Limited Applicability of GW9662 to Elucidate PPARγ-Mediated Fatty Acid Effects in Primary Human T-Helper Cells

Author:

Jaudszus Anke1,Lorkowski Stefan2ORCID,Gruen Michael34,Roth Alexander1,Jahreis Gerhard3

Affiliation:

1. Department of Physiology and Biochemistry of Nutrition, Max Rubner-Institut, Federal Research Institute of Nutrition and Food, 76131 Karlsruhe, Germany

2. Department of Nutritional Biochemistry, Institute of Nutrition, Friedrich Schiller University, 07743 Jena, Germany

3. Department of Nutritional Physiology, Institute of Nutrition, Friedrich Schiller University, 07743 Jena, Germany

4. Food GmbH, 07743 Jena, Germany

Abstract

Synthetic antagonists of the nuclear receptor PPARγsuch 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 thec9,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/Lc9,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.

Funder

German Research Council

Publisher

Hindawi Limited

Subject

Immunology and Allergy

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