High-precision gene expression analysis of immunosuppressive agents in human whole blood

✍ Scribed by Victor V. Tryon; John C. Cheronis; David B. Trollinger; Danute Bankaitis-Davis; Kathleen Storm; John S. Zuzack; Min Zhan; Dennis G. Macejak; Michael P. Bevilacqua

• Publisher: John Wiley and Sons
• Year: 2004
• Tongue: English
• Weight: 385 KB
• Volume: 63
• Category: Article
• ISSN: 0272-4391
• DOI: 10.1002/ddr.10348

✦ Synopsis

Abstract

Analysis of gene expression is emerging as a valuable tool in drug discovery. Using a gene expression system involving high‐precision quantitative PCR, two robust in vitro models of inflammation in human whole blood were examined. In the first model, lipopolysaccharide (LPS) addition to whole blood resulted in profound changes in the expression of multiple inflammation‐related genes, ranging from increases of 1.5‐fold to several 1,000‐fold. In the second model, stimulation by interleukin 2 plus toxic shock syndrome toxin 1 (IL2+TSST1) resulted in a distinct pattern of expression consistent with increased contributions from T‐lymphocyte activation. Both stimuli resulted in unique, but consistent patterns of gene expression across individual blood donors. In addition, experiments using the immunosuppressive drugs, dexamethasone and cyclosporin A, demonstrated that the distinct mechanisms of different immunosuppresants can be characterized at the molecular level in human whole blood using specific pathway markers. Thus, dexamethasone inhibited induction of gene expression at a number of loci in both models of the inflammatory response, while Cyclosporin A had minimal effects in the LPS model but had profound effects in the IL2+TSST1 model. These findings validate the use of high‐precision gene expression analysis (HPGEA) directly from human whole blood, coupled with well‐defined stimulation parameters, for characterizing, monitoring, and predicting the effects of novel therapeutic agents in pre‐clinical studies. Drug Dev. Res. 63:8–21, 2004. © 2004 Wiley‐Liss, Inc.