Signaling through the chNKG2D receptor decreases expression of anti-inflammatory cytokines and a proangiogenic molecule, VEGFα. chNKG2D T cells were stimulated with media (gray bars), anti-CD3/CD28–coated (filled bars), anti-CD3– (hashed bars), or anti-NKG2D–coated (open bars) beads for 24 hours. Expression of GM-CSF, IFNγ, TNFα, and IL-2 (A-B) or IL-9, IL-13, IL-10, and VEGFα (C-D) was determined by RT-PCR (A,C) or by measuring levels in cell-free supernatants by Luminex analysis (B,D). RT-PCR data are shown as the fold change in expression compared with T cells cultured in media and are representative of data from ≥ 8 donors. Cytokine secretion data are presented as mean ± SD and are from 2 donors (as indicated by donors 6 and 7). Similar cytokine secretion data were obtained when 72-hour supernatant fluids were analyzed from 2 additional donors (data not shown). When stimulated through NKG2D, chNKG2D T cells produced more inflammatory cytokines and less anti-inflammatory cytokines and VEGFα than when stimulated through CD3 alone.