Molecular Characterization of Api g 1, the Major Allergen of Celery (<i>Apium graveolens</i>), and Its Immumological and Structural Relationships to a Group of 17‐kDa Tree Pollen Allergens

Breiteneder, Heimo; Hoffmann‐Sommergruber, Karin; O’Riordain, Gabriel; Susani, Markus; Ahorn, Horst; Ebner, Christof; Kraft, Dietrich; Scheiner, Otto

doi:10.1111/j.1432-1033.1995.484_2.x

Cited by 227 publications

(161 citation statements)

References 32 publications

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“…Recombinant allergens, Bet v 1-derived peptides, rabbit Abs, and sera from allergic patients Recombinant birch pollen allergen Bet v 1a, recombinant hypoallergenic isoform Bet v 1d, and Bet v 1-related recombinant allergens Aln g 1, Cor a 1, Mal d 1, Api g 1, and Dau c 1 were obtained from Biomay (Vienna, Austria) (27)(28)(29)(30)(31)(32)(33). Bet v 1-derived peptides (Table I) were synthesized and purified as described (26).…”

Section: Methodsmentioning

confidence: 99%

Mapping of Conformational IgE Epitopes with Peptide-Specific Monoclonal Antibodies Reveals Simultaneous Binding of Different IgE Antibodies to a Surface Patch on the Major Birch Pollen Allergen, Bet v 1

Gieras

Cejka

Blatt

et al. 2011

The Journal of Immunology

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Allergic inflammation is based on the cross-linking of mast cell and basophil-bound IgE Abs and requires at least two binding sites for IgE on allergens, which are difficult to characterize because they are often conformational in nature. We studied the IgE recognition of birch pollen allergen Bet v 1, a major allergen for >100 million allergic patients. Monoclonal and polyclonal Abs raised against Bet v 1-derived peptides were used to compete with allergic patients’ IgE binding to Bet v 1 to search for sequences involved in IgE recognition. Strong inhibitions of patients’ IgE binding to Bet v 1 (52–75%) were obtained with mAbs specific for two peptides comprising aa 29–58 (P2) and aa 73–103 (P6) of Bet v 1. As determined by surface plasmon resonance, mAb2 specific for P2 and mAb12 specific for P6 showed high affinity, but only polyclonal rabbit anti-P2 and anti-P6 Abs or a combination of mAbs inhibited allergen-induced basophil degranulation. Thus, P2 and P6 define a surface patch on the Bet v 1 allergen, which allows simultaneous binding of several different IgE Abs required for efficient basophil and mast cell activation. This finding explains the high allergenic activity of the Bet v 1 allergen. The approach of using peptide-specific Abs for the mapping of conformational IgE epitopes on allergens may be generally applicable. It may allow discriminating highly allergenic from less allergenic allergen molecules and facilitate the rational design of active and passive allergen-specific immunotherapy strategies.

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Section: Methodsmentioning

confidence: 99%

Mapping of Conformational IgE Epitopes with Peptide-Specific Monoclonal Antibodies Reveals Simultaneous Binding of Different IgE Antibodies to a Surface Patch on the Major Birch Pollen Allergen, Bet v 1

Gieras

Cejka

Blatt

et al. 2011

The Journal of Immunology

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“…This observation clearly differed from other Bet v 1‐related food allergens, in particular from Api g 1 in celery, which has an aa sequence similarity of 93% (81% identity) with Dau c 1 7. In contrast to Dau c 1, IgE binding to Api g 1 is completely abolished by Bet v 1 10, 11. Additional evidence for a Bet v 1‐independent Dau c 1‐specific IgE response has been provided by the detection of significant Dau c 1‐specific IgE levels in 4 adult patients with strong immediate systemic reactions to raw carrot who displayed no Bet v 1‐specific IgE antibodies 12.…”

mentioning

confidence: 66%

Characterization of the T-cell response to Dau c 1, the Bet v 1-homolog in carrot

