Molecular mechanisms of dissociative glucocorticoid activity.
BACKGROUND
Glucocorticoids mediate their effects on target cells via transactivation and transrepression of certain target genes. While conventional glucocorticoids do not distinguish between transactivation and transrepression, new glucocoticoids should be able to dissociate these effects, thus lowering the potential of unwanted side-effects of glucocorticoids in clinical use. In this study, we developed a new experimental system to test potentially selective glucocorticoids in normal lymphocytes.


METHODS
Following pretreatment with phytohaemagglutinin, normal lymphocytes were transfected, using electroporation, with pGL3 luciferase reporter vectors under the control: (1) of the human IL-2 promoter; and (2) of a glucocorticoid response element (GRE). Luciferase activity was measured in response to various steroid compounds, including the potentially dissociative glucocorticoid medroxyprogesterone acetate (MPA).


RESULTS
The IL-2 promoter was induced 267.2 +/- 27.5-fold (mean +/- SD) by phorbol ester and ionomycin. In these cells, hydrocortisone and dexamethasone caused a 22.9 +/- 3.6% and a 38.4 +/- 10% reduction in luciferase activity, respectively. Under GRE control, hydrocortisone stimulated luciferase activity 6.4 +/- 0.50-fold and dexamethasone 8.2 +/- 0.4-fold. MPA-induced transrepression was 73.3 +/- 7.2% for the IL-2 promoter, and transactivation was 2.4 +/- 0.4-fold with the GRE-driven construct. The natural progestin progesterone did not have significant effects on either construct.


CONCLUSIONS
This is the first system that allows efficient analysis of glucocorticoid-dependent transactivation and transrepression in normal human lymphocytes. Compared to conventional glucocorticoids, MPA can be referred to as a dissociative glucocorticoid, its transrepression/transactivation ratio being 6.6 (transrepression 1.91/transactivation 0.29), with dexamethasone being the standard (transrepression 1/transactivation 1). We conclude that MPA is a highly promising substance for the treatment of autoimmune/inflammatory diseases.
Bamberger
C M
CM
University Hospital Eppendorf, Department of Medicine, Hamburg, Germany.
Schulte
H M
HM
D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology; D007376:Interleukin-2 / Q000235:genetics; D017258:Medroxyprogesterone Acetate / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D015533:Transcriptional Activation / Q000187:drug effects*; D014162:Transfection
D005938:Glucocorticoids; D007166:Immunosuppressive Agents; D007376:Interleukin-2; D017258:Medroxyprogesterone Acetate