Sousa M C, Gonçalves C A, Bairos V A, Poiares-Da-Silva J
Laboratory of Microbiology and Parasitology and Center of Pharmaceutical Studies, Faculty of Pharmacy, University of Coimbra, Couraça dos Apóstolos, n.51, r/c, 3030 Coimbra, Portugal.
Clin Diagn Lab Immunol. 2001 Mar;8(2):258-65. doi: 10.1128/CDLI.8.2.258-265.2001.
Attachment of Giardia lamblia trophozoites to enterocytes is essential for colonization of the small intestine and is considered a prerequisite for parasite-induced enterocyte dysfunction and clinical disease. In this work, coincubation of Giardia with Int-407 cells, was used as an in vitro model to study the role of cytoskeleton and surface lectins involved in the attachment of the parasite. This interaction was also studied by scanning and transmission electron microscopy. Adherence was dependent on temperature and was maximal at 37 degrees C. It was reduced by 2.5 mM colchicine (57%), mebendazole (10 microg/ml) (59%), 100 mM glucose (26%), 100 mM mannose (22%), 40 mM mannose-6-phosphate (18%), and concanavalin A (100 microg/ml) (21%). No significant modification was observed when Giardia was pretreated with cytochalasins B and D and with EDTA. Giardia attachment was also diminished by preincubating Int-407 cells with cytochalasin B and D (5 microg/ml) (16%) and by glutaraldehyde fixation of intestinal cells and of G. lamblia trophozoites (72 and 100%, respectively). Ultrastructural studies showed that Giardia attaches to the Int-407 monolayer predominantly by its ventral surface. Int-407 cells contact trophozoites with elongated microvilli, and both trophozoite imprints and interactions of Giardia flagella with intestinal cells were also observed. Transmission electron microscopy showed that Giardia lateral crest and ventrolateral flange were important structures in the adherence process. Our results suggest a combination of mechanical and hydrodynamic forces in trophozoite attachment; surface lectins also seem to mediate binding and may be involved in specific recognition of host cells.
蓝氏贾第鞭毛虫滋养体与肠上皮细胞的附着对于小肠定植至关重要,被认为是寄生虫诱导肠上皮细胞功能障碍和临床疾病的先决条件。在本研究中,将蓝氏贾第鞭毛虫与Int-407细胞共同孵育,用作体外模型来研究参与寄生虫附着的细胞骨架和表面凝集素的作用。这种相互作用也通过扫描电子显微镜和透射电子显微镜进行了研究。黏附依赖于温度,在37℃时达到最大值。2.5 mM秋水仙碱(57%)、甲苯咪唑(10 μg/ml)(59%)、100 mM葡萄糖(26%)、100 mM甘露糖(22%)、40 mM甘露糖-6-磷酸(18%)和伴刀豆球蛋白A(100 μg/ml)(21%)可使其降低。用细胞松弛素B和D以及EDTA预处理蓝氏贾第鞭毛虫时,未观察到明显变化。用细胞松弛素B和D(5 μg/ml)预孵育Int-407细胞(16%)以及用戊二醛固定肠道细胞和蓝氏贾第鞭毛虫滋养体(分别为72%和100%)也会减少蓝氏贾第鞭毛虫的附着。超微结构研究表明,蓝氏贾第鞭毛虫主要通过其腹面附着于Int-407单层细胞。Int-407细胞通过细长的微绒毛与滋养体接触,还观察到滋养体印记以及蓝氏贾第鞭毛虫鞭毛与肠道细胞的相互作用。透射电子显微镜显示,蓝氏贾第鞭毛虫的侧嵴和腹侧凸缘是黏附过程中的重要结构。我们的结果表明,滋养体附着过程中存在机械力和流体动力的结合;表面凝集素似乎也介导结合,并且可能参与宿主细胞的特异性识别。