Cabbage lipid transfer protein Bra o 3 is a major allergen responsible for cross-reactivity between plant foods and pollens

doi:10.1016/j.jaci.2006.01.026

Journal of Allergy and Clinical Immunology

Volume 117, Issue 6, June 2006, Pages 1423-1429

https://doi.org/10.1016/j.jaci.2006.01.026 Get rights and content

Background

Food IgE-mediated allergy to members of the Brassicaceae family has been increasingly reported.

Objective

To characterize cabbage—Brassica oleracea var capitata—allergy and its major allergens.

Methods

A prospective study was performed, recruiting 17 patients allergic to cabbage, and control subjects. Skin prick tests and double-blind placebo-controlled food challenges were performed. A major allergen was isolated from cabbage by RP-HPLC and characterized by N-terminal amino acid sequencing and matrix-assisted laser desorption/ionization mass spectrometry analysis. Specific IgE determinations, IgE immunoblots, and CAP-inhibition assays were also performed.

Results

Skin prick test and specific IgE were positive to cabbage in all patients. Five of them referred anaphylactic reactions when eating cabbage, and in another 5 patients, cabbage allergy was further confirmed by double-blind placebo-controlled food challenge. Most of them showed associated sensitizations to mugwort pollen, mustard, and peach. A 9-kd cabbage IgE-binding protein, Bra o 3, was identified as a lipid transfer protein (LTP) with 50% of identity to peach LTP Pru p 3. Skin prick test with Bra o 3 showed positive results in 12 of 14 cases (86%). On CAP inhibition assays, Bra o 3 managed to inhibit significantly the IgE binding to cabbage, mugwort pollen, and peach. Both Bra o 3 and Pru p 3 were recognized by IgE from the patients' sera.

Conclusion

Bra o 3, a cabbage LTP, is a major allergen in this food, cross-reacting with mugwort pollen and with other plant foods, such as peach.

Clinical implications

Cabbage IgE-mediated allergy is a potentially severe condition that can present with other plant food and pollen allergies.

Section snippets

Selection of patients and control subjects

A prospective study was designed in our Allergy Section, composed of a detailed clinical history, in vivo tests, and blood drawing for in vitro assays. During a 2-year running period, every patient reporting immediate adverse reactions related to cabbage ingestion, suggestive of being IgE-mediated, and showing positive skin prick test (SPT) to fresh cabbage, was included. A control group of 17 subjects allergic to dust mite, nonsensitized to either foods or pollens, matched for age and sex with

General characteristics

Seventeen cabbage-hypersensitive patients were included in the study, whose general characteristics are summarized in Table I. Mean age (±SD) was 30.7 ± 5.9 years, ranging from 17 to 43, and with a predominance of female subjects (11:6). Sixteen patients (94.1%) were atopic, as defined by a personal history of respiratory allergy, together with positive SPT to several common aeroallergens, including mugwort pollen in all but 1 case. Out of the atopic group, 7 patients (41.2%) had rhinitis and

Discussion

Regarding food IgE-mediated allergy, it is now well known that cross-reactions among allergens from different sources, as a result of similarities among their IgE epitopes, are responsible for very significant clinical disorders. A clear example is the pollen-food allergy syndrome, which is the most common food hypersensitivity in the adult population, and includes several distinct clinical associations, such as birch pollen with Rosaceae fruit, ragweed pollen with melon, and mugwort pollen

