Recombinant allergens for diagnosis and therapy of allergic disease

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Presentation transcript:

Recombinant allergens for diagnosis and therapy of allergic disease Martin D. Chapman, PhDa, Alisa M. Smith, PhDa, Lisa D. Vailes, MSa, L.Karla Arruda, MD, PhDb, Venugopal Dhanaraj, PhDc, Anna Pomés, PhDa Journal of Allergy and Clinical Immunology Volume 106, Issue 3, Pages 409-418 (September 2000) DOI: 10.1067/mai.2000.109832 Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 1 Tertiary structure of a lipocalin allergen: three spatial perspective views of rodent urinary allergen, Rat n 1 (chain A from 2A2U.pdb).26 Spheres represent amino acids that form sequence motifs located in the three structurally conserved regions of lipocalins.25 These residues are located at the base of the cup region of the molecule, which forms the ligand-binding pocket. Journal of Allergy and Clinical Immunology 2000 106, 409-418DOI: (10.1067/mai.2000.109832) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 2 Comparison of the three-dimensional structure of an active aspartic protease (porcine pepsin) with the inactive cockroach aspartic protease allergen Bla g 2.13,21,48,49 The active site of pepsin contains two identical catalytic triads (amino acid residues DTG) with the coplanar aspartate residues at positions 32 (in yellow) and 215 (green). In Bla g 2, the triad sequences are DST and DTS, which results in a loss of catalytic activity. The substitution of the tyrosine residue at position 75 for phenylalanine in Bla g 2 also contributes to the loss of aspartic protease activity in Bla g 2.13,21 Journal of Allergy and Clinical Immunology 2000 106, 409-418DOI: (10.1067/mai.2000.109832) Copyright © 2000 Mosby, Inc. Terms and Conditions

Fig. 3 Immediate intradermal skin tests to recombinant (r) Blo t 5 and Der p 5. This patient had positive prick test result to 5 μg/mL recombinant Blo t 5 and to recombinant Der p 5 (not shown) and was given intradermal tests with serial 10-fold dilutions of the allergens to assess their potency. The patient had 8 × 8 mm wheals to 10–3 μg/mL each allergen, similar to B tropicalis or D pteronyssinus extract. Note that the potency of the recombinants can be directly compared in micrograms per milliliter, whereas the potency of the allergen extracts cannot (these are expressed in micrograms per milliliter of total protein or in arbitrary allergy units per milliliter). The patient was also prick tested with glutathione-S-transferase (GST) expression vector sequence as a negative control. Journal of Allergy and Clinical Immunology 2000 106, 409-418DOI: (10.1067/mai.2000.109832) Copyright © 2000 Mosby, Inc. Terms and Conditions