Modulation of chemokine gene expression following T-cell activation.

Human PBLs were unstimulated or stimulated with PMA+Iono or soluble anti–(α-)CD3 mAb, soluble α-CD28 mAb plus PMA for 0, 2, 6, 12, 24, 48, and 72 hours, as indicated. Total RNA was prepared using Trizol, quantitated using OD₂₆₀, and analyzed by an RNase protection assay (RPA; see “Materials and methods”). (A) Results of 1 of 2 separate experiments, each using cells from different donors, are shown. MCP-1 was faintly detected but was not observed in subsequent experiments. (B) Chemokine mRNA levels normalized to levels of L32 mRNA were quantitated by PhosphorImager analysis using ImageQuant software.