Fig. 8.

4-TMPs activate DCs regardless of TCR engagement. a Representative confocal images of Fluo-3-loaded DCs placed on a 4-TMP-deposited lipid bilayer. b Fluo-3-loaded DCs from wild type or TLR4^−/− mice were stimulated with 4-TMPs purified from CD4⁺ T cells in the presence or absence of SEB. Calcium changes were analysed by flow cytometry for 300 s. Data are representative of at least three separate experiments. c Surface protein expression on DCs induced by 4-TMPs. DCs (5 × 10⁶) from wild-type mice were treated with 4-TMPs obtained from CD4⁺ T cells (0.05–1 × 10⁸) at the indicated ratio (DC:T = 1:1 to 1:20). After 24 h, the expression of CD86 was analysed by flow cytometry and expressed as the mean fluorescence intensity (MFI). Data represent the mean of three experiments ± SEM. *P < 0.01 vs. 1:0. d DCs (5 × 10⁶) from wild type or TLR4^−/− mice were treated with 4-TMPs (5 × 10⁷). After 24 h, the expression of CD86, CD40, and CD80 was determined as described in c. In some cases, FcγR was blocked 30 min before 4-TMP treatment. *P < 0.01 vs. PBS-treated group