Fig 6. TLR3, 7 and 9 upstream of MyD88 extrinsically regulate T cell contraction.

One million CD4⁺ T cells from naïve 1807 mice were transferred into wild type mice prior to vaccination. Seven days post-vaccination, effector CD4⁺ T cells were magnetic bead-purified from the sdLN and spleen of the vaccinated mice and adoptively transferred into naïve Myd88 ^-/-, IL-1R^-/-, TLR23479^-/-, TLR379^-/- and TLR24^-/- and wild type mice. After 4 weeks of rest, primed 1807 T cells harvested from the lymph nodes and spleen were enumerated by FACS. (A) Data are expressed as the mean ± SD of 8–12 mice/group. Data represent the average from two independent experiments. * P < 0.05 vs. wild type control mice. (B) shows the dot plots of concatenated events from 8–12 mice/group from panel A. * P < 0.05 vs. wild type control mice. (C) Myd88 ^-/-, IL-1R^-/-, TLR23479^-/-, TLR379^-/- and wild type mice were vaccinated with 10⁶ heat-killed vaccine yeast. At days 7 and 49 post-vaccination, the number of Cnx-specific CD4⁺ T cells in the sdLN and spleen was enumerated using tetramer enrichment and FACS detection. Tetramer positive cells are shown within the gate in each dot plot. The numbers represent the geometric mean ± SEM of tetramer-positive cells of 5 mice. Data are expressed from a single experiment representative of three independent experiments. * P < 0.05 vs. wild type control mice. (D) Myd88 ^-/-, IL-1R^-/-, TLR23479^-/-, TLR379^-/-, TLR24^-/- and wild type mice were vaccinated as above, challenged and lung CFU enumerated at day 14 post-infection when unvaccinated mice were moribund. The numbers indicate the n-fold change vs. unvaccinated mice. Data are the mean ± SD of 10–20 mice/group from a single experiment representative of three independent experiments. * P < 0.05 vs. vaccinated wild type control mice.