Ghrelin inhibits leptin- and activation-induced proinflammatory cytokine expression by human monocytes and T cells
J. Clin. Invest. Vishwa Deep Dixit, et al. 114:57 doi:10.1172/JCI21134 [Go to this article.]

Figure 1
Expression of functional GHS-R in human T cells. (A) Primary human T cells were labeled for GHS-R, and subcellular localization in lipid rafts was visualized in resting and anti-CD3–activated cells. Arrowheads indicate colocalization of GHS-R with polarized lipid rafts. (B) Flow cytometric analysis of GHS-R on highly purified resting human T cells. Specific T cell labeling (green) was abolished in presence of antibody-specific blocking peptide (blue). T cells stained with control IgG demonstrated no specific labeling. (C) Flow analysis of GHS-R expression on highly purified (>96%) activated CD3⁺ human T cells. Staining specificity was demonstrated through the use of antibody-specific blocking peptide. (D) GHS-R mRNA is upregulated upon T cell activation as assessed using Agilent gene chip quantitation and real time RT-PCR; values are expressed as mean ± SEM (*P < 0.05). (E) Ghrelin induces intra-cellular calcium mobilization in cultured human T cells. T cells were stimulated with ghrelin (100 ng/ml; blue), or SDF-1 (100 ng/ml; green) at 60 seconds. T cells were also treated with the GHS-R antagonist, [D-Lys-3]-GHRP-6 (10^–4 M), at 60 seconds followed by ghrelin (100 ng/ml; red) at 180 seconds. FL, fluorescence. (F) Ghrelin causes actin-polymerization in human T cells. Cells were treated with ghrelin (100 ng/ml) and positive control SDF-1α (100 ng/ml) for 20 minutes and labeled for F-actin with phalloidin AF-594. Arrowheads indicate polymerized F-actin associated with lamellipodia of cells.