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-γ 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.

Ragweed Pollen Allergy: Burden, Characteristics, and Management of an Imported Allergen Source in Europe

“…Promising results have been attained with these carrier molecules, among which are bacteriophage Qb-derived virus-like particles [121], hepatitis B virus pre-S protein [122-124], and polyethylene glycol [125]. …”

Recombinant Allergens in Structural Biology, Diagnosis, and Immunotherapy

“…Karamloo et al [76 ]could show that a hybrid consisting of fragmented sequences of the 3 bee venom major allergens Api m 1, 2, and 3 had preserved T-cell epitopes, had lost the B cell epitopes of all 3 allergens, and possessed a highly reduced IgE cross-linking ability on human mast cells and basophils as well as a strongly reduced skin test reactivity. The 2 major allergens of Japanese cedar ( Cryptomeria japonica ) pollen, Cry j 1 and Cry j 2, were expressed as a fusion protein and then conjugated to polyethylene glycol to improve their solubility and to create a safer vaccine [77]. Treatment of mice and monkeys with this vaccine resulted in a significant increase of Cry j 1-specific IgG.…”

Next generation immunotherapy for tree pollen allergies

“…To break the Cry j 1 and Cry j 2 conformational structures, the majority of cysteine residues were replaced with serine residues. 88 The modified fusion protein was then conjugated with polyethylene glycol (PEG) and used to treat CPE sensitized monkeys. After four weekly subcutaneous injections of the PEG-conjugated fusion protein administered at a 1mg/0.5ml dose, the production of Cry j 1 specific IgG was significantly increased when compared to the baseline prior to treatment and significantly reduced Cry j 1 specific IgE antibody production.…”