诱导小鼠淋巴细胞表达γ-干扰素的血吸虫成虫可溶性抗原组分  

Schistosome soluble adult worm antigen fractions inducing the spleen lymphocytes of mice to express interferon-gamma

作　　者：王薇[1] 何立[1] 吴栋才[1] 金烈[1] 宗红英[1] 张培喜[1] 

机构地区：[1]武汉大学基础医学院寄生虫学教研室,430071

出　　处：《中国地方病学杂志》2006年第5期487-489,共3页Chinese Jouranl of Endemiology

基　　金：国家自然科学基金(30471509)

摘　　要：目的 分离纯化日本血吸虫成虫可溶性抗原（soluble adult worm antigens，SAWA）各个组分，并分别鉴定其诱导小鼠脾淋巴细胞表达γ-干扰素（IFN-γ）的能力。方法 常规制备血吸虫SAWA，经十二烷基磺酸钠一聚丙烯酰胺凝胶电泳（SDS—PAGE）层析法分离、收集并透析纯化各抗原组分，SDS—PAGE检测分析其含量及相对分子质量（Mr）。常规制备感染42d小鼠脾淋巴细胞悬液，设立PBS阴性对照组及各组分抗原实验组，分别刺激体外培养的感染小鼠脾淋巴细胞，诱生IFN-γ，用双抗夹心酶联免疫吸附实验（ELISA）检测各组IFN-γ水平，筛选出能诱导IFN-γ高表达的抗原组分。结果 获得20种胁不同且含量较高的SAWA组分抗原，Mr（×10^3）分别为22，23、24、28、40、45、56、62、64、65、68、70、71、74、78、91、95、100、102、105，其电泳图谱均显示为清晰的单一带。各组分抗原均能不同程度地诱导IFN-γ表达，其中Mr（×10^3）为22、24、45、62、64、65的组分抗原诱导IFN-γ的表达量较高，尤其是62×103组分抗原诱导表达IFN-γ水平显著高于其他实验组（P＜0．01）。结论 电泳层析法能很好地将日本血吸虫成虫可溶性抗原分成多个组分：62×10^3组分抗原具有诱导小鼠脾淋巴细胞高表达IFN-γ的能力，推测它可能是一个很强的Th1型淋巴细胞活性诱导分子。Objective To fractionate the soluble adult worm antigens （SAWA） of Schistosomajaponicum and to screen the fractionated antigens inducing the spleen lymphocytes of infected mice to express interferon-gamma （IFN-γ） at high level. Methods SAWA of Schistosomajaponicum were prepared by routine method, and then were fractionated and collected by SDS-PAGE chromatography, and the relative molecular mass of these antigen fractions was determined by SDS-PAGE. The negative control group （PBS group） and a series of test groups （fractionated antigens） were designed to stimulate spleen lymphocytes of infected mice to express IFN-γ in vitro. The IFN-γ expression levels were assayed by ELISA. Then the fractionated antigens inducing the lymphocytes to express IFN-γ at high level were screened. Results Twenty fractionated antigens were obtained. SDS-PAGE showed that each fractionated antigens had a single protein band. Their relative molecular mass （Mr, × 10^3） was 22,23,24,28, 40,45,56,62,64,65,68,70,71,74,78,91,95,100,102 and 105, respectively. All fractionated antigens could induce the spleen lymphocytes of infected mice to express IFN-γ at different level. Fractionated antigens of （22,24,45,62,64,65） × 10^3 could induce IFN-γ at high level. The IFN-γ expression level induced by 62 × 10^3 antigen was the highest （P〈0.01 ）. Conclusions SAWA of Schistosorna japonicum has been fraetionated well by electrophoretic chromatography. The fractionated antigen of 62 × 10^3 could induce the lymphocytes of mice to express IFN-γ at high level. These findings suggest that 62 × 10^3 SAWA fractionated antigen of Schistosornajaponicum maybe a good inducer of Thl lymphocyte activity.

关 键 词：血吸虫 日本 抗原 电泳 聚丙烯酰胺凝胶 干扰素Ⅱ型 

分 类 号：R392[医药卫生—免疫学]