FIGURE 3.

AMPK and mTOR regulate lipid oxidation and enrich Treg differentiation in vivo. A, CD4⁺ T cells were stimulated with anti-CD3 and anti-CD28 for 18 h, washed, and replated in the presence of anti-CD3 alone, 25 nM rapamycin (Rapa), 2 ng/ml TGFb, 200 μM Etx, or 10 mM FA for an additional 48 h followed by intracellular Foxp3 staining. B, Immunoblot of Th cultures, naive CD4⁺ T cells, and natural Tregs. C–E, Mice were sensitized to OVA in the presence of Met or a PBS followed by aerosol challenge 21 d later and analyzed by intracellular Glut1 staining in CD4⁺ CD44^lowT cells (solid gray) and CD4⁺ CD44^high T cells in the draining lymph node of the lung from mice treated with (black line) or without (gray line) Met (C). In addition, the percentage (D) and number (E) of CD4⁺ Foxp3⁺ cells and total CD4⁺ T cells in the draining lymph node following aerosol challenge were determined. Results are representative of three independent experiments with average and SD determined using the Student t test. *p ≤ 0.05.