Fujimura Takashi, Shigeta Seiko, Kawamoto Seiji, Aki Tsunehiro, Masubuchi Masanori, Hayashi Takaharu, Yoshizato Katsutoshi, Ono Kazuhisa
Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, Higashi-Hiroshima, Japan.
Int Arch Allergy Immunol. 2004 Feb;133(2):125-35. doi: 10.1159/000076438. Epub 2004 Jan 26.
Japanese cedar (Cryptomeria japonica) pollen is one of the most prevalent sources of the allergens that elicit rhinitis and conjunctivitis. Only Cry j 1 and Cry j 2 have been well characterized as the major allergens of this pollen.
The aims of this study were to complete the repertoire of C. japonica pollen allergens, to investigate their variability with respect to IgE-reactive patterns and to identify the isoforms of Cry j 1 and Cry j 2 by proteome analysis.
Proteins in C. japonica pollen separated on two-dimensional (2-D) polyacrylamide gel electrophoresis were immunodetected with IgE in sera of 40 subjects allergic to C. japonica pollen. Mass fingerprinting was used to elucidate the diversity of the major allergens.
2-D immunolabeling with individual patients' sera showed the distinguishable IgE-binding patterns inlaid with 4-87 spots from a total of 131 IgE-binding protein spots. At least 12 Cry j 1 (27.5-75% of IgE-binding frequency) and 3 Cry j 2 (32.5-40%) isoforms were localized. In total, 31 spots were found to be more reactive than the highest IgE-reactive isoform of Cry j 2.
The proteomics approaches showed great interindividual variation of IgE-binding patterns to C. japonica proteins and contributed to the repertoire of numerous C. japonica allergens other than Cry j 1 and Cry j 2. Protein microsequencing demonstrated more complicated multiplicity in Cry j 1 than previously known and new isoforms in Cry j 2.
日本柳杉（Cryptomeria japonica）花粉是引发鼻炎和结膜炎的最常见过敏原来源之一。只有Cry j 1和Cry j 2被充分鉴定为该花粉的主要过敏原。
本研究的目的是完善日本柳杉花粉过敏原的种类，研究它们在IgE反应模式方面的变异性，并通过蛋白质组分析鉴定Cry j 1和Cry j 2的亚型。
在二维（2-D）聚丙烯酰胺凝胶电泳上分离的日本柳杉花粉中的蛋白质，用40名对日本柳杉花粉过敏的受试者血清中的IgE进行免疫检测。采用质谱指纹图谱法阐明主要过敏原的多样性。
用个体患者血清进行的二维免疫标记显示，在总共131个IgE结合蛋白斑点中，有4-87个斑点呈现出可区分的IgE结合模式。至少定位了12种Cry j 1亚型（IgE结合频率为27.5-75%）和3种Cry j 2亚型（32.5-40%）。总共发现31个斑点比Cry j 2的最高IgE反应性亚型反应性更强。
蛋白质组学方法显示，对日本柳杉蛋白质的IgE结合模式存在很大的个体差异，并有助于确定除Cry j 1和Cry j 2之外的众多日本柳杉过敏原种类。蛋白质微量测序表明，Cry j 1的多样性比以前已知的更为复杂，Cry j 2中存在新的亚型。
