title: Recombinant Pru p 3 and natural Pru p 3, a major peach allergen, show equivalent immunologic reactivity: a new tool for the diagnosis of fruit allergy.
abstract: BACKGROUND
  The peach lipid transfer protein Pru p 3 has been identified as a major allergen from this fruit. Homologous cross-reactive allergens have been found in several plant foods and pollens. Recombinant Pru p 3 has been recently produced in the yeast Pichia pastoris.
  
  
  OBJECTIVE
  We sought to evaluate the potential role of recombinant Pru p 3 as a novel tool for the diagnosis of fruit allergy.
  
  
  METHODS
  Circular dichroism analysis was used to compare the protein folding of natural Pru p 3 and recombinant Pru p 3. IgE binding by both molecular forms was quantified by means of ELISA and ELISA inhibition assays, and their biologic activity was estimated by using basophil activation, histamine release, and sulphidoleukotriene production tests. Individual sera or blood samples from patients with peach allergy (up to 17) were used in the assays.
  
  
  RESULTS
  A nearly identical circular dichroism spectra was shown by using natural Pru p 3 and recombinant Pru p 3, indicating that both protein forms are similarly folded. No difference was detected in the IgE-binding capacity of the 2 mo-lecular versions. Basophil activation and induction of sulphidoleukotriene production were positive in 9 of 10 patients, and histamine release was induced in at least half of the patients, with similar effects of the natural and recombinant forms in the 3 assays.
  
  
  CONCLUSIONS
  Recombinant Pru p 3 shows a strong immunologic activity equivalent to that of its natural counterpart, and therefore it can be a useful tool for diagnosis (and future immunotherapy) of fruit allergy.
authors.affiliation: Unidad de Bioquímica, Departamento de Biotecnología, E.T.S. Ingenieros Agrónomos, Madrid, Spain.
authors.forename: Araceli
authors.initials: A
authors.lastname: Díaz-Perales
authors.forename: María L
authors.initials: ML
authors.lastname: Sanz
authors.forename: Gloria
authors.initials: G
authors.lastname: García-Casado
authors.forename: Rosa
authors.initials: R
authors.lastname: Sánchez-Monge
authors.forename: Francisco J
authors.initials: FJ
authors.lastname: García-Selles
authors.forename: Manuel
authors.initials: M
authors.lastname: Lombardero
authors.forename: Florentino
authors.initials: F
authors.lastname: Polo
authors.forename: Pedro M
authors.initials: PM
authors.lastname: Gamboa
authors.forename: Domingo
authors.initials: D
authors.lastname: Barber
authors.forename: Gabriel
authors.initials: G
authors.lastname: Salcedo
mesh_terms: D000293:Adolescent; D000328:Adult; D000485:Allergens / Q000276:immunology*; D052179:Antigens, Plant; D001491:Basophils / Q000502:physiology; D002942:Circular Dichroism; D005260:Female; D005512:Food Hypersensitivity / Q000175:diagnosis*; D005638:Fruit / Q000009:adverse effects*; D006636:Histamine Release; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology; D015289:Leukotrienes / Q000096:biosynthesis; D008297:Male; D010940:Plant Proteins; D017510:Protein Folding; D011994:Recombinant Proteins / Q000276:immunology
chemical_list: D000485:Allergens; D052179:Antigens, Plant; D015289:Leukotrienes; D010940:Plant Proteins; C463181:Pru p 3 allergen; D011994:Recombinant Proteins; D007073:Immunoglobulin E