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Research on reported food-related triggers of atopic disease in South Asian adults is lacking despite the region’s large population and the global significance of allergic diseases.
The study aimed to identify prevalent local food items and assess allergic sensitization rates to potential trigger foods for atopic diseases via skin prick and specific IgE testing.
The study began with a pilot survey of 100 subjects recruited from 4 hospitals in Hyderabad, India, focusing on foods perceived to relate to asthma, allergic rhinitis, atopic dermatitis, urticaria, and gastrointestinal allergic symptoms. A subsequent main study evaluated 2010 participants, 1754 of whom were diagnosed with an aforementioned atopic disease and who reported allergic symptoms related to any of 77 foods identified in the pilot study. Ultimately 1622 patients who consented to skin prick and specific IgE testing and who reported at least 1 food item triggering allergic diseases were included in the final analysis.
Among 1622 patients (average age, 42.6 ± 12.9 years; 55.5% male), asthma was the most commonly diagnosed atopic disease (26.4%), with itching and rash being frequently reported symptoms (22.7%). Notably, 94.9% of patients had total serum IgE > 144 kU/L. Chickpea, cabbage, eggplant, walnut, cumin, and betel leaf were the most commonly reported trigger foods.
In this sample of South Indian adults diagnosed with allergic disease, reported food triggers were most commonly local dietary staples, while reported reactions to priority allergens like peanut and sesame were conspicuously absent. Observed concordance between patient-reported food triggers and sensitization to reported food triggers was low, highlighting the need for improved clinical evaluation of suspected triggers.
Plant non-specific lipid transfer proteins: An overview
2022, Plant Physiology and Biochemistry
Citation Excerpt :
It is also interesting to mention that reactive nsLTPs have been also detected in certain pollen (Skypala et al., 2019; Li et al., 2020), which suggests an additional role as aeroallergens. Those of mugwort (Artemisia vulgaris) and plane tree (Platanus acerifolia) show more than 40% sequence identity with Prunus persica's major allergen, Pru p 3, and a cross-reaction with homologous members of certain foods, being therefore potentially responsible for cross-sensitization (Palacín et al., 2006; Uasuf et al., 2020). Besides, corn nsLTP (Zea m 14) is a major allergen responsible for food allergy reactions (Brenna et al., 2000) and barley nsLTP is a relevant allergen in beer (García et al., 2004).
Plant non-specific lipid transfer proteins (nsLTPs) are usually defined as small, basic proteins, with a wide distribution in all orders of higher plants. Structurally, nsLTPs contain a conserved motif of eight cysteines, linked by four disulphide bonds, and a hydrophobic cavity in which the ligand is housed. This structure confers stability and enhances the ability to bind and transport a variety of hydrophobic molecules. Their highly conserved structural resemblance but low sequence identity reflects the wide variety of ligands they can carry, as well as the broad biological functions to which they are linked to, such as membrane stabilization, cell wall organization and signal transduction. In addition, they have also been described as essential in resistance to biotic and abiotic stresses, plant growth and development, seed development, and germination. Hence, there is growing interest in this family of proteins for their critical roles in plant development and for the many unresolved questions that need to be clarified, regarding their subcellular localization, transfer capacity, expression profile, biological function, and evolution.
Molecular aspects of pollen allergens: Where do we currently stand?
2019, Revue Francaise d'Allergologie
Les allergènes polliniques appartiennent à différentes familles de protéines PR (PR pour Pathogenesis-Related), protéines impliquées dans les mécanismes de défense développés par les plantes : PR-2 ou endo β-glucanases, PR-3 ou chitinases, PR-5 ou thaumatin-like protéines (TLP), PR-10 ou protéines Bet v 1-like, PR-12 ou défensines et PR-14 ou protéines de transfert des lipides (LTP). D’autres allergènes polliniques correspondent à des protéines à rôle structural (profilines, expansines), métabolique (polcalcine) ou enzymatique (ribonucléases, pectate lyases, polygalacturonases). Les structures trimdimensionnelles de la plupart des allergènes polliniques ont été caractérisées, les structures manquantes étant souvent accessibles par modélisation moléculaire à partir de patrons structuraux connus. Les épitopes B liant les IgE d’un petit nombre d’allergènes polliniques ont été identifiés par différentes approches (cartes épitopiques et analyse des complexes allergène-IgE), et les épitopes T de nombreux allergènes sont actuellement connus. Le caractère dégénéré de la reconnaissance des épitopes T par les récepteurs TCR, explique qu’un même TCR puisse reconnaître des épitopes T différents et, inversement, qu’un même épitope T puisse être reconnu par des TCR différents. L’identification de ces épitopes apporte une contribution essentielle dans différents domaines comme l’utilisation des allergènes polliniques recombinants ou naturels pour le diagnostic biologique des pollinoses ou l’utilisation d’épitopes T en immunothérapie spécifique. Ils permettent également de prédire les possibilités de réactions croisées et de co-sensibilisation entre des allergènes polliniques d’origine différente ou dans le syndrome oral entre des allergènes polliniques et des allergènes de fruits ou de légumes. Ces différents aspects moléculaires sont abordés dans cette revue.