Zulehner

Nagl

Briza

et al. 2016

Allergy

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BackgroundIn contrast to other Bet v 1‐related food allergens, the major carrot allergen, Dau c 1, has been suggested to induce food allergy independently from Bet v 1. As T cells are crucial in the sensitization process, we sought to characterize the T‐cell response to Dau c 1 and its cross‐reactivity with Bet v 1.MethodsDau c 1‐specific T‐cell lines (TCL) and clones (TCC) established from PBMC of birch pollen‐allergic patients with carrot allergy were analyzed for reactivity to Bet v 1, epitope specificity, allergen‐induced cytokine secretion, and expression of integrins α4β7 and α4β1, critical for gut and lung homing, respectively. mRNA expression of GATA3 and Tbet was analyzed in sorted CD3+ CD4+ CFSE low cells proliferating upon stimulation of PBMC with Dau c 1 or Bet v 1. Dau c 1 was incubated with endolysosomal proteases, and the resulting fragments were identified by mass spectrometry.ResultsAmong 14 distinct T‐cell‐activating regions, Dau c 1139–153 was recognized by 55% of the patients. Only 6 of 15 (40%) Dau c 1‐specific TCL and 9 of 21 (43%) TCC reacted with Bet v 1. Bet v 1‐nonreactive TCC were mainly Th1‐like and showed a higher expression of the integrin β7 and a significantly lower expression of the integrin β1 than Bet v 1‐positive TCC. A Th1‐like response was also detected in Dau c 1‐reactive CD3+ CD4+ CFSE low cells. Full‐length Dau c 1 was still detectable after 48 h of endolysosomal degradation. Proteolytic fragments of Dau c 1 matched its T‐cell‐activating regions.ConclusionDau c 1 displays several characteristics of sensitizing allergens, namely a major T‐cell‐activating region, low susceptibility to endolysosomal degradation, and induction of a Bet v 1‐independent T‐cell response. These cellular insights confirm that the major carrot allergen has a special status among Bet v 1‐related food allergens.

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“…After full length cDNA cloning and expression, this hypothesis will be tested by enzymatic assays using the specific substrates of IFR. Bet v 1 and its related food allergens are thought to belong to a group of intracellular pathogenesis-related proteins being involved in plant defence mechanisms [5,20]. Our ongoing investigation will highlight whether another group of 'defence proteins' creates important allergens in pollen and food.…”

Section: Discussionmentioning

confidence: 99%

“…These are due to homologues of Bet v 1, the major birch pollen allergen, which are present in many plants [3,4]. Examples of cross-reactive major food allergens belonging to this family are Mal d 1, Pru a 1 and Api g 1, from apples, cherries and celery tubers, respectively [5][6][7][8]. All these proteins have molecular masses (MM) between 16 kDa and 18 kDa.…”

Section: Introductionmentioning

confidence: 99%

Characterization of a New IgE‐Binding 35‐kDa Protein from Birch Pollen with Cross‐Reacting Homologues in Various Plant Foods

et al. 1998

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The present investigation was undertaken to obtain molecular data of a new immunoglobulin (Ig)E-binding birch pollen protein with a mass of 35 kDa. In a previous study, this protein showed IgE cross-reactivity with 34-and 35-kDa proteins in apples, pears, carrots, bananas and other exotic fruits. Since the protein was Nterminally blocked, it was purified by preparative SDS-PAGE, and multiple proteolytic fragments were subsequently generated by in-gel digestion with the endoproteinases Glu C, Lys C and Clostripain. After electrophoretic separation and blotting onto polyvinylidene difluoride (PVDF), the resulting polypeptides were subjected to N-terminal amino acid microsequencing. The internal sequences obtained showed a high degree of sequence identity to isoflavone reductases (IFR) and isoflavone reductase-like proteins (IRL) from several plants which also had a similar size. For a stretch of 25 consecutive residues this identity ranged from 56% for IFR from peas and chick peas and an IRL from maize, to 80% for a tobacco IRL. A 453 bp fragment was amplified from total birch pollen RNA by polymerase chain reaction (PCR) using primers derived from the nucleotide sequence of the tobacco IRL. The deduced 151 amino acid sequence represented Ϸ 50% of the protein and confirmed the sequence identities obtained by Edman degradation. Moreover, the 25 amino acid sequence was included in the cloned fragment. Deduced and determined amino acids showed only one mismatch, which was due to a single nucleotide exchange. At the antibody level, the immunological relationship of the birch pollen protein to IRL and IFR was demonstrated by immunoblotting with a rabbit antiserum against a pea IFR which recognized the same birch protein as patients' IgE. The rabbit antiserum also reproduced the cross-reactivity pattern previously observed with patients' IgE by recognizing related proteins in specific plant foods, including some exotic fruits. We therefore suggest that the 35-kDa birch pollen protein belongs to the IFR/IRL family and represents a minor allergen, possibly being responsible for less common pollen-related food allergies in patients allergic to birch pollen.

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Molecular Characterization of Api g 1, the Major Allergen of Celery (Apium graveolens), and Its Immumological and Structural Relationships to a Group of 17‐kDa Tree Pollen Allergens

Cited by 227 publications

References 32 publications

Mapping of Conformational IgE Epitopes with Peptide-Specific Monoclonal Antibodies Reveals Simultaneous Binding of Different IgE Antibodies to a Surface Patch on the Major Birch Pollen Allergen, Bet v 1

Mapping of Conformational IgE Epitopes with Peptide-Specific Monoclonal Antibodies Reveals Simultaneous Binding of Different IgE Antibodies to a Surface Patch on the Major Birch Pollen Allergen, Bet v 1

Characterization of the T-cell response to Dau c 1, the Bet v 1-homolog in carrot

Characterization of a New IgE‐Binding 35‐kDa Protein from Birch Pollen with Cross‐Reacting Homologues in Various Plant Foods

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