LNCs from MOG
-immunized WT, Astro KO , and Endo KO mice show similar responses to MOG
restimulation in vitro . LNCs were prepared from MOG35-55-immunized mice on d12, and restimulated with MOG35-55 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 MOG38-49 MHC class II tetramer-PE. Binding was determined after 72 h in culture, and hCLIP103-117 tetramer-PE served as a control for non-specific binding. Plots were gated on CD4+ T lymphocytes. The frequency of MOG38-49 I-Ab tetramer+ CD4+ T cells is similar among WT, Astro KO, and Endo KO groups, while hCLIP103-117 I-Ab 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.