title: Recombinant Fusion Allergens, Cry j 1 and Cry j 2 from Japanese Cedar Pollen, Conjugated with Polyethylene Glycol Potentiate the Attenuation of Cry j 1-Specific IgE Production in Cry j 1-Sensitized Mice and Japanese Cedar Pollen Allergen-Sensitized Monkeys.
abstract: BACKGROUND
  Japanese cedar (Cryptomeria japonica) pollinosis is the most prevalent seasonal rhinitis in Japan. A standardized Japanese cedar pollen extract (CPE) containing 1.5-4.2 μg of Cry j 1 is currently the highest-concentration extract available for allergen-specific immunotherapy (SIT) against this pollinosis. Therefore, we developed a PEGylated fusion protein as a more effective SIT vaccine against Japanese cedar pollinosis.
  
  
  METHODS
  The fusion protein of major allergens for Japanese cedar pollen, Cry j 1 and Cry j 2, was expressed in Escherichia coli and conjugated with polyethylene glycol (PEG). The purified PEGylated Cry j 1/2 fusion protein (PEG-fusion) was subcutaneously injected four times into Cry j 1- sensitized mice and CPE-sensitized monkeys. The mice were then subcutaneously challenged with Cry j 1 and serum levels of Cry j 1-specific immunoglobulin, and the proliferation and cytokine production of splenocytes were analyzed. The monkeys were intranasally challenged with CPE and analyzed for Cry j 1-specific immunoglobulin levels in plasma.
  
  
  RESULTS
  Cry j 1-specific IgE was significantly attenuated in the PEG-fusion-treated group after Cry j 1-challenge and Cry j 1-specific IgG was significantly increased following PEG-fusion treatment in mice and monkeys. Proliferation and Th2-type cytokine production in splenocytes stimulated with Cry j 1 were also reduced in PEG-fusion-treated mice. IL10 and IL2 production were reduced, but not significantly, while IFN-x03B3; was significantly increased in the PEG-fusion-treated group.
  
  
  CONCLUSIONS
  A high-dose injection of PEG-fusion appears to be a valid candidate for a safer and more effective vaccine than the conventional SIT extract for Japanese cedar pollinosis.
authors.affiliation: RIKEN-TORII Joint Research Team, RIKEN Center for Integrative Medical Sciences (IMS), RIKEN (The Institute of Physical and Chemical Research), Yokohama, Japan.
authors.forename: Takashi
authors.initials: T
authors.lastname: Fujimura
authors.forename: Koji
authors.initials: K
authors.lastname: Fujinami
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Ishikawa
authors.forename: Minoru
authors.initials: M
authors.lastname: Tateno
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Tahara
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Okumura
authors.forename: Hisashi
authors.initials: H
authors.lastname: Ohta
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Miyazaki
authors.forename: Masaru
authors.initials: M
authors.lastname: Taniguchi
mesh_terms: D000485:Allergens / Q000276:immunology*; D000595:Amino Acid Sequence; D000818:Animals; D052179:Antigens, Plant / Q000276:immunology*; D028203:Cedrus / Q000235:genetics*; D049109:Cell Proliferation / Q000502:physiology; D032485:Cryptomeria / Q000276:immunology; D003888:Desensitization, Immunologic / Q000379:methods; D005260:Female; D000882:Haplorhini; D007073:Immunoglobulin E / Q000276:immunology*; D007371:Interferon-gamma / Q000276:immunology; D016753:Interleukin-10 / Q000276:immunology; D007376:Interleukin-2 / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000276:immunology*; D011058:Pollen / Q000276:immunology*; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000276:immunology*
chemical_list: D000485:Allergens; D052179:Antigens, Plant; C065290:Cry j I protein, Cryptomeria japonica; C065291:Cry j II protein, Cryptomeria japonica; D007376:Interleukin-2; D010940:Plant Proteins; D011994:Recombinant Proteins; D016753:Interleukin-10; D007073:Immunoglobulin E; D011092:Polyethylene Glycols; D007371:Interferon-gamma