体外培养的刚地弓形虫和犬新孢子虫囊肿的自体荧光。

Autofluorescence of Toxoplasma gondii and Neospora caninum cysts in vitro.

作者信息

Lei Ying, Davey Mary, Ellis John

机构信息

Department of Cell and Molecular Biology, University of Technology, Sydney, Gore Hill, New South Wales 2065, Australia.

出版信息

J Parasitol. 2005 Feb;91(1):17-23. doi: 10.1645/GE-355R.

DOI:10.1645/GE-355R

PMID:15856866

Abstract

Autofluorescence of Toxoplasma gondii and Neospora caninum was studied by fluorescence microscopy during their differentiation from tachyzoites to bradyzoites in vitro using Vero as host cells. Stage conversion into bradyzoites and cysts was confirmed by immunofluorescent microscopy and Western blot analysis using SAG1- and BAG1-specific antibody, respectively. From day 4 postinfection (PI), pale blue autofluorescence of the bradyzoites and tissue cysts was observed with UV light at 330-385 nm, which coincided with the onset of cyst development. This autofluorescence under UV light of bradyzoites and tissue cysts increased in intensity from days 8 to 10 PI. In contrast to the autofluorescence shown by bradyzoites and cysts, tachyzoites and parasitophorous vacuoles containing tachyzoites never autofluoresced at any time examined. Autofluorescence of the cystic stages was of sufficient intensity and duration to allow the detection of cysts and bradyzoites of T. gondii and N. caninum. In this study, we describe for the first time the autofluorescence properties of in vitro-induced bradyzoites and cysts of T. gondii and N. caninum.

摘要

利用Vero细胞作为宿主细胞，通过荧光显微镜研究了体外培养的刚地弓形虫和犬新孢子虫从速殖子分化为缓殖子过程中的自发荧光。分别使用SAG1和BAG1特异性抗体，通过免疫荧光显微镜和蛋白质印迹分析确认了向缓殖子和包囊的阶段转化。感染后第4天（PI）起，在330 - 385 nm紫外光下观察到缓殖子和组织包囊发出淡蓝色自发荧光，这与包囊发育的开始时间一致。缓殖子和组织包囊在紫外光下的自发荧光强度在感染后第8天至第10天增加。与缓殖子和包囊显示的自发荧光不同，速殖子以及含有速殖子的寄生泡在任何检测时间都未出现自发荧光。包囊阶段的自发荧光强度和持续时间足以检测到刚地弓形虫和犬新孢子虫的包囊和缓殖子。在本研究中，我们首次描述了体外诱导的刚地弓形虫和犬新孢子虫缓殖子和包囊的自发荧光特性。