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单克隆抗体对蓝氏贾第鞭毛虫囊肿特异性抗原的识别。

Recognition of Giardia lamblia cyst-specific antigens by monoclonal antibodies.

作者信息

Campbell J D, Faubert G M

机构信息

Institute of Parasitology, McGill University, Ste-Anne-de-Bellevue, Québec, Canada.

出版信息

Parasite Immunol. 1994 Apr;16(4):211-9. doi: 10.1111/j.1365-3024.1994.tb00342.x.

Abstract

Immunization of BALB/c mice with a sonicated extract of in vitro-generated Giardia lamblia cysts produced six cyst-specific monoclonal antibodies (MoAbs). Two MoAbs (8C5.C11 and 5A4.G6), which recognize proteinaceous cyst antigens, were selected for further study. In indirect immunofluorescence (IFA), MoAb 8C5.C11 reacted with encystation-specific vesicles in trophozoites beginning 3 h after the induction of encystation in vitro. This MoAb also recognized cysts which began to appear at 12 h. In contrast, MoAb 5A4.G6 stained only cyst walls. In Western blots, both MoAbs also reacted with cyst antigens, but not trophozoite antigens. MoAb 8C5.C11 first recognized cyst antigen from 3 h encysting cultures, reacting with 26, 28, 42 and 46 kD bands. MoAb 5A4.G6 reacted with a 38 kD band, beginning with 12 h encysting cultures. When added to G. lamblia encysting cultures before the appearance of cysts (0 to 9 h) and in the presence of a source of complement, MoAb 8C5.C11 caused a significant reduction in the numbers of water-resistant cysts produced in vitro compared to the control. MoAb 5A4.G6 did not affect in vitro encystation. These findings confirm the heterogeneity of cyst antigens, and also indicate that the process of encystation in vitro can be interrupted by antibodies and complement.

摘要

用体外培养的蓝氏贾第鞭毛虫囊肿的超声提取物免疫BALB/c小鼠,产生了六种囊肿特异性单克隆抗体(MoAbs)。选择了两种识别蛋白质性囊肿抗原的单克隆抗体(8C5.C11和5A4.G6)进行进一步研究。在间接免疫荧光法(IFA)中,单克隆抗体8C5.C11在体外诱导包囊化3小时后开始与滋养体中的包囊化特异性囊泡发生反应。该单克隆抗体也识别在12小时开始出现的囊肿。相比之下,单克隆抗体5A4.G6仅对囊肿壁进行染色。在蛋白质印迹法中,两种单克隆抗体也都与囊肿抗原发生反应,但不与滋养体抗原发生反应。单克隆抗体8C5.C11首先从培养3小时的包囊化培养物中识别出囊肿抗原,与26、28、42和46 kD条带发生反应。单克隆抗体5A4.G6从培养12小时的包囊化培养物开始与一条38 kD的条带发生反应。当在囊肿出现之前(0至9小时)且存在补体来源的情况下添加到蓝氏贾第鞭毛虫包囊化培养物中时,与对照组相比,单克隆抗体8C5.C11导致体外产生的耐水囊肿数量显著减少。单克隆抗体5A4.G6不影响体外包囊化。这些发现证实了囊肿抗原的异质性,也表明体外包囊化过程可被抗体和补体阻断。

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