Fujimura Takashi, Fujinami Koji, Ishikawa Ryosuke, Tateno Minoru, Tahara Yoshio, Okumura Yasushi, Ohta Hisashi, Miyazaki Hiroyuki, Taniguchi Masaru
RIKEN-TORII Joint Research Team, RIKEN Center for Integrative Medical Sciences (IMS), RIKEN (The Institute of Physical and Chemical Research), Yokohama, Japan.
Int Arch Allergy Immunol. 2015;168(1):32-43. doi: 10.1159/000441141. Epub 2015 Nov 3.
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.
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.
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.
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.
日本柳杉花粉症是日本最常见的季节性鼻炎。目前，一种含有1.5 - 4.2μg Cry j 1的标准化日本柳杉花粉提取物（CPE）是可用于针对这种花粉症进行变应原特异性免疫治疗（SIT）的最高浓度提取物。因此，我们开发了一种聚乙二醇化融合蛋白，作为一种针对日本柳杉花粉症更有效的SIT疫苗。
日本柳杉花粉主要变应原Cry j 1和Cry j 2的融合蛋白在大肠杆菌中表达，并与聚乙二醇（PEG）偶联。将纯化的聚乙二醇化Cry j 1/2融合蛋白（PEG-融合蛋白）皮下注射到Cry j 1致敏的小鼠和CPE致敏的猴子体内，共注射4次。然后对小鼠进行Cry j 1皮下激发，并分析血清中Cry j 1特异性免疫球蛋白水平以及脾细胞的增殖和细胞因子产生情况。对猴子进行CPE鼻内激发，并分析血浆中Cry j 1特异性免疫球蛋白水平。
在Cry j 1激发后，PEG-融合蛋白治疗组中Cry j 1特异性IgE显著降低，在小鼠和猴子中，PEG-融合蛋白治疗后Cry j 1特异性IgG显著升高。在PEG-融合蛋白治疗的小鼠中，Cry j 1刺激的脾细胞增殖和Th2型细胞因子产生也减少。IL10和IL2的产生减少，但不显著，而在PEG-融合蛋白治疗组中IFN-γ显著增加。
与传统的用于日本柳杉花粉症的SIT提取物相比，高剂量注射PEG-融合蛋白似乎是一种更安全、更有效的疫苗的有效候选物。
