Ex vivo stimulation of whole blood as a means to determine glucocorticoid sensitivity

J Inflamm Res. 2012:5:89-97. doi: 10.2147/JIR.S33569. Epub 2012 Aug 22.

Abstract

Purpose: Glucocorticoids are commonly prescribed to treat a number of diseases including the majority of inflammatory diseases. Despite considerable interpersonal variability in response to glucocorticoids, an insensitivity rate of about 30%, and the risk of adverse side effects of glucocorticoid therapy, currently no assay is performed to determine sensitivity.

Patients and methods: Here we propose a whole blood ex vivo stimulation assay to interrogate known glucocorticoid receptor (GR) up- and downregulated genes to indicate glucocorticoid sensitivity. We have chosen to employ real-time PCR in order to provide a relatively fast and inexpensive assay.

Results: We show that the GR-regulated genes, GILZ and FKBP51, are upregulated in whole blood by treatment with dexamethasone and that LPS-induction of cytokines (IL-6 and TNFα) are repressed by dexamethasone in a dose responsive manner. There is considerable interpersonal variability in the maximum induction of these genes but little variation in the EC(50) and IC(50) concentrations. The regulation of the GR-induced genes differs throughout the day whereas the suppression of LPS-induced cytokines is not as sensitive to time of day.

Conclusion: In all, this assay would provide a method to determine glucocorticoid receptor responsiveness in whole blood.

Keywords: FKBP51; GILZ; cytokines; gene regulation; glucocorticoid responsiveness; nuclear receptor.