Figure 1.

MBP loaded BMDC can prime MBP responsive T cells. BMDC were generated in the presence of 20ng/ml of recombinant GM-CSF for nine days as previously described [11] and activated with 0.1μg/ml LPS and 5ng/ml of GM-CSF for an additional 18 hours. A) Cells were stained for MHC class II, CD80, CD86 expression with (open histogram) or without (grey histogram) LPS maturation and analyzed by FACS. B) Cytokine concentrations in BMDC supernatants were sampled after 18-hour culture with or without LPS. C and D) To study the primary activation of Tg4 T cells, varying numbers (as stated) of AMK₃₅[7] BMDC were cultured with 2x10⁴ CD4⁺ Tg4.CD45.1+ T cells [12] per well. Cell proliferation was assessed by thymidine incorporation. The results are expressed as mean counts per minute ± standard error of the mean. Tg4 T cell production of cytokines (IL-2, IL-17A, GM-CSF, TNF-α and IFN-γ) was assessed in culture supernatants by ELISA. IL-2 was measured in supernatants after 48 hours of culture and IFN-γ, TNF-α, GM-CSF and IL-17A were measured after 72 hours of culture. Data are representative of four independent experiments. BMDC, bone marrow-derived dendritic cells;FACS, fluorescence activated cell sorting; GM-CSF, granulocyte-macrophage colony-stimulating factor; LPS, lipopolysaccharide; MBP, myelin basic protein.