Figure 2

MBP loaded BMDC can initiate EAE in mice seeded with MBP-reactive T cells. BMDC were generated from AMK₃₅ or B10.PL mice and matured with LPS as described in Figure 1. A total of 2x10⁶ Tg4.CD45.1 CD4+ T cells was transferred into B10.PLxC57BL/6 mice one day prior to s.c. injection of 2x10⁶ LPS-conditioned AMK₃₅ BMDC (A) or B10.PL BMDC pulsed with 0.1μM MBP Ac1-9(4Tyr) analog peptide (B). of Pertussis toxin was administered (200 ngi.p.) at the time of BMDC transfer. Clinical signs of EAE were assessed daily with the following scoring system: 0, no signs; 1, flaccid tail; 2, impaired righting reflex and/or gait; 3, partial hind limb paralysis; 4, total hind limb paralysis; 5, hind limb paralysis with partial front limb paralysis; 6, moribund or dead. A) Plots show maximal clinical scores of mice administered AMK₃₅ BMDC and the clinical disease curve of three pooled, independent experiments. B) A total of 2x10⁴ Tg4 CD4+ T cells was cultured with LPS-conditioned B10.PL BMDC pulsed with 0.1μM MBP Ac1-9(4Tyr) and assessed for proliferation (thymidine incorporation). Plots show maximal clinical scores of mice administered B10.PL BMDC pulsed with 0.1μM MBP Ac1-9(4Tyr) and the clinical disease curve of three pooled, independent experiments. C) Plot showing timing of disease onset in mice administered AMK₃₅ DC (black line, n=12) and MBP Ac1-9(4Tyr) pulsed DC (dashed line, n=20). All mice were bred under specific pathogen-free conditions at the University of Edinburgh and all experiments had local ethical approval and were performed in accordance with UK legislation. BMDC, bone marrow-derived dendritic cells; DC, dendritic cells; EAE, experimental autoimmune encephalomyelitis; LPS, lipopolysaccharide; MBP, myelin basic protein.