FIGURE 8.

Interaction of Ecolihsp60 either with TLR2 or TLR4 results in induction of TNF-α. A, PMA-differentiated THP-1 macrophages were pretreated with 10 μg/ml of anti-TLR2 mAb or anti-TLR4 mAb or isotype-matched control antibody for 1 h, followed by incubation with Ecolihsp60-FITC (10 μg/ml) at 37 °C for 15 min. Cells were fixed and washed, and endocytosis of the protein was assessed by confocal laser scanning microscopy. B, PMA-differentiated THP-1 macrophages were pretreated with 10 μg/ml of anti-TLR2 mAb or anti-TLR4 mAb or isotype-matched control antibody for 1 h and then incubated with 3 μg/ml Ecolihsp60. TNF-α levels were quantified after 48 h in different culture supernatants by EIA. Data are representative of mean ± S.D. of three different experiments.