Effect of fMLF on monocyte response to CCR5 ligands.
(A) Monocytes preincubated with fMLF (10−6 mol/L, 37°C) for the designated time intervals were tested for migration in response to MIP-1β (100 ng/mL). Cells preincubated with calphostin C (50 ng/mL, 2.5 hours, 37°C) followed by fMLF treatment were tested in parallel. Results are expressed as the net number of migrated monocytes in 3 high-powered fields (HPFs) after subtraction of spontaneous migration in response to medium (55 ± 7 cells) (P < .01). (B) Calcium mobilization of monocytes in response to fMLF and chemokines. Monocytes were preincubated with medium (Medium), calphostin C (50 ng/mL, 2.5 hours), followed by fMLF (10−6 mol/L, 37°C, 60 minutes) (C + fMLF), or fMLF (10−6 mol/L, 37°C, 60 minutes) (fMLF); the Ca++ mobilization in response to fMLF (10−6 mol/L) or the chemokines MIP-1α or RANTES (1 ng/mL) was then measured.
