Fig. 4.
Type I (RH strain) T. gondii suppress ΗΒD2 gene expression in human intestinal epithelial cells. IEC were cultured in 24-well plates at a density of 3 × 105 cells/well and infected with the three T. gondii genotype strains at a MOI of 5. RNA was extracted from infected and noninfected cells at 3, 8, and 24 h postinfection. First-strand cDNA was synthesized using oligo(dT) primers and a RevertAid H Minus cDNA synthesis kit (Fermentas). Quantitative PCR was performed using primers specific for ΗΒD1 (A), ΗΒD2 (B), or ΗΒD3 (C) using SYBR green supermix (Bio-Rad). (D) Bradyzoites obtained from 76K-infected mice were used for infection in IEC, and HBD2 gene expression was measured at 3- and 8-h time points using the method described above. Results are expressed as relative gene expression levels. ***, P < 0.0001. Data are representative of two experiments, each performed in triplicate at each time point.