Extended Data Fig. 7: T_β-syn cell activation drives immune cell recruitment.

a, Recruitment of host T cells and myeloid cells determined by flow cytometry at the peak of disease induced by T_β-syn cell transfer. Data are mean + s.e.m., n = 3. b–d, T_β-syn cell invasion in the CNS drives recruitment of CNS-ignorant T_OVA cells. b, T_OVA cell recruitment to the grey matter cortex after transfer of T_OVA–Tomato cells alone (top) or after cotransfer with T_{β-syn–Turquoise} cells (bottom). Left, fluorescence overview images and magnified views of selected areas of coronal brain sections. Right, TPLSM individual stacks and corresponding migratory paths of T_β-syn and/or T_OVA cells over a 30-min recording time. Data were acquired 3.5 days after T cell transfer. Arrows point to representative T_OVA cells. Red, T_OVA cells; turquoise, T_β-syn cells, green, blood vessels (labelled by FITC-conjugated dextran). Representative data of at least three independent experiments (obtained from individual rats). c, Time course of T cell infiltration in the indicated compartments upon single transfer of T_OVA–Tomato cells or cotransfer of T_{β-syn–Turquoise} and T_OVA–Tomato cells, determined by flow cytometry. Data are mean + s.e.m., n = 3 per time point. Representative data of two independent experiments. d, T_OVA cells are not reactivated in the brain. Expression level of the indicated cytokines measured by quantitative PCR in T_β-syn and T_OVA cells isolated from the indicated compartments upon cotransfer. Data are mean + s.e.m., n = 3. Representative data of two independent experiments. e, FK506 treatment prevents T_OVA-cell recruitment to the grey matter tissue. Fluorescence overview images as in b from control (clinical score 2) or FK506-treated (clinical score 0) rats 3.5 days after cotransfer. Representative images of two independent experiments including three rats per group. f, Blockade of antigen presentation by intrathecal administration of anti-MHCII monoclonal antibodies effectively reduces immune cell recruitment to the brain in the cotransfer experiment. Left, ratio of the numbers of T_β-syn or T_OVA cells in brain tissue to the number of T_β-syn or T_OVA cells in blood. Data are mean + s.e.m., n = 3. Middle and right, absolute number of myeloid cells or T cells in the brain tissue measured by flow cytometry 3.5 days after cotransfer of T_β-syn and T_OVA cells. Data are mean + s.e.m., n = 3. f, Unpaired two-tailed t-test. *P < 0.05.