. Results: The overall prevalence of OAS after 30 months of follow-up did not differ between patients and controls. Although most patients became re-sensitized to apple by SPT over time, >50% of them were still able to tolerate eating the fruit at the 30-month follow-up visit. Conclusion: Although most patients show a ‘natural’, gradual propensity to apple re-sensitization (a consequence of prolonged and repeated inhalation of birch pollen responsible for primary sensitization?), the clinical effects of injection SIT on food allergy seem rather long lasting. [ABSTRACT FROM AUTHOR]

BackgroundMost patients with birch pollen allergy report oral allergy symptoms after eating fresh apples and other vegetable foods. Major birch pollen and apple allergens, Bet v 1 and Mal d 1, are highly homologous; as a consequence, pollen-specific immunotherapy (SIT) might be expected to improve apple hypersensitivity. ObjectiveTo evaluate the clinical and immunological effects of birch pollen SIT on oral allergy syndrome (OAS) induced by apples. MethodsA prospective study carried out in 49 birch pollen-sensitive patients with apple-induced OAS who received injection immunotherapy for 12, 24, or 36 months. Twenty-six patients not submitted to SIT and followed up for 12–48 months were used as controls. Both SPT and open oral challenges with fresh golden delicious apple were performed, as well as specific IgE measurements, before and after SIT. ResultsForty-one patients (84%) vs no control (0%) reported a significant reduction (50–95%) or a total disappearance (100%) of OAS symptoms after SIT (P < 0.001). Similar responses were observed in patients treated for 12, 24, or 36 months. SIT also induced a marked reduction in skin reactivity against fresh apple in 43 patients (88%). The effect of SIT was inversely related with baseline skin reactivity: 50% and 8% patients with a weakly or strongly positive baseline apple skin prick tests (SPT), respectively, did not report changes in OAS severity after SIT (P < 0.01). In contrast, baseline birch pollen-specific or apple-specific IgE antibodies levels did not influence SIT effectiveness on OAS. SIT induced a marked decrease in birch pollen-specific IgE levels (P < 0.001), whereas apple-specific IgE showed an unexpected variability (reduction in 21%, no change in 43%, increase in 38%). No control subject reported a reduction in OAS severity or showed a decrease in skin reactivity at follow-up (P < 0.001). ConclusionsSIT with birch pollen extracts effectively reduces clinical apple sensitivity and skin... [ABSTRACT FROM AUTHOR]

The major allergen from birch pollen, Bet v I, and the cross-reacting 18-kDa major allergen from Golden Delicious and Granny Smith applies were isolated by micropreparative SDS-PAGE followed by electroelution. In the case of apples, highly active, low-temperature extracts were used. The purity of the allergens was checked by analytic SDS-PAGE and immunoblotting with allergic patients' sera, as well as by N-terminal amino acid microsequencing, and the allergens were found to be very pure. The strong immunologic activity of the isolates was determined by the enzyme allergosorbent test (EAST) and EAST inhibition assays; this activity was, in the case of Bet v I, similar to that of a preparation obtained by monoclonal antibody affinity chromatography. The allergenic potency of Bet v I and of the cross-reactive apple allergen was determined by EAST inhibition and dose-related histamine release. With both assay systems, the allergenic reactivity of Bet v I was considerably higher than that of the major apple allergen. Furthermore, skin prick tests with the purified allergens and with whole allergenic extracts were performed on a group of 33 patients suffering from birch-pollen and apple hypersensitivity, and on a control group of 10 patients. The frequency of positive prick test results in the allergic patient group ranged from 73% for the major allergen from Golden Delicious apples to 97% with Bet v I and whole birch pollen extract, respectively. In contrast to our low-temperature extracts, commercial prick test solutions of four different manufacturers were found to be unreliable for the diagnosis of apple allergy. The skin test results again indicated the strong immunologic activity of the allergen isolates and the predominance of the major allergens in context with birch-pollen and apple hypersensitivity. Taken together, the results support the view that the 18-kDa major allergen represents most of the allergenicity of the the apple fruit, and that all allergenic epitopes of the apple proteins are present on Bet v I. [ABSTRACT FROM AUTHOR]

Skin sensitivity to apple, carrot and potato, clinically related to birch pollinosis was investigated. Different skin test techniques using fresh fruit were compared. A simple prick test (SPT) technique with a lancet piercing the apple peel just before pricking the skin was shown to be the most practical and to give reproducible results. The allergenic activity in apple was found to be heat labile and deteriorated during storage at room temperature. Apple, carrot, potato, hazelnut and birch reactivity was transferable in Prausnitz-Küstner test like IgE antibodies. SPT reactivity to fresh material from apple, carrot and potato was investigated in 174 children of whom 128 suffered from pollen allergy. Positive SPT results were obtained almost exclusively in children who were SPT positive to a birch pollen extract. Children who noticed clinical symptoms when eating apple, raw carrot or potato were found to have a significantly large SPT reaction than children with a negative history. [ABSTRACT FROM AUTHOR]

