| 藜草花粉变应原免疫活性分析
| | 中药学论文 作者:张洁,孙秀珍,陈曦,冯向莉,刘艳琴 【关键词】 哮喘;藜草;花粉;变应原;变应原性;免疫原性 Immunoactivity analysis of album pollen allergen 【Abstract】 AIM: To analyse protein components in album pollen and evaluate their allergenicity and immunogenicity. METHODS: The components of protein allergen were collected from the crude extract of chenopodium album pollen with Sephacryl S200 HR gel filtration for removing the impurities. Molecular weights of the components were tested through SDSPAGE. Then the reaction rates of IgE and IgG in asthmatic patients serum to chenopodium album pollen were counted by immunoblotting. RESULTS: Twelve bands of protein were obtained through SDSPAGE. The molecular weight of each band was 92, 63.1, 61, 52, 45, 43, 39.7, 38.9, 34, 31.6, 28.4 and 18.5~12 ku. Immunobloting identified 4 protein bands recognized by sIgE, molecular weights being 92, 34, 31.6 and 18.5~12 ku, which reacted with 70%, 50%, 80% and 90% of asthmatic patient serum. Eight protein bands with molecular weights being 92, 61, 52, 45, 43, 39.7, 31.6 and 18.5~12 ku showed sIgG binding capacity, which reacted with 80%, 40%, 10%, 20%, 80%, 10%, 10% and 100% of asthmatic patient serum. CONCLUSION: Album pollen comprises 12 major protein components. Components with strong allergenicity and immunogenicity are the proteins with molecular weight of 18.5~12 and 92 ku. Proteins with molecular weight of 34 and 31.6 ku have strong allergenicity, and weak immunogenicity or not. Proteins with molecular weight of 61,43,45,52 and 39.7 ku possess certain immunogenicity, but no allergenicity. 【Keywords】 asthma; chenopodium album; pollen; allergens; allergenicity; immunogenicity 【摘要】 目的中药学论文: 分析藜草花粉变应原成分及其变应原性、免疫原性. 方法: 用Sephacryl S200HR葡聚糖凝胶层析法分离、纯化藜草花粉变应原,通过十二烷硫酸钠聚丙烯酰胺凝胶电脉法(SDSPAGE)测定变应原各组分的中药学论文分子质量;应用免疫印迹法检测各组分蛋白质与藜草花粉过敏性哮喘患者血清中sIgG和sIgE的反应率. 结果: 藜草花粉经电泳后得到12条蛋白质区带,分子质量依次为92, 63.1, 61, 52, 45, 43, 39.7, 38.9, 34, 31.6, 28.4和18.5~12 ku;免疫印迹可见4条sIgE反应带,蛋白分子质量依次为92, 34, 31.6和18.5~12 ku;与患者血清反应率分别是70%, 50%, 80%和90%;与血清sIgG结合的反应带有8条,蛋白分子质量为92, 61, 52, 45, 43, 39.7, 31.6和18.5~12 ku;与患者血清反应率分别是80%, 40%, 10%, 20%, 80%, 10%, 10%和100%. 结论: 藜草花粉变应原主要含12种蛋白质成分,其中分子质量为18.5~12 ku和92 ku的蛋白质变应原活性及免疫原活性均很强,是主要变应原成分;分子质量在34和31.6 ku的蛋白质有较强的变应原性而无或仅有较弱的免疫原性;而分子质量为43, 61, 45, 52和39.7 ku的蛋白质具有一定的免疫原性而无变应原性. 【关键词】 哮喘;藜草;花粉;变应原;变应原性;免疫原性 0引言 从气传致敏花粉种类调查中发现,藜草是我国许多地方夏秋季节大气中最常见的气传致敏花粉之一[1]. 我们采用SDSPAGE分析藜草花粉变应原的蛋白成分,并用免疫印迹的方法,通过观察藜草花粉过敏性哮喘患者的血清结合模式,分析藜草花粉各蛋白成分的变应原性及免疫原性,为藜草花粉变应原的纯化、重组打下基础. 1材料和方法
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