In vitro model of attachment of Giardia intestinalis trophozoites to IEC-6 cells, an intestinal cell line

Abstract

Attachment of giardias to intestinal cells has been difficult to study because of a lack of a convenient in vitro model. We developed an assay for attachment of radiolabeled trophozoites to IEC-6 cells that can be done in microtiter trays. Attachment was confirmed by scanning and transmission electron microscopy. Trophozoites remained attached to the IEC-6 cells for 24 h with little evidence of damage to the IEC-6 cells. Preincubation of trophozoites with cytochalasins A, B, and D reduced attachment to approximately 20% of that of controls, whereas colchicine had no effect. Chelation of divalent cations with EDTA and EGTA [ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid] reduced attachment to 24 and 26% of control values, respectively, and incubation at 4 degrees C reduced attachment to 7% of the value for controls incubated at 37 degrees C. Glutaraldehyde fixation of trophozoites or IEC-6 cells resulted in significantly diminished attachment to the live substrate (17 and 40% of control values, respectively). Coincubation of IEC-6 cells and trophozoites on a rotary shaker resulted in detachment of 40% of trophozoites, but EDTA, EGTA, glutaraldehyde fixation of trophozoites, and low temperature diminished attachment markedly and significantly. Similar results were obtained in selected experiments with three strains of giardia.