Effects of ADI on IEC numbers.

Abstract

<p>Triplicates of IEC numbers were monitored by MTT assay. In A, IECs (Caco2 and HCT-8 cells), were treated with medium only (medium, white bars), with WB trophozoites (WB, black bars) or with WB trophozoites overexpressing ADI (WB-ADI, grey bars) and measured after 4 d. In B, two silver stained SDS-PAGE gels are shown visualizing the purified ADI and OCT at the expected Mw of 65 kDA and 40 kDA respectively (arrows). In C and D, TC7 and HCT-8 cells were treated with 27.5 U/L ADI from <i>Giardia</i> and measured after 4 d (TC7 cells) and 40 h (HCT-8 cells). ADI induced effects were abolished by addition of arginine to 0.4 mM (ADI +Arg). Corresponding controls are given (buffer; ADIb, which is the boiled protein control; Arg that shows that arginine addition alone does not affect the cells). Cell numbers are expressed as relative to buffer ctrl, P-values are displayed. Note the reduction in cell number upon addition of ADI.</p