Profilin (Che a 2) and polcalcin (Che a 3) are relevant allergens of Chenopodium album pollen: Isolation, amino acid sequences, and immunologic properties

Barderas, Rodrigo; Villalba, Mayte; Pascual, Cristina; Batanero, Eva; Rodrı́guez, Rosalı́a

doi:10.1016/j.jaci.2003.12.587

Cited by 67 publications

(59 citation statements)

References 32 publications

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“…Patients were selected because of a well-known sensitivity to chenopod pollen [12, 14]. All patients and control individuals provided informed consent.…”

Section: Methodsmentioning

confidence: 99%

“…Recently, our group has reported the existence of Che a 1, Che a 2 (profilin) and Che a 3 (polcalcin) as relevant allergens in C. album pollen [12, 13, 14], weed that defines the name of the Chenopodiaceae family. Che a 1 and Che a 2 have been described as major allergens, since these proteins are recognized by the IgE from at least 50% of patients suffering from chenopod sensitization.…”

Section: Introductionmentioning

confidence: 99%

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Che a 1: Recombinant Expression, Purification and Correspondence to the Natural Form

Barderas¹,

Villalba²,

Rodrı́guez³

2004

Int Arch Allergy Immunol

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Background: Pollinosis to chenopods is one of the main causes of allergy in desertic regions and it is increasing in the South of Europe and Western USA. Che a 1 is a major allergen for chenopod-allergic subjects and belongs to the Ole-e-1-like family of proteins. Methods:Pichia pastoris yeast has been used as expression system to produce the recombinant form of Che a 1 (rChe a 1). The allergen was isolated using a gel permeation column and reverse-phase/high-performance liquid chromatography. Molecular characterization was performed using Edman degradation, mass spectrometry and concanavalin A staining. Sera from patients allergic to chenopod pollen, as well as polyclonal and monoclonal antibodies raised against Ole e 1, were used in immunoblotting, ELISA and inhibition assays for immunological characterization of rChe a 1. Results: The allergen was purified to homogeneity with a final yield of 15 mg/l of cell culture and showed a glycosylated character. N-terminal amino acid sequence of rChe a 1 and molecular mass were according to those of the protein isolated from chenopod pollen. The recombinant allergen maintained the IgG and IgE epitopes of the natural allergen deduced from the immunological assays. Conclusions: Structural and in vitro immunological properties of rChe a 1 produced in P. pastoris were equivalent to those of the natural form of the allergen and, thus, it could be used in testing patients allergic to chenopods.

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“…Patients were selected because of a well-known sensitivity to chenopod pollen [12, 14]. All patients and control individuals provided informed consent.…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Che a 1: Recombinant Expression, Purification and Correspondence to the Natural Form

Barderas¹,

Villalba²,

Rodrı́guez³

2004

Int Arch Allergy Immunol

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“…In another study, IgE binding to profilin from sunflower pollen was also about 30% in a population of sunflower-allergic patients from sunflower-growing areas [26]. Profilin seems to give rise to a higher frequency of sensitization among patients allergic to Chenopodium album [27], Mercurialis annua [28] and Phoenix dactylifera [29] pollen, but in some of these studies sensitization to profilin was only determined by IgE binding on immunoblot at the level of profilin’s molecular mass. Therefore, possibly other allergens with similar apparent mass could be involved and the potential role of profilin as a major allergen in these pollens should therefore still be confirmed using natural purified or recombinant profilins from the respective sources.…”

Section: Profilin: Prevalence Of Sensitizationmentioning

confidence: 99%

Profilins: Mimickers of Allergy or Relevant Allergens?

