短穗鱼尾葵及长穗鱼尾葵花粉过敏原的分析、鉴定与纯化  

Characterization of the Antigenic Properties of Pollen Allergens in Carvota mitis and Caryota ochlandra

作　　者：刘晓宇[1] 肖淑英[1] 刘志刚[1] 姚敏[2] 孟光[2] 

机构地区：[1]深圳大学过敏反应与免疫学研究所,深圳518060 [2]海南省海口市人民医院耳鼻咽喉科,海口570208

出　　处：《热带医学杂志》2007年第10期952-955,977,共5页Journal of Tropical Medicine

基　　金：国家自然科学基金(No.30070702);广东省科技重点专项(No.2003A3080502);深圳市科技计划(No.200326);深圳大学校基金(No.200528)。

摘　　要：目的对长穗鱼尾葵和短穗鱼尾葵花粉变应原蛋白进行分析、鉴定与纯化。方法提取这两种花粉的粗提液,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离粗提液蛋白质组分并测定其分子量。收集过敏病人血清,采用免疫印迹(Western-blotting)法鉴定其变应原成分,通过离子交换层析对花粉变应原进行初步纯化和免疫印迹鉴定。结果长穗鱼尾葵花粉有30余条蛋白带,其中主要条带有18条,12000和14000 Mr为长穗鱼尾葵花粉特异性变应原;短穗鱼尾葵花粉有30余条蛋白带,其中主要条带有9条,26000、12000和14000 Mr为短穗鱼尾葵花粉特异性变应原,其中14000 Mr为主要变应原;通过离子交换层析方法纯化出长穗鱼尾葵花粉相对分子质量为12000和14000的变应原主要分布在Ⅱ峰中,短穗鱼尾葵花粉相对分子质量为14000的主要变应原分布在Ⅴ峰和Ⅵ峰中。结论对长穗和短穗鱼尾葵花粉变应原进行了初步的分离、鉴定和纯化,为临床鱼尾葵花粉变态反应疾病的诊断和治疗奠定了基础。Objective To characterize and purify the pollen allergens isolated from Carvota mitis and Caryota ochlandra. Method Pollen extracts from Carvota mitis and Caryota ochlandra were prepared and analyzed by SDSPAGE and proteins bands were visualized by staining the gel with Coomassis blue. Antigenic properties of the separated proteins were analyzed by immuno-blotting. Ion-exchange chromatography with DE-52 was used for the purification of the antigens. The purified products were characterized by Western-blotting. Result For the pollen extract of Caryota ochlandra, 30 protein bands were detected in SDS - PAGE. Two of these protein bands, with a molecular weight of 12 000 Mr and 14 000 Mr, showed immunoreactivity with IgE in the sera from patients with allergy to pollen of Caryota ochlandra. For the pollen extract of Carvota mitis, 30 protein bands were detected in SDS-PAGE, and three of these protein bands, with a molecular weight of 12 000, 14 000, and 26 000 Mr, showed immunoreactivity with IgE in the sera from patients with pollen allergy of Carvota mitis. The 14 000 Mr protein was the major allergen. Result from the ion-exchange chromatography indicated that the two pollen allergens （12 000, 14 000 Mr） from Caryota ochlandra were eluted in the second peak. The major pollen allergen of Carvota mitis was eluted in the forth and fifth peak. Conclusion Pollen allergens from Carvota mitis and Caryota ochlandra were purified and characterized. The result will provide a basis for the diagnosis and treatment of allergy to the pollen allergens from Carvota mitis and Caryota ochlandra.

关 键 词：短穗鱼尾葵花粉 长穗鱼尾葵 变应原 离子交换层析 

分 类 号：R392.8[医药卫生—免疫学]