Expression of proinflammatory cytokines and early TNFα and IL-10 secretion are restricted to a subset of TLRs.
A, BMDMs from WT and tpl2−/− mice were either left untreated or stimulated with Pam3CSK4, poly I:C, LPS, R848, or CpG ODN for 1 h. Cells were lysed after collecting supernatants, and il12b, tnfa, il10, and ccl2 mRNA expression were measured by real-time PCR relative to an actin control. B, TNFα and IL-10 secretion at 1 h were measured by ELISA. Values are relative to WT cells treated with LPS, which was considered 100% in individual experiments. C, BMDMs from WT and tpl2−/− mice were either left untreated or stimulated with poly I:C, LPS, or CpG ODN for 1, 2, 4, or 24 h. TNFα levels in cell culture supernatants were measured by ELISA. ** indicates p < 0.01, * indicates p < 0.05. For A-C, bars indicate mean ± S.E. from 3–4 independent experiments.