title: Limited Applicability of GW9662 to Elucidate PPARγ-Mediated Fatty Acid Effects in Primary Human T-Helper Cells.
abstract: Synthetic antagonists of the nuclear receptor PPARγ such as GW9662 are widely used to elucidate receptor-mediated ligand effects. In addition and complementary to recent work, we examined whether GW9662 is suitable to serve for mechanistic investigation in T-helper cells. Human peripheral blood mononuclear cells (PBMC) were preincubated with increasing concentrations of GW9662 (0, 0.4, 2, and 10 μmol/L) 30 min before adding the c9,t11-isomer of conjugated linoleic acid (c9,t11-CLA) as representative of PPARγ-activating fatty acids with immunomodulatory properties. Corresponding cultures were incubated with GW9662 in the absence of the fatty acid. After 19 h, cells were mitogen stimulated for further 5 h. Subsequently, intracellular IL-2 was measured in CD3(+)CD4(+) lymphocytes by means of flow cytometry. 100 μmol/L c9,t11-CLA reduced the number of T-helper cells expressing IL-2 by 68%. GW9662 failed to abrogate this fatty acid effect, likely due to the fact that the compound exerted an own inhibitory effect on IL-2 production. Moreover, GW9662 dose-dependently induced cell death in human leukocytes. These results suggest that application of GW9662 is not conducive in this experimental setting.
authors: Department of Physiology and Biochemistry of Nutrition, Max Rubner-Institut, Federal Research Institute of Nutrition and Food, 76131 Karlsruhe, Germany.
authors: Anke
authors: A
authors: Jaudszus
authors: Department of Nutritional Biochemistry, Institute of Nutrition, Friedrich Schiller University, 07743 Jena, Germany.
authors: Stefan
authors: 0000-0002-9649-840X
authors: S
authors: Lorkowski
authors: Department of Nutritional Physiology, Institute of Nutrition, Friedrich Schiller University, 07743 Jena, Germany ; Food GmbH, 07743 Jena, Germany.
authors: Michael
authors: M
authors: Gruen
authors: Department of Physiology and Biochemistry of Nutrition, Max Rubner-Institut, Federal Research Institute of Nutrition and Food, 76131 Karlsruhe, Germany.
authors: Alexander
authors: A
authors: Roth
authors: Department of Nutritional Physiology, Institute of Nutrition, Friedrich Schiller University, 07743 Jena, Germany.
authors: Gerhard
authors: G
authors: Jahreis