title: Comparison of inflammatory responses to genetically engineered hypoallergenic derivatives of the major birch pollen allergen bet v 1 and to recombinant bet v 1 wild type in skin chamber fluids collected from birch pollen-allergic patients.
abstract: BACKGROUND
  Nearly 60% of birch pollen-allergic patients react exclusively to Bet v 1. With use of the skin blister model, previously only established for installation of crude allergens, we have for the first time characterized the inflammatory response in vivo to recombinant birch pollen allergen, rBet v 1, molecules (rBet v 1 wild type, fragments and trimer).
  
  
  OBJECTIVE
  Our purpose was to examine whether challenge with rBet v 1 derivatives (fragments and trimer) compared with rBet v 1 wild type differs with respect to influx of activated eosinophils and detectable levels of cytokines/chemokines related to allergic inflammation in skin chambers applied to birch pollen-allergic patients.
  
  
  METHODS
  The skin blister chambers were filled for 2 hours with rBet v 1, the derivatives or PBS and heparin (negative control). The fluids were analyzed after 2 and 8 hours. The number of eosinophils was determined and EG2 and CD69 expression measured by flow cytometry. Cytokines and mediators were analyzed by ELISA and RIA techniques.
  
  
  RESULTS
  Comparable numbers of eosinophils were recruited to the chambers challenged with rBet v 1 molecules, but the eosinophils from the rBet v 1 wild-type challenged chambers showed a significantly higher expression of CD69. The levels of eotaxin were similar in all 4 chambers, whereas rBet v 1 wild type induced significantly higher levels of histamine, eosinophil cationic protein, and GM-CSF than the derivatives did. Recombinant Bet v 1 trimer elicited significantly lower levels of IL-4 compared with rBet v1 wild type.
  
  
  CONCLUSIONS
  Genetically engineered hypoallergenic rBet v 1 derivatives recruited eosinophils analogously with rBet v 1 wild type. However, the derivatives exhibited a lower capacity to activate eosinophils and to release proinflammatory mediators and T helper type 2-derived cytokines. The derivatives may therefore be candidate molecules for specific immunotherapy of birch pollen allergy with reduced risk of inducing allergenic or inflammatory side effects.
authors.affiliation: Division of Clinical Immunology, Karolinska Hospital and Institutet, Stockholm, Sweden.
authors.forename: A
authors.initials: A
authors.lastname: Nopp
authors.forename: G
authors.initials: G
authors.lastname: Halldén
authors.forename: J
authors.initials: J
authors.lastname: Lundahl
authors.forename: E
authors.initials: E
authors.lastname: Johansson
authors.forename: S
authors.initials: S
authors.lastname: Vrtala
authors.forename: R
authors.initials: R
authors.lastname: Valenta
authors.forename: R
authors.initials: R
authors.lastname: Grönneberg
authors.forename: M
authors.initials: M
authors.lastname: Van Hage-Hamsten
mesh_terms: D000293:Adolescent; D000328:Adult; D000485:Allergens; D015703:Antigens, CD / Q000032:analysis; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis; D052179:Antigens, Plant; D001798:Blood Proteins / Q000032:analysis; D054413:Chemokine CCL11; D019742:Chemokines, CC; D016207:Cytokines / Q000032:analysis; D047091:Eosinophil Granule Proteins; D005260:Female; D005818:Genetic Engineering; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000032:analysis; D006801:Humans; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000032:analysis; D015847:Interleukin-4 / Q000032:analysis; D037181:Lectins, C-Type; D008297:Male; D008875:Middle Aged; D010940:Plant Proteins / Q000235:genetics* / Q000276:immunology*; D011058:Pollen / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D012260:Ribonucleases; D012867:Skin / Q000276:immunology*; D014197:Trees; D000099192:CD69 Antigens
chemical_list: D000485:Allergens; D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D052179:Antigens, Plant; D001798:Blood Proteins; D054413:Chemokine CCL11; D019742:Chemokines, CC; D016207:Cytokines; D047091:Eosinophil Granule Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D007371:Interferon-gamma; D015847:Interleukin-4; D037181:Lectins, C-Type; D010940:Plant Proteins; D011994:Recombinant Proteins; D012260:Ribonucleases; D000099192:CD69 Antigens; C515456:CCL11 protein, human; C061312:Bet v 1 allergen, Betula