Table 1.
Correlation of parasite-induced activation of NF-κB in different cell types with resistance to infection
| Cell type | Cell line | p65 nuclear translocation (% positive) | Parasites/100 cells |
|---|---|---|---|
| Epithelial | Mv1Lu | 54.5 ± 0.4 | 22.2 ± 2.4 |
| Endothelial | SVEC4-10 | 51.9 ± 0.7 | 41.2 ± 5.1 |
| Fibroblast | Human primary | 18.0 ± 1.6 | 58.4 ± 4.1 |
| Muscle | Primary bovine aorta | 4.9 ± 0.5 | 136.2 ± 5.4 |
| Smooth | Primary human vascular | 4.5 ± 0.7 | 137.4 ± 8.9 |
| Skeletal | L6E9 | 3.5 ± 0.76 | 139.1 ± 4.3 |
| Cardiac | H9C2 | 3.1 ± 0.5 | 178.8 ± 2.1 |
Cells were incubated with trypomastigotes at 1 × 107/ml for 1 h, fixed, and stained with anti-p65 antibody as described in MATERIALS AND METHODS. Positively stained nuclei were counted for at least 200 cells. Nuclear localization was confirmed by DAPI staining. Results represent the mean of triplicate assays ± SEM. Background levels of nuclear p65 were <5% in all cases. Infection assays were carried out in 96-well plates at a host cell:parasite ratio of 1:5. After 1 h of incubation with parasites, the cells were washed, placed in fresh medium, and then incubated at 37°C for 2 d. After fixing and staining, the number of parasites per cell were counted in at least 300 cells. Results represent the mean of triplicate assays ± SEM. A similar pattern was observed when infection was measured in terms of percentage of cells infected. Consistent infection levels were obtained in multiple repeated experiments.