Roles of chemokines in attracting T cells and neutrophils. We incubated the neutralizing antibody to MIP-1α, TARC, or MIP-2 with the conditioned supernatant for 30 minutes. The supernatant was 10-fold diluted and used to chemoattract the mixed cells containing T cells stained with CD3-PE and neutrophils stained with Gr1-FITC in 3.0-μm pore sized transwell insert. After 2 hours of incubation at 37°C, the total cells (top), including Gr1-FITC neutrophils (left bottom) and CD3-PE T cells (right bottom) that had migrated into wells, were harvested and counted by FACSCalibur flow cytometer and analyzed by CellQuest software.