A method for visualizing surface-exposed and internal PfEMP1 adhesion antigens in Plasmodium falciparum infected erythrocytes.
A method for visualizing surface-exposed and internal PfEMP1 adhesion antigens in Plasmodium falciparum infected erythrocytes.
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DOI：
10.1186/1475-2875-7-101
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发表时间：
2008-06-05
期刊：
影响因子：
3
通讯作者：
Arnot DE
中科院分区：
文献类型：
--
作者：
Bengtsson D;Sowa KM;Salanti A;Jensen AT;Joergensen L;Turner L;Theander TG;Arnot DE
关于寄生虫抗原插入宿主红细胞膜以及恶性疟原虫黏附受体在红细胞膜上的结构和分布，人们知之甚少。利用激光扫描共聚焦显微镜(LSCM)和一种新的标记和固定方法，获得了红细胞膜表面PfEMP1和内部抗原的高分辨率免疫荧光图像，从而可以分析PfEMP1在无性发育过程中在红细胞膜上的积聚。一种新的染色技术已经被开发出来，它允许在其核物质被异常良好地分辨的寄生虫中区分红细胞表面PfEMP1和细胞内PfEMP1。对活体寄生红细胞进行一次抗体荧光检测。然后用多聚甲醛固定表面标记的细胞，并用非离子洗涤剂渗透，以允许抗体进入内部寄生虫抗原。通过用不同荧光染料标记的抗体和共聚焦显微镜来区分表面和内部抗原，表面暴露的PfEMP1首先在细胞内寄生虫发育的滋养体阶段被抗体检测到，尽管改进的检测方法表明不同的实验室分离株在表面暴露的PfEMP1的积累动力学上存在差异。为了激光扫描共聚焦光学显微镜和分析疟疾黏附抗原的发育表达，建立了一种灵敏的标记PfEMP1表面和内部多达三种不同荧光的方法。
The insertion of parasite antigens into the host erythrocyte membrane and the structure and distribution of Plasmodium falciparum adhesion receptors on that membrane are poorly understood. Laser scanning confocal microscopy (LSCM) and a novel labelling and fixation method have been used to obtain high resolution immuno-fluorescent images of erythrocyte surface PfEMP1 and internal antigens which allow analysis of the accumulation of PfEMP1 on the erythrocyte membrane during asexual development. A novel staining technique has been developed which permits distinction between erythrocyte surface PfEMP1 and intracellular PfEMP1, in parasites whose nuclear material is exceptionally well resolved. Primary antibody detection by fluorescence is carried out on the live parasitized erythrocyte. The surface labelled cells are then fixed using paraformaldehyde and permeabilized with a non-ionic detergent to permit access of antibodies to internal parasite antigens. Differentiation between surface and internal antigens is achieved using antibodies labelled with different fluorochromes and confocal microscopy Surface exposed PfEMP1 is first detectable by antibodies at the trophozoite stage of intracellular parasite development although the improved detection method indicates that there are differences between different laboratory isolates in the kinetics of accumulation of surface-exposed PfEMP1. A sensitive method for labelling surface and internal PfEMP1 with up to three different fluorochromes has been developed for laser scanning confocal optical microscopy and the analysis of the developmental expression of malaria adhesion antigens.
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DOI：
10.1097/00022744-200112000-00010
发表时间：
2001-12-01
影响因子：
1.6
作者：
Hoetelmans, RWM;Prins, FA;van Dierendonck, JH
通讯作者：
van Dierendonck, JH
影响因子：
20.3
作者：
Knuepfer, E;Rug, M;Cowman, AF
通讯作者：
Cowman, AF
影响因子：
20.3
作者：
Hassoun, H;Hanada, T;Chishti, AH
通讯作者：
Chishti, AH
影响因子：
3.6
作者：
Salanti, A;Staalsoe, T;Theander, TG
通讯作者：
Theander, TG
影响因子：
2
作者：
WEIBULL, C;CHRISTIANSSON, A;CARLEMALM, E
通讯作者：
CARLEMALM, E