Fig. 4.

Neutralization of leptin decreases the percentage of T helper type 17 (Th17) cells in vitro. Human CD4⁺ T cells were purified from peripheral blood mononuclear cells (PBMCs) by magnetic beads and cultured with RPMI-1640 medium supplemented with 10% fetal bovine serum; 10 μg/ml human leptin-neutralizing monoclonal antibody (mAb) was administered in CD4⁺ T cell culture in the presence of soluble anti-human CD3 mAb and anti-human CD28 mAb; the irrelevant isotype-matched antibody was used as control. The percentages of CD4⁺ Th17 cells were analysed by flow cytometry (a); retinoic acid-related orphan receptor γt (RORγt) mRNA were determined by quantitative PCR (b).