Pollen allergens belong to various families of PR (i.e. pathogenesis-related) proteins involved in plant defense mechanisms, such as PR-2 (endo-glucanases), PR-3 (chitinases), PR-5 (thaumatin-like proteins or TLP), PR-10 (Bet v 1-like proteins), PR-12 (defensins) and PR-14 (lipid transfer proteins or LTP). They may also comprise proteins with other functions such as structural function (profilin, expansins), metabolic function (polcalcin) or enzymatic function (ribonucleases, pectate lyases, polygalacturonases) (e.g. ribonucleases, pectate lyases and polygalacturonases). The structural scaffolds of the major weed and tree pollen allergens have been identified either by X-ray crystallography or by NMR for smaller molecules such as LTP and polcalcin. Other pollen allergens of unknown structure have been successfully modelled from templates belonging to the same protein families. Epitope mapping of free allergens and analysis of allergen-IgE complexes enabled identification of IgE-binding B-cell epitopes at the surface of a few pollen allergens. A higher number of T-cell epitopes were identified along the amino acid sequence of various pollen allergens likely to interact with CD4+ T lymphocytes via T-cell receptors (TCRs). However, this low-affinity recognition of T-cell epitopes by TCRs is degenerate, allowing a single TCR to recognize different T-cell epitopes, and, conversely, enabling many different TCRs to recognize the same T-cell epitope. The identification of both B- and T-cell epitopes of pollen allergens is of paramount importance in optimizing the use of natural and recombinant pollen allergens as molecular tools for the component-resolved diagnosis of pollinoses and in improving current immunotherapy practice, for instance by reducing side effects associated with immunotherapy. In addition, identification of B- and T-cell epitopes may provide valuable assistance in the prediction of IgE-binding cross-reactivity and co-sensitization among pollen allergens of different origins, and between pollen and fruit or vegetable allergens responsible for oral allergy syndrome (OAS). Our review aims to present an overview of all these molecular aspects of pollen allergens.
A recombinant isoform of the Ole e 7 olive pollen allergen assembled by de novo mass spectrometry retains the allergenic ability of the natural allergen
2018, Journal of Proteomics
Citation Excerpt :
nsLTPs contain an eight-cysteine residue conserved motif with four disulfide bonds, and an internal hydrophobic cavity where the lipid-binding site is located. Many of these nsLTPs have been described as relevant allergens from different biological sources such as pollen, food or latex [7–15]. Among them, Ole e 7 is a major allergen associated to severe clinical symptoms in those regions with high olive pollen counts, where olive tree is extensively cultivated such as Andalusia in Spain, or Portugal and Italy [3, 16].
The allergenic non-specific lipid transfer protein Ole e 7 from olive pollen is a major allergen associated with severe symptoms in areas with high olive pollen levels. Despite its clinical importance, its cloning and recombinant production has been unable by classical approaches. This study aimed at determining by mass-spectrometry based proteomics its complete amino acid sequence for its subsequent expression and characterization. To this end, the natural protein was in-2D-gel tryptic digested, and CID and HCD fragmentation spectra obtained by nLC-MS/MS analyzed using PEAKS software. Thirteen out of the 457 de novo sequenced peptides obtained allowed assembling its full-length amino acid sequence. Then, Ole e 7-encoding cDNA was synthesized and cloned in pPICZαA vector for its expression in Pichia pastoris yeast. The analyses by Circular Dichroism, and WB, ELISA and cell-based tests using sera and blood from olive pollen-sensitized patients showed that rOle e 7 mostly retained the structural, allergenic and antigenic properties of the natural allergen. In summary, rOle e 7 allergen assembled by de novo peptide sequencing by MS behaved immunologically similar to the natural allergen scarcely isolated from pollen.
Olive pollen is an important cause of allergy. The non-specific lipid binding protein Ole e 7 is a major allergen with a high incidence and a phenotype associated to severe clinical symptoms. Despite its relevance, its cloning and recombinant expression has been unable by classical techniques. Here, we have inferred the primary amino acid sequence of Ole e 7 by mass-spectrometry. We separated Ole e 7 isolated from pollen by 2DE. After in-gel digestion with trypsin and a direct analysis by nLC-MS/MS in an LTQ-Orbitrap Velos, we got the complete de novo sequenced peptides repertoire that allowed the assembling of the primary sequence of Ole e 7. After its protein expression, purification to homogeneity, and structural and immunological characterization using sera from olive pollen allergic patients and cell-based assays, we observed that the recombinant allergen retained the antigenic and allergenic properties of the natural allergen. Collectively, we show that the recombinant protein assembled by proteomics would be suitable for a better in vitro diagnosis of olive pollen allergic patients.
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2016, Allergologia et Immunopathologia
Gramineae bee-collected pollen is identified as being at the origin of allergic accidents but the biological potency of Gramineae bee-collected pollen is not well known. Cereal grasses (e.g., Zea) and European wild forage grasses (FG) are contained in bee-collected pollen.
In this experiment, Zea-mass and FG-mass were identified in bee pollen mass and the proportion of Zea and of FG was calculated using the bee pollen melissopalynology spectrum. Skin reactivity to Zea and to FG were assessed by measuring wheal diameters (W) from skin prick tests using three serial dilutions of bee-collected pollen on 10 allergic patients to Gramineae, in order to calculate the relationship between Zea mass (Mass_zea) or FG mass (Mass_FG) in bee pollen and skin reactivity.
The linear function Log₁₀(W_FG) = 0.24(Log₁₀(Mass_FG)) + 0.33 (R = 0.99) was established using a bee pollen sample with 0.168 mg of FG pollen per mg. The linear function Log₁₀(W_zea) = 0.23(Log₁₀(Mass_zea)) + 0.14 (R = 0.99) was established using a bee pollen sample with 0.983 mg of Zea pollen per mg.
Gramineae allergens seem to be little altered by bee secretions. Gramineae bee pollen retains its allergenic capacity but it depends on the members of the Gramineae family.
To our knowledge this is the first time it has been shown that skin reactivity to Gramineae is proportional to the absolute Gramineae mass contained in the bee-collected pollen and that it depends on the members of the Gramineae family.
The Role of Defensins as Pollen and Food Allergens
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Supported by Instituto de Salud Carlos III (ISCIII-RTIC-G03/094 and RedRespira-ISCIII-RTIC-C03/011), and by Direccion General de Investigacion, Ministerio de Ciencia y Tecnología (project BMC 2002-00196).

