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制备单克隆抗体以鉴定卫氏并殖吸虫抗原。

Production of monoclonal antibody to characterize the antigen of Paragonimus westermani.

作者信息

Sugiyama H, Hinoue H, Katahira J, Horiuchi T, Tomimura T, Kamata Y, Kozaki S

机构信息

Laboratory of Veterinary Pathology, College of Agriculture, University of Osaka Prefecture, Japan.

出版信息

Parasitol Res. 1988;75(2):144-7. doi: 10.1007/BF00932714.

Abstract

We produced monoclonal antibodies against the adult antigen of Paragonimus westermani to investigate the expression of the stage-specific antigen of adult flukes. Two hybridoma cell lines, A1-2 and A4-1, were established. A1-2 reacted specifically only with the adult antigen in an enzyme-linked immunosorbent assay (ELISA), whereas A4-1 reacted with both adult and larval antigens. By immunoblotting analysis, A1-2 was found to react with four bands with molecular weights of 35,000, 17,000, 15,500, and 12,500 in the adult antigen but with none in the larval antigen. A4-1 was found to bind to the band of 27,000 daltons in the adult antigen as well as those of 28,000 and 26,000 daltons in the larval antigen. By immunohistochemical methods, a positive reaction was observed only in the parenchymal tissues of the adult flukes with A1-2. A4-1 reacted with the substance that located on the gut epithelium and in the luminal contents of both the adult and larval flukes. These results indicate that the adult P. westermani possesses an antigen or antigenic determinant specific to the adult stage, as well as that common to both the larval and adult stages.

摘要

我们制备了针对卫氏并殖吸虫成虫抗原的单克隆抗体,以研究成虫期特异性抗原的表达。建立了两个杂交瘤细胞系,A1 - 2和A4 - 1。在酶联免疫吸附测定(ELISA)中,A1 - 2仅与成虫抗原发生特异性反应,而A4 - 1则与成虫和幼虫抗原均发生反应。通过免疫印迹分析发现,A1 - 2与成虫抗原中分子量为35,000、17,000、15,500和12,500的四条带发生反应,但与幼虫抗原无反应。A4 - 1被发现与成虫抗原中27,000道尔顿的条带以及幼虫抗原中28,000和26,000道尔顿的条带结合。通过免疫组织化学方法,仅在成虫吸虫的实质组织中观察到A1 - 2的阳性反应。A4 - 1与位于成虫和幼虫吸虫的肠上皮及管腔内容物中的物质发生反应。这些结果表明,卫氏并殖吸虫成虫具有成虫期特异性的抗原或抗原决定簇,以及幼虫和成虫期共有的抗原。

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