title: Development and characterization of a recombinant, hypoallergenic, peptide-based vaccine for grass pollen allergy.
abstract: BACKGROUND
  Grass pollen is one of the most important sources of respiratory allergies worldwide.
  
  
  OBJECTIVE
  This study describes the development of a grass pollen allergy vaccine based on recombinant hypoallergenic derivatives of the major timothy grass pollen allergens Phl p 1, Phl p 2, Phl p 5, and Phl p 6 by using a peptide-carrier approach.
  
  
  METHODS
  Fusion proteins consisting of nonallergenic peptides from the 4 major timothy grass pollen allergens and the PreS protein from hepatitis B virus as a carrier were expressed in Escherichia coli and purified by means of chromatography. Recombinant PreS fusion proteins were tested for allergenic activity and T-cell activation by means of IgE serology, basophil activation testing, T-cell proliferation assays, and xMAP Luminex technology in patients with grass pollen allergy. Rabbits were immunized with PreS fusion proteins to characterize their immunogenicity.
  
  
  RESULTS
  Ten hypoallergenic PreS fusion proteins were constructed, expressed, and purified. According to immunogenicity and induction of allergen-specific blocking IgG antibodies, 4 hypoallergenic fusion proteins (BM321, BM322, BM325, and BM326) representing Phl p 1, Phl p 2, Phl p 5, and Phl p 6 were included as components in the vaccine termed BM32. BM321, BM322, BM325, and BM326 showed almost completely abolished allergenic activity and induced significantly reduced T-cell proliferation and release of proinflammatory cytokines in patients' PBMCs compared with grass pollen allergens. On immunization, they induced allergen-specific IgG antibodies, which inhibited patients' IgE binding to all 4 major allergens of grass pollen, as well as allergen-induced basophil activation.
  
  
  CONCLUSIONS
  A recombinant hypoallergenic grass pollen allergy vaccine (BM32) consisting of 4 recombinant PreS-fused grass pollen allergen peptides was developed for safe immunotherapy of grass pollen allergy.
authors.affiliation: Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria.
authors.forename: Margarete
authors.initials: M
authors.lastname: Focke-Tejkl
authors.affiliation: Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria.
authors.forename: Milena
authors.initials: M
authors.lastname: Weber
authors.affiliation: Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Niespodziana
authors.affiliation: Biomay AG, Vienna, Austria.
authors.forename: Angela
authors.initials: A
authors.lastname: Neubauer
authors.affiliation: Biomay AG, Vienna, Austria.
authors.forename: Hans
authors.initials: H
authors.lastname: Huber
authors.affiliation: Biomay AG, Vienna, Austria.
authors.forename: Rainer
authors.initials: R
authors.lastname: Henning
authors.affiliation: Biomay AG, Vienna, Austria.
authors.forename: Gottfried
authors.initials: G
authors.lastname: Stegfellner
authors.affiliation: Biomay AG, Vienna, Austria.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Maderegger
authors.affiliation: Biomay AG, Vienna, Austria.
authors.forename: Martina
authors.initials: M
authors.lastname: Hauer
authors.affiliation: Biomay AG, Vienna, Austria.
authors.forename: Frank
authors.initials: F
authors.lastname: Stolz
authors.affiliation: Department of Otorhinolaryngology, Medical University of Vienna, Vienna, Austria.
authors.forename: Verena
authors.initials: V
authors.lastname: Niederberger
authors.affiliation: Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria.
authors.forename: Katharina
authors.initials: K
authors.lastname: Marth
authors.affiliation: Department of Otorhinolaryngology, Medical University of Vienna, Vienna, Austria.
authors.forename: Julia
authors.initials: J
authors.lastname: Eckl-Dorna
authors.affiliation: Department of Molecular Biology, Division of Allergy and Immunology, University of Salzburg, Salzburg, Austria.
authors.forename: Richard
authors.initials: R
authors.lastname: Weiss
authors.affiliation: Department of Molecular Biology, Division of Allergy and Immunology, University of Salzburg, Salzburg, Austria.
authors.forename: Josef
authors.initials: J
authors.lastname: Thalhamer
authors.affiliation: Department of Internal Medicine I, Division of Hematology & Hemostaseology, Medical University of Vienna, Vienna, Austria.
authors.forename: Katharina
authors.initials: K
authors.lastname: Blatt
authors.affiliation: Department of Internal Medicine I, Division of Hematology & Hemostaseology, Medical University of Vienna, Vienna, Austria.
authors.forename: Peter
authors.initials: P
authors.lastname: Valent
authors.affiliation: Division of Immunopathology, Department of Pathophysiology and Allergy Research, Center of Pathophysiology, Infectiology and Immunology, Medical University of Vienna, Vienna, Austria. Electronic address: rudolf.valenta@meduniwien.ac.at.
authors.forename: Rudolf
authors.initials: R
authors.lastname: Valenta
mesh_terms: D000485:Allergens / Q000276:immunology; D000818:Animals; D001491:Basophils / Q000276:immunology / Q000378:metabolism; D016207:Cytokines / Q000096:biosynthesis; D004195:Disease Models, Animal; D005260:Female; D006514:Hepatitis B Surface Antigens / Q000737:chemistry / Q000235:genetics; D006801:Humans; D007073:Immunoglobulin E / Q000097:blood / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology; D018836:Inflammation Mediators / Q000378:metabolism; D008213:Lymphocyte Activation / Q000276:immunology; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism; D051379:Mice; D010455:Peptides / Q000276:immunology; D006109:Poaceae; D011058:Pollen / Q000276:immunology; D011485:Protein Binding; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D006255:Rhinitis, Allergic, Seasonal / Q000517:prevention & control*; D022223:Vaccines, Subunit / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000276:immunology*
chemical_list: D000485:Allergens; D016207:Cytokines; D006514:Hepatitis B Surface Antigens; D007074:Immunoglobulin G; D018836:Inflammation Mediators; D010455:Peptides; D011993:Recombinant Fusion Proteins; D022223:Vaccines, Subunit; D014614:Vaccines, Synthetic; D007073:Immunoglobulin E
keywords: Grass pollen allergy; allergen; allergen-specific immunotherapy; peptide-carrier technology; recombinant allergen; recombinant hypoallergenic allergen derivative