Muramyl dipeptide activation of nucleotide-binding oligomerization domain 2 protects mice from experimental colitis
J. Clin. Invest. Tomohiro Watanabe, et al. 118:545 doi:10.1172/JCI33145 [Go to this article.]

Figure 2
MDP administration reduces the TLR-induced cytokine responses of MLN and colonic LP cells from mice with TNBS colitis. (A) Colon LP lymphocytes (1 × 10⁶/ml) isolated from the mice on day 3 were stimulated with PGN, Pam₃CSK4, LPS, and CpG in the presence of IFN-γ (20 ng/ml); cultured supernatants were collected at 48 hours and analyzed for cytokine production. *P < 0.05; **P < 0.01 when supernatants from MDP-treated mice were compared with supernatants from PBS-treated mice. (B) MLN cells (1 × 10⁶/ml) isolated from mice with TNBS colitis on day 3 were stimulated with anti-CD3 (1 μg/ml) and a broad range of TLR ligands. Cultured supernatants were collected at 48 hours and analyzed for cytokine production. *P < 0.05; **P < 0.01 when supernatants of MDP-treated mice were compared with supernatants of PBS-treated mice. (C) MLN cells and colon LP lymphocytes isolated from the mice on day 3 were stimulated with anti-CD3 (1 μg/ml); cultured supernatants were collected at 48 hours and analyzed for IFN-γ production. **P < 0.01 when supernatants from MDP-treated mice were compared with supernatants from PBS-treated mice. (D) Evaluation of NF-κB activation in MLN cells isolated from the mice on day 3 and stimulated with LPS or PGN. Nuclear extracts were prepared from MLN cells isolated from PBS- or MDP-treated mice and stimulated with LPS or PGN for 2 hours and then subjected to EMSA. (E) Nuclear extracts obtained in D assayed for p65 and c-Rel activation using NF-κB transcription factor ELISA. *P < 0.05; **P < 0.01 compared with nuclear extracts from PBS-treated mice.