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旋毛虫:一种由单克隆抗体鉴定出的76千道尔顿排泄/分泌幼虫抗原。

Trichinella spiralis: a 76-kDa excretory/secretory larval antigen identified by a monoclonal antibody.

作者信息

Kehayov I R, Kyurkchiev S D, Tankov C V, Komandarev S K

机构信息

Institute of Immunology, Bulgarian Academy of Sciences, Sofia.

出版信息

Exp Parasitol. 1991 Aug;73(2):150-60. doi: 10.1016/0014-4894(91)90018-r.

Abstract

Spleen cells from BALB/c mice were fused with myeloma cells following infection of the mice with Trichinella spiralis larvae and an ip booster injection with larval homogenate antigen. A monoclonal antibody (Mab), designated as TS 3G6 which did not react with sera or tissue extracts from noninfected mice, rats, and guinea pigs, was selected for further studies because of its high activity and specificity. When tested in ELISA TS 3G6 did not cross-react with Ascaris suum, A. lumbricoides, Toxocara canis, E. granulosus (larvae), Trichiuris suis, or T. ovis. Western blot analysis showed that Mab 3G6 recognized an antigen of 76 kDa located in the stichosome of the larvae as well as on the surface of the larval cuticle. Digestion of a larval extract with different enzymes suggests that the Mab TS 3G6 corresponding epitope is a polypeptide. The TS 3G6 antigen was detected in culture supernatants of Trichinella muscle larvae and in sera of experimentally infected animals using a sensitive ELISA assay. This secretory antigen also seemed to induce a specific immune response in the host since sera from infected animals could block the binding of Mab TS 3G6 to its target antigen when tested in a competitive ELISA.

摘要

用旋毛虫幼虫感染BALB/c小鼠,然后腹腔注射幼虫匀浆抗原加强免疫,之后将小鼠的脾细胞与骨髓瘤细胞进行融合。筛选出一种单克隆抗体(Mab),命名为TS 3G6,它不与未感染小鼠、大鼠和豚鼠的血清或组织提取物发生反应,因其具有高活性和特异性而被选作进一步研究。在ELISA检测中,TS 3G6不与猪蛔虫、人蛔虫、犬弓首蛔虫、细粒棘球绦虫(幼虫)、猪鞭虫或绵羊鞭虫发生交叉反应。蛋白质免疫印迹分析表明,单克隆抗体3G6识别位于幼虫杆状体以及幼虫角质层表面的一种76 kDa的抗原。用不同酶消化幼虫提取物表明,单克隆抗体TS 3G6对应的表位是一种多肽。使用灵敏的ELISA检测法在旋毛虫肌幼虫的培养上清液和实验感染动物的血清中检测到了TS 3G6抗原。这种分泌性抗原似乎也能在宿主体内诱导特异性免疫反应,因为在竞争性ELISA检测中,感染动物的血清能阻断单克隆抗体TS 3G6与其靶抗原的结合。

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