title: Identification of wheat sensitization using an in-house wheat extract in Coca-10% alcohol solution in children with wheat anaphylaxis.
abstract: BACKGROUND
  Identification of wheat sensitization by a skin prick test (SPT) is essential for children with wheat-induced anaphylaxis, since oral food challenge can cause serious adverse effects. Wheat allergens are both water/salt and alcohol soluble. The preparation of wheat extract for SPT containing both water/salt and alcohol soluble allergen is needed.
  
  
  OBJECTIVE
  To determine if a wheat extract using Coca's solution containing 10% alcohol (Coca-10% EtOH), prepared in-house, contians both water/salt and alcohol soluble allergens.
  
  
  METHODS
  Serum of children with a history of anaphylaxis after wheat ingestion was used. Wheat flour was extracted in Coca-10% alcohol solution. An SPT with both commercial and in-house wheat extracts was performed as well as specific IgE (sIgE) for wheat and omega-5 gliadin. Direct and IgE inhibition immunoblots were performed to determine serum sIgE levels against water/salt as well as alcohol soluble (gliadins and glutenins) allergens in the extracts.
  
  
  RESULTS
  Six children with history of wheat anaphylaxis had positive SPT to both commercial and in-house extracts. They also had different levels of sIgE against wheat and omega-5 gliadin allergens. The results of direct immunoblotting showed all tested sera had sIgE bound to ~35 kDa wheat protein. Further IgE inhibition immunoblotting identified the ~35 kDa wheat protein as gliadin but not gluten allergen.
  
  
  CONCLUSIONS
  The in-house prepared Coca-10% EtOH solution could extract both water/salt and alcohol soluble allergens. The ~35 kDa gliadin appears to be a major wheat allergen among tested individuals.
authors.affiliation: Division of Allergy and Immunology, Department of Pediatrics, Siriraj Hospital, Mahidol University, Bangkok, Thailand.
authors.forename: Punchama
authors.initials: P
authors.lastname: Pacharn
authors.forename: Sasaros
authors.initials: S
authors.lastname: Kumjim
authors.forename: Puntanat
authors.initials: P
authors.lastname: Tattiyapong
authors.forename: Orathai
authors.initials: O
authors.lastname: Jirapongsananuruk
authors.forename: Surapon
authors.initials: S
authors.lastname: Piboonpocanun
mesh_terms: D000485:Allergens / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology*; D000707:Anaphylaxis / Q000097:blood / Q000139:chemically induced / Q000175:diagnosis* / Q000276:immunology; D015415:Biomarkers / Q000097:blood; D015153:Blotting, Western; D000431:Ethanol / Q000737:chemistry; D005260:Female; D005903:Gliadin / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood; D007223:Infant; D007271:Injections, Intradermal; D008297:Male; D008970:Molecular Weight; D010936:Plant Extracts / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology*; D011237:Predictive Value of Tests; D012882:Skin Tests; D012995:Solubility; D012997:Solvents / Q000737:chemistry; D014908:Triticum / Q000737:chemistry / Q000276:immunology*; D014867:Water / Q000737:chemistry; D021182:Wheat Hypersensitivity / Q000097:blood / Q000175:diagnosis* / Q000276:immunology
chemical_list: D000485:Allergens; D015415:Biomarkers; D010936:Plant Extracts; D012997:Solvents; D014867:Water; D007073:Immunoglobulin E; D000431:Ethanol; D005903:Gliadin