摘要
目的 分析草花粉变应原成分及变应原性、免疫原性。方法 制备草花粉变应原浸液 ,经十二烷硫酸钠 聚丙烯酰胺凝胶电脉 (SDS PAGE)分离 ,部分考马斯亮蓝染色 ,其余电转印至硝酸纤维素薄膜上 ,封闭后将血清 1∶10稀释与膜条孵育 ,洗涤后分别与辣根过氧化物酶标兔抗人IgE单克隆抗体 (IgE HRP)、辣根过氧化物酶标兔抗人IgG单克隆抗体 (IgG HRP)反应 ,加底物 4 氯 1 萘酚显色。结果 草花粉蛋白电泳清晰地显示出 8条区带 ,蛋白分子量依次为 10 0 .5、76 .2、6 5 .0、5 5 .0、4 2 .0、35 .1、2 8.8和 2 3.2～ 11.8kD ,免疫印迹可见 5条sIgE反应带 ,蛋白分子量依次为 10 0 .5、76 .2、35 .1、2 8.8和 8.0kD ,结合病人血清百分比分别是 30 .8%、7.7%、3.8%、11.5 %和 84 .6 % ;与血清sIgG结合的反应带有 5条 ,蛋白分子量为 10 0 .5、76 .2、4 9.8、2 8.8、8.0kD ,结合病人血清百分比分别是96 .2 %、7.7%、7.7%、3.8%和 7.7%。结论 草花粉变应原含 8种主要蛋白成分 ;此外至少还有 2种含量较低的蛋白成分 ;草花粉变应原中分子量 10 0 .5、76 .2、35 .1、2 8.8、8.0kD的蛋白具有免疫原活性 。
Humulus pollen is considered to be a very important allergen in China. In order to analyse protein components in humulus pollen and evaluate their allergenicity and immunogenicity, crude pollen extracts was prepared and was separated by SDS PAGE. The gel was divided into 2 shares, one for visualizing protein bands stained with Coomassie brilliant blue R250, the other part was transferred onto nitrocellulose membrane. After blocked with milk powder solution, membranes were incubated with the serum sample diluted 1∶10. The strips were then washed, followed by additional 1 hour incubation with HRP conjugated rabbit anti human IgE?IgG, respectively. 4 chloride 1 naphtol was added as substrate. The SDS-PAGE gel stained with Coomassie brilliant blue R250 illustrated 8 bands clearly, their mol. wts were 100.5, 76.2, 65.0, 55.0, 42.0, 35.1, 28.8 and 23.2～11.8kD. Immunobloting identified 5 protein bands recognized by sIgE with mol. wts 100.5, 76.2, 35.1, 28.8 and 8.0kD, which combined with 30.8%, 7.7%, 3.8%, 11.5% and 84.6% of asthmatic patients serum, respectively. 5 protein bands with mol. wts 100.5, 76.2, 49.8, 28.8 and 8.0kD showed sIgG binding capacity, which reacted with 96.2%, 7.7%, 7.7%, 3.8% and 7.7% of asthmatic patients serum. Our results indicate that humulus pollen includes 8 major protein components and at least 2 minor protein components. Components with allergenic activity are protein with mol. wts 100.5, 76.2, 35.1, 28.8 and 8.0kD, which can cause immediate asthma reaction. In addition protein with mol. wts 100.5, 76.2, 49.8, 28.8, and 8.0kD have immunogenic activity,and they can induce production of protective antibodies during specific immunotherapy of asthma. However, they may also play a direct role in asthma.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2001年第S2期28-30,共3页
Chinese Journal of Microbiology and Immunology
关键词
哮喘
葎草花粉
变应原
变应原性
免疫原性
Asthma
Humulus pollen
Allergen
Allergenicity
Immunogenicity
