Figure 1

The stimulatory effect of recombinant HSP70 on primary human macrophages is reduced by inhibition of TLR4, or by Polymyxin B treatment of the HSP70. (A) Primary human macrophages were left untreated (NT), or treated with 100 μg/mL of HTA125 or an isotype control antibody for 30 min. Macrophages treated with antibodies were then stimulated for 4 hr with rhHSP70 at a final concentration of 10 μg/mL. Cell culture supernatants were collected and analyzed for TNF-α. **p < 0.005. (B) 5 μg rhHSP70 was pretreated with buffer alone or 10 μg Polymyxin B (at 100 μg/mL) for 30 min on ice, and then used to stimulate primary human macrophages for 4 hr at 10 μg/mL rhHSP70 final concentration in each culture. Cell culture supernatants were collected and analyzed for TNF-α. *p < 0.05, ***p < 0.0005.