Disclosure of potential conflict of interest: The authors have declared they have no conflict of interest.

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Food allergy, dermatologic diseases, and anaphylaxisCabbage lipid transfer protein Bra o 3 is a major allergen responsible for cross-reactivity between plant foods and pollens

Background

Objective

Methods

Results

Conclusion

Clinical implications

Section snippets

Selection of patients and control subjects

General characteristics

Discussion

Ann Allergy Asthma Immunol

Ann Allergy Asthma Immunol

Anal Biochem

J Allergy Clin Immunol

Clin Chim Acta

J Immunol Methods

J Allergy Clin Immunol

Mustard allergy in children

Allergy

Food hypersensitivity in children: clinical aspects and distribution of allergens

Pediatr Allergy Immunol

Contact urticaria from cabbage (Brassica)

Contact Dermatitis

Anaphylaxis to cabbage: detection of allergens

Ann Allergy

Mustard allergy: three cases of systemic reaction to ingestion of mustard sauce

J Investig Allergol Clin Immunol

Anaphylaxis to mustard as a masked allergen in “chicken dips”

Ann Allergy Asthma Immunol

Mustard allergy as a new food allergy

Allergy

Food allergy, dermatologic diseases, and anaphylaxis
Cabbage lipid transfer protein Bra o 3 is a major allergen responsible for cross-reactivity between plant foods and pollens