Characterization of excretory-secretory products from protoscoleces of Echinococcus granulosus and evaluation of their potential for immunodiagnosis of human cystic echinococcosis
Characterization of excretory-secretory products from protoscoleces of Echinococcus granulosus and evaluation of their potential for immunodiagnosis of human cystic echinococcosis
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DOI：
10.1017/s0031182004005670
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发表时间：
2004-09-01
期刊：
影响因子：
2.4
通讯作者：
Guisantes, JA
中科院分区：
文献类型：
--
作者：
Carmena, D;Martínez, J;Guisantes, JA
本研究首次描述了细粒棘球绦虫原头节的排泄-分泌抗原（ES-Ag）的特征，并评价了其在人囊性棘球绦虫病免疫诊断中的作用。ES-Ag是在原头节体外维持50 h后获得的。该制剂含有20多种主要蛋白质成分，可通过一维SDS-PAGE识别，表观质量在9 ~ 300 kDa之间。肝原头节的培养比肺原头节的培养产生更多种类的排泄-分泌蛋白成分。分泌蛋白的酶活性测定显示存在磷酸酶、脂肪酶和葡萄糖苷酶，但没有蛋白酶。这些发现与从原头节和包虫囊液产品的体细胞提取物中获得的结果进行了比较。免疫化学特性通过免疫印迹法对囊性包虫病（n = 15）、非包虫病寄生虫（n = 19）、各种肝脏疾病（n = 24）、肺肿瘤（n = 16）和健康供者（n = 18）感染的血清进行检测。89、74、47/50、32和20 kDa抗原对人包虫病的免疫诊断具有特异性。89和74 kDa成分对应于颗粒棘球绦虫中尚未描述的抗原，而41-43 kDa和91-95 kDa的蛋白质被研究的大多数非棘球绦虫血清识别。
This study describes, for the first time, the characterization of excretory-secretory antigens (ES-Ag) from Echinococcus granulosus protoscoleces, evaluating their usefulness in the immunodiagnosis of human cystic echinococcosis. ES-Ag were obtained from the first 50 h maintenance of protoscoleces in vitro. This preparation contained over 20 major protein components which could be distinguished by 1-dimensional SDS-PAGE with apparent masses between 9 and 300 kDa. The culture of of protoscoleces from liver produced a greater variety of excretory-secretory protein components than those from lung. Determination of enzymatic activities of secreted proteins revealed the presence of phosphatases, lipases and glucosidases, but no proteases. These findings were compared to those obtained from somatic extracts of protoscoleces and hydatid cyst fluid products. Immunochemical characterization was performed by immunoblotting with sera from individuals infected by cystic echinococcosis (n = 15), non-hydatidic parasitoses (n = 19), various liver diseases (it = 24), lung neoplasia (n = 16), and healthy donors (n = 18). Antigens with apparent masses of 89, 74, 47/50, 32, and 20 kDa showed specificity for immunodiagnosis of human hydatidosis. The 89 and 74 kDa components corresponded to antigens not yet described in E. granulosus, whereas proteins of 41-43 kDa and 91-95 kDa were recognized by the majority of the nonhydatid sera studied.