Santos

Ree

2011

Int Arch Allergy Immunol

145

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Profilins are ubiquitous proteins, present in all eukaryotic cells and identified as allergens in pollen, latex and plant foods. The highly conserved structure justifies the cross-reactive nature of IgE antibodies against plant profilins and their designation as pan-allergens. Primary sensitization to profilin seems to arise from pollen sensitization with later development of cross-reactive IgE antibodies against plant food (and possibly latex) profilins. The role of profilin in inducing allergic symptoms needs to be evaluated and raises important issues in allergy diagnosis due to cross-reactivity. IgE cross-reactivity among profilins is associated with multiple pollen sensitization and with various pollen-food syndromes. In respiratory allergy, sensitization to pollen to which the patient has virtually no environmental exposure has been identified as a manifestation of profilin sensitization. As a food allergen, profilin usually elicits mild reactions, such as oral allergy syndrome, is not modified by processing and is especially important in allergy to some fruits, such as melon, watermelon, banana, tomato, citrus fruit and persimmon. Purified natural and recombinant profilins for in vitro and in vivo allergy tests are helpful in the diagnostic work-up. Herein we review the current state of knowledge about the allergen profilin and its implications in the diagnosis and treatment of allergic diseases. We conclude that, although its role in triggering allergic symptoms is still controversial, profilin is undoubtedly a relevant allergen. As a pan-allergen, profilin is associated with multiple pollen sensitization and pollen-food-latex syndromes that the allergist has to be aware of in order to accomplish an accurate diagnosis and successful treatment of allergic diseases.

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“…16) was subcloned into the NdeI/ EcoRI restriction sites of the pET11b expression plasmid essentially as described earlier (17). Overnight cultures of Escherichia coli BL21(DE3) transformed with the pET11b/Che a 3 construct were diluted 10-fold with Luria-Bertani medium containing 0.1 mg/ml ampicillin and grown to an OD 600 nm of 1.0.…”

Section: Cloning Expression and Purification Of Recombinant Che Amentioning

confidence: 99%

“…Sera were obtained from Austrian and Spanish patients with polysensitization to pollens from various unrelated plants (e.g., trees, grasses, and weeds) and tested for the presence of IgE Abs specific for two EF-hand calcium-binding allergens (16,33). Rabbit Abs and the mouse mAb raised against the two EF-hand allergen from olive pollen, Ole e 3, have been described elsewhere (17).…”

Section: Abs Sera and Immunological Assaysmentioning

confidence: 99%

Three-Dimensional Structure of the Cross-Reactive Pollen Allergen Che a 3: Visualizing Cross-Reactivity on the Molecular Surfaces of Weed, Grass, and Tree Pollen Allergens

Verdino

Barderas

Villalba

et al. 2008

The Journal of Immunology

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Two EF-hand calcium-binding allergens (polcalcins) occur in the pollen of a wide variety of unrelated plants as highly cross-reactive allergenic molecules. We report the expression, purification, immunological characterization, and the 1.75-Å crystal structure of recombinant Che a 3 (rChe a 3), the polcalcin from the weed Chenopodium album. The three-dimensional structure of rChe a 3 resembles an α-helical fold that is essentially identical with that of the two EF-hand allergens from birch pollen, Bet v 4, and timothy grass pollen, Phl p 7. The extensive cross-reactivity between Che a 3 and Phl p 7 is demonstrated by competition experiments with IgE Abs from allergic patients as well as specific Ab probes. Amino acid residues that are conserved for the two EF-hand allergen family were identified in multiple sequence alignments of polcalcins from 15 different plants. Next, the three-dimensional structures of rChe a 3, rPhl p 7, and rBet v 4 were used to identify conserved amino acids with high surface exposition to visualize surface patches as potential targets for the polyclonal IgE Ab response of allergic patients. The essentially identical three-dimensional structures of rChe a 3, rPhl p 7, and rBet v 4 explain the extensive cross-reactivity of allergic patients IgE Abs with two EF-hand allergens from unrelated plants. In addition, analyzing the three-dimensional structures of cross-reactive Ags for conserved and surface exposed amino acids may be a first approach to mapping the conformational epitopes on disease-related Ags that are recognized by polyclonal patient Abs.

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Profilin (Che a 2) and polcalcin (Che a 3) are relevant allergens of Chenopodium album pollen: Isolation, amino acid sequences, and immunologic properties

Cited by 67 publications

References 32 publications

Che a 1: Recombinant Expression, Purification and Correspondence to the Natural Form

Che a 1: Recombinant Expression, Purification and Correspondence to the Natural Form

Profilins: Mimickers of Allergy or Relevant Allergens?

Three-Dimensional Structure of the Cross-Reactive Pollen Allergen Che a 3: Visualizing Cross-Reactivity on the Molecular Surfaces of Weed, Grass, and Tree Pollen Allergens

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