Adherence of Giardia lamblia trophozoites to Int-407 human intestinal cells.
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.
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. mcsousa@ci.uc.pt
M C
MC
Sousa
C A
CA
Gonçalves
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Bairos
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J
Poiares-Da-Silva
D000818:Animals; D000969:Antinematodal Agents / Q000494:pharmacology; D002448:Cell Adhesion / Q000502:physiology; D002614:Chelating Agents / Q000494:pharmacology; D003078:Colchicine / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D015638:Cytochalasin D / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D004492:Edetic Acid / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000469:parasitology*; D005404:Fixatives; D016829:Giardia lamblia / Q000254:growth & development* / Q000378:metabolism* / Q000648:ultrastructure; D005976:Glutaral; D006801:Humans; D007082:Ileum / Q000166:cytology*; D066298:In Vitro Techniques; D007583:Jejunum / Q000166:cytology*; D037102:Lectins / Q000378:metabolism; D008360:Mannosephosphates; D008463:Mebendazole / Q000494:pharmacology; D008854:Microscopy, Electron; D008855:Microscopy, Electron, Scanning; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology
D000969:Antinematodal Agents; D002614:Chelating Agents; D005404:Fixatives; D037102:Lectins; D008360:Mannosephosphates; D019384:Nucleic Acid Synthesis Inhibitors; D015638:Cytochalasin D; C027693:mannose-6-phosphate; D003571:Cytochalasin B; D008463:Mebendazole; D004492:Edetic Acid; D003078:Colchicine; D005976:Glutaral