Figure 4

LNCs from MOG _35-55 -immunized WT, Astro KO , and Endo KO mice show similar responses to MOG _35-55 restimulation in vitro . LNCs were prepared from MOG_35-55-immunized mice on d12, and restimulated with MOG_35-55 for 72 h in culture, after which time different responses were measured. (a) T cell proliferation. LNC were pulse-labeled with 2 μΜ CFSE for 5 min at the beginning of culture and analyzed after 72 h by FACS, gating on CD3, CD4, CD11a. (b) Cytokine production. The concentrations of IL-17 and IFN-γ were determined in supernatants of LNC after 72 h in culture. (c) Binding of MOG_38-49 MHC class II tetramer-PE. Binding was determined after 72 h in culture, and hCLIP_103-117 tetramer-PE served as a control for non-specific binding. Plots were gated on CD4⁺ T lymphocytes. The frequency of MOG_38-49 I-A^b tetramer⁺ CD4⁺ T cells is similar among WT, Astro KO, and Endo KO groups, while hCLIP_103-117 I-A^b tetramer does not bind cultured T cells. The data shown are representative of at least two independent experiments; data in (a) and (b) reflect mean value ± standard error.