About 50–70% of patients allergic to birch pollen suffer from sensitization after apple ingestion. Apple allergenicity was established in only few varieties. Studies were performed on apple fruits of 21 traditional and nine modern varieties organically, intensively, or integratively produced. The aim of the study was to assess whether the factors like cultivation method, maturity stage, genotype, or type of tissue place an impact on the allergenic potential of apples. To answer these questions, we used semiquantitative real-time PCR, ELISA, and immunoblotting. Apple allergen genes present divergent expression across apple cultivars. Expression of the Mal d 1.06A correlates with the Mal d 1 level and is affected by the cultivation method and maturity of the fruit. The content of the main allergen Mal d 1 varied widely across cultivars. Interestingly, in our study, the Gala variety presented a low Mal d 1 concentration regardless of the cultivation method. Based on the Mal d 1.06A expression, the Mal d 1 protein content, and the immunoreactivity assay, the Kandil Sinap, Kosztela, Rumianka from Alma-Ata, Kantówka Gdańska, Reinette Coulon, and Gala cultivars emerged as potentially hypoallergenic apple cultivars. Our study allowed distinguishing between potentially low, medium, and highly allergenic varieties. [ABSTRACT FROM AUTHOR]

Mal d 1 is not equally distributed over the apple. We aimed to examine the influence of the location of pricking in the apple on prick-to-prick skin prick test ( PTP) results. PTPs were performed in autumn 2007 and spring 2008, before the birch pollen season, in 32 Dutch adults with symptoms of oral allergy to fresh apple, using apples harvested in autumn 2007. PTPs with fresh intact and unpeeled Pink Lady, Golden Delicious, Elise, Santana and Modi apples were performed using material obtained from approximately 2 cm near the stalk (top), and the middle region. All PTP responses were greater when performed with apple material near the stalk than from the middle region. In 2007, these differences were statistically significant for Pink Lady, Golden Delicious and Elise, and in 2008, for Pink Lady and Modi. When performing PTPs, the apple should be pricked near the stalk rather than in the middle. [ABSTRACT FROM AUTHOR]

• Apple allergy is a well-characterized fruit allergies linked to pollinosis. • Concerning type and amount of allergens apple cultivars show broad biodiversity. • Five Mal d 1 isoallergens were tested for the first time against allergic patient's sera. • Besides apple varieties, differences among isoallergens and patients were found. Apple-related allergy is increasing among European population. Avoidance of apple fruit consumption is the only strategy for sensitized people, with a subsequent shortage of healthy nutrients in the diet. Four different protein families, Mal d 1 to 4, determine the allergy to apple causing syndromes known as oral allergy syndrome (OAS) or gastro-intestinal/generalized syndrome (GS) in sensitized patients. With the aim to highlight the dissimilarities in the immunological response inside the Mal d 1 family, recognized as main determinant for OAS, single purified allergens were tested against sera of paediatric and adult patients suffering for both syndromes. Here, we found that Mal d 1 isoallergens showed a remarkable different allergenicity potential, thus indicating an individual pattern in each patient and a diverse sensitization between age groups. Moreover, two different allergens, Mald 2.01 and Mal d 4.01, were analyzed with the above sera. In addition to patients and allergens variability, differences in amount and localization of the allergens, detected by patient's sera, were revealed among hypo- and hyper allergenic apple varieties, confirming the complexity of this food allergy. [ABSTRACT FROM AUTHOR]

Mal d 1 is the primary apple allergen in northern Europe. To explain the differences in the allergenicity of apple varieties, it is essential to study its properties and interaction with other phytochemicals, which might modulate the allergenic potential. Therefore, an optimized production route followed by an unsophisticated purification step for Mal d 1 and respective mutants is desired to produce sufficient amounts. We describe a procedure for the transformation of the plasmid in competent E. coli cells, protein expression and rapid one-step purification. r-Mal d 1 with and without a polyhistidine-tag are purified by immobilized metal ion affinity chromatography (IMAC) and fast-protein liquid chromatography (FPLC) using a high-resolution anion-exchange column, respectively. Purity is estimated by SDS-PAGE using an image-processing program (Fiji). For both mutants an appropriate yield of r-Mal d 1 with purity higher than 85% is achieved. The allergen is characterized after tryptic in gel digestion by peptide analyses using HPLC-MS/MS. Secondary structure elements are calculated based on CD-spectroscopy and the negligible impact of the polyhistidine-tag on the folding is confirmed. The formation of dimers is proved by mass spectrometry and reduction by DTT prior to SDS-PAGE. Furthermore, the impact of the freeze and thawing process, freeze drying and storage on dimer formation is investigated.