Takagi Hidenori, Hiroi Takachika, Yang Lijun, Tada Yoshifumi, Yuki Yoshikazu, Takamura Kaoru, Ishimitsu Ryotaro, Kawauchi Hideyuki, Kiyono Hiroshi, Takaiwa Fumio
Plant Biotechnology Department, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki 305-8602, Japan.
Proc Natl Acad Sci U S A. 2005 Nov 29;102(48):17525-30. doi: 10.1073/pnas.0503428102. Epub 2005 Nov 8.
Peptide immunotherapy using multiple predominant allergen-specific T cell epitopes is a safe and promising strategy for the control of type I allergy. In this study, we developed transgenic rice plants expressing mouse dominant T cell epitope peptides of Cry j I and Cry j II allergens of Japanese cedar pollen as a fusion protein with the soybean seed storage protein glycinin. Under the control of the rice seed storage protein glutelin GluB-1 promoter, the fusion protein was specifically expressed and accumulated in seeds at a level of 0.5% of the total seed protein. Oral feeding to mice of transgenic rice seeds expressing the T cell epitope peptides of Cry j I and Cry j II before systemic challenge with total protein of cedar pollen inhibited the development of allergen-specific serum IgE and IgG antibody and CD4(+) T cell proliferative responses. The levels of allergen-specific CD4(+) T cell-derived allergy-associated T helper 2 cytokine production of IL-4, IL-5, and IL-13 and histamine release in serum were significantly decreased. Moreover, the development of pollen-induced clinical symptoms was inhibited in our experimental sneezing mouse model. These results indicate the potential of transgenic rice seeds in production and mucosal delivery of allergen-specific T cell epitope peptides for the induction of oral tolerance to pollen allergens.
使用多种主要过敏原特异性T细胞表位的肽免疫疗法是控制I型过敏的一种安全且有前景的策略。在本研究中,我们培育了转基因水稻植株,其表达日本柳杉花粉的Cry j I和Cry j II过敏原的小鼠显性T细胞表位肽,作为与大豆种子贮藏蛋白大豆球蛋白的融合蛋白。在水稻种子贮藏蛋白谷蛋白GluB-1启动子的控制下,融合蛋白在种子中特异性表达并积累,其水平占种子总蛋白的0.5%。在用柳杉花粉总蛋白进行全身攻击之前,给小鼠口服表达Cry j I和Cry j II的T细胞表位肽的转基因水稻种子,可抑制过敏原特异性血清IgE和IgG抗体以及CD4(+) T细胞增殖反应的产生。过敏原特异性CD4(+) T细胞衍生的与过敏相关的辅助性T细胞2细胞因子IL-4、IL-5和IL-13的产生水平以及血清中的组胺释放均显著降低。此外,在我们的实验性喷嚏小鼠模型中,花粉诱导的临床症状的发展受到抑制。这些结果表明转基因水稻种子在生产和粘膜递送过敏原特异性T细胞表位肽以诱导对花粉过敏原的口服耐受方面具有潜力。