Characterization of the Carbohydrate Moiety in a Purified Allergen Preparation from the Mite <i>Dermatophagoides Farinae</i> and Its Importance for Allergenic Activity as Tested by the RAST-Inhibition Method

Abstract The weed wall pellitory, Parietaria judaica, is one the most important pollen allergen sources in the Mediterranean area causing severe symptoms of hay fever and asthma in allergic patients. We report the expression of the major Parietaria allergens, Par j 1 and Par j 2 which belong to the family of lipid transfer proteins, in insect cells. According to circular dichroism analysis and gel filtration, the purified allergens represented folded and monomeric proteins. Insect cell-expressed, folded Par j 2 exhibited higher IgE binding capacity and more than 100-fold higher allergenic activity than unfolded Escherichia coli-expressed Par j 2 as demonstrated by IgE ELISA and basophil activation testing. IgE ELISA inhibition assays showed that Par j 1 and Par j 2, contain genuine and cross-reactive IgE epitopes. IgG antibodies induced by immunization with Par j 2 inhibited binding of allergic patients IgE to Par j 1 only partially. IgE inhibition experiments demonstrated that insect cell-expressed Par j 1 and Par j 2 together resembled the majority of allergenic epitopes of the Parietaria allergome and therefore both should be used for molecular diagnosis and the design of vaccines for allergen-specific immunotherapy of Parietaria allergy.

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Cloning and Characterization of cDNA Coding for a New Allergen from the House Dust Mite, Dermatophagoides farinae

International Archives of Allergy and Immunology ◽

10.1159/000236653 ◽

1994 ◽

Vol 103 (4) ◽

pp. 349-356 ◽

Cited By ~ 22

Author(s):

Tsunehiro Aki ◽

Kazuhisa Ono ◽

Soon-Young Paik ◽

Takeshi Wada ◽

Toshihiko Jyo ◽

...

Keyword(s):

House Dust ◽

House Dust Mite ◽

Dermatophagoides Farinae ◽

Dust Mite

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Cloning, Expression, and Characterization of Der f 7, an Allergen of Dermatophagoides farinae From China

Journal of Medical Entomology ◽

10.1603/me09180 ◽

2010 ◽

Vol 47 (5) ◽

pp. 868-876 ◽

Cited By ~ 5

Author(s):

Yu-Bao Cui ◽

Hong-Xing Cai ◽

Ying Zhou ◽

Cui-Xiang Gao ◽

Wei-Hong Shi ◽

...

Keyword(s):

Dermatophagoides Farinae

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Purification and characterization of M-177, a 177 kDa allergen, from the house dust mite Dermatophagoides farinae

Allergology International ◽

10.1046/j.1440-1592.1999.00116.x ◽

1999 ◽

Vol 48 (1) ◽

pp. 43-51 ◽

Cited By ~ 3

Author(s):

Akihiro Fujikawa ◽

Seiji Kawamoto ◽

Hideki Hokazono ◽

Tsunehiro Aki ◽

Seiko Shigeta ◽

...

Keyword(s):

House Dust ◽

House Dust Mite ◽

Dermatophagoides Farinae ◽

Purification And Characterization ◽

Dust Mite

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Genomic Organization and Characterization of Group 2 Allergen Paralogs from Dermatophagoides Farinae

Journal of Allergy and Clinical Immunology ◽

10.1016/j.jaci.2005.12.480 ◽

2006 ◽

Vol 117 (2) ◽

pp. S120 ◽

Cited By ~ 1

Author(s):

C.L. Tan ◽

K. Reginald ◽

F.T. Chew

Keyword(s):

Genomic Organization ◽

Dermatophagoides Farinae ◽

Group 2

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Isolation and partial characterization of two antigenic glycoproteins from rye-grass (Lolium perenne) pollen

Biochemical Journal ◽

10.1042/bj1970695 ◽

1981 ◽

Vol 197 (3) ◽

pp. 695-706 ◽

Cited By ~ 25

Author(s):

B J Howlett ◽

A E Clarke

Keyword(s):

Lolium Perenne ◽

Concanavalin A ◽

Sodium Dodecyl Sulphate ◽

Lectin Binding ◽

Sodium Dodecyl ◽

Binding Assays ◽

Pea Lectin ◽

Rye Grass ◽

Allergenic Activity

Two glycoproteins have been purified from a buffer extract of rye-grass (Lolium perenne) pollen. Both migrated as single bands on sodium dodecyl sulphate/polyacrylamide gels. Glycoprotein 1 (0.8 mg/g of pollen) had a apparent mol.wt. of 33 000 and contained 95% protein and 5% carbohydrate. The monosaccharides glucose, galactose, mannose, arabinose and N-acetylglucosamine were present in the proportions 3:3:1:2:1. Glycoprotein 2 (0.4 mg/g of pollen) had an apparent mol. wt. of 68000 and contained 88% protein and 12% carbohydrate. The monosaccharides glucose, galactose, mannose, fucose, xylose, arabinose and N-acetylglucosamine were present in the proportions 4:7:13:5:8:6:6. This glycoprotein bound concanavalin A and Lotus tetragonolobus (asparagus pea) lectin. Radioallergosorbent (RAST) inhibition tests showed that Glycoprotein 1 is an effective allergen, whereas Glycoprotein 2 has less allergenic activity. A method for performing both lectin-binding assays and RAST inhibition tests using microtitre trays is described.

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Hazelnut (Corylus avellana) vicilin Cor a 11: molecular characterization of a glycoprotein and its allergenic activity

Biochemical Journal ◽

10.1042/bj20041062 ◽

2004 ◽

Vol 383 (2) ◽

pp. 327-334 ◽

Cited By ~ 75

Author(s):

Iris LAUER ◽

Kay FOETISCH ◽

Daniel KOLARICH ◽

Barbara K. BALLMER-WEBER ◽

Amedeo CONTI ◽

...

Keyword(s):

Molecular Characterization ◽

Corylus Avellana ◽

Cd Spectroscopy ◽

Mediator Release ◽

Food Allergies ◽

Ige Reactivity ◽

Allergenic Activity ◽

The Impact

In Europe, hazelnuts (Corylus avellana) are a frequent cause of food allergies. Several important hazelnut allergens have been previously identified and characterized. Specific N-glycans are known to induce strong IgE responses of uncertain clinical relevance, but so far the allergenic potential of glycoproteins from hazelnut has not been investigated. The aim of the study was the molecular characterization of the glycosylated vicilin Cor a 11 from hazelnut and the analysis of its allergenic activity. Although MALDI–TOF (matrix-assisted laser-desorption ionization–time-of-flight) MS showed that one of two potential glycosylation sites of Cor a 11 was glycosylated, CD spectroscopy indicated that recombinant and natural Cor a 11 share similar secondary structures. Thus to analyse the impact of the glycan residues of Cor a 11 on IgE binding, the allergenic activity of natural glycosylated Cor a 11 and recombinant Cor a 11 was compared. In addition, the IgE sensitization pattern to recombinant Cor a 11, Cor a 1, Cor a 2 and Cor a 8 of 65 hazelnut allergic patients was determined in vitro. The prevalence of IgE reactivity to hazelnut vicilin Cor a 11 was below 50%. Basophil histamine-release assays were used to determine the allergenic activity of both natural and recombinant Cor a 11 in comparison with Cor a 1, a birch (Betula verrucosa) pollen-related major hazelnut allergen. Both forms of Cor a 11 induced mediator release from basophils to a similar extent, indicating that the hazelnut allergic patients had cross-linking IgE antibodies binding to the protein backbone and not to carbohydrate structures. In comparison to Cor a 1, a 10000-fold higher concentration of Cor a 11 was required to induce similar basophil mediator release. In conclusion, the hazelnut vicilin Cor a 11 is a minor allergen both in regard to prevalence and allergenic potency, whereas its glycan does not contribute to its allergenic activity.

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