Figure S1. Immunological characterization of BM-DC used for immunization. (a) CD11c expression, measured at days 1, 4, 7, 10 and 14 of BM culture by flow cytometry (FACS) in three independent experiments, demonstrating a peak of 80–90% at day 10 of culture. (b) DC-45D, as prepared for immunization, was characterized for surface marker expression by FACS analysis, as compared to DC-45D activated with LPS (LPS DC-45D). Out of total CD11c+ cells in the cell preparation used for immunization, 8.3% expressed CD83, 33.2% expressed B7-1 (CD80), 23.5% expressed B7-2 (CD86), and 7.8% expressed CD40. Results are expressed as % ± SD. According to the cell surface profile used to characterize the DCs, no significant differences were detected between the DC-45D and DC (data not shown). Secretion levels of pro-inflammatory cytokine in DC-45D, such as interleukin (IL)-12, IL-6, IL-1α and TNFα, measured by ELISA, were significantly lower than those obtained from LPS DC-45D (data not shown). (c and d) DC-45D evoked a specific immune response in vivo, assessed by a lymphocyte proliferation assay using CFSE staining (see Methods). Mice were immunized with either DC-45D or with peptide-free DCs (Muir et al.). After 6 days, lymph nodes were tested ex vivo for proliferation in the presence of either MOG45D (the relevant neuropeptide that was injected in vivo) or Nogo-A (an irrelevant neuropeptide as control). (c) Significant proliferation of CD3+ T cells (= 0.024) and (d) CD19+ B cells (= 0.04) was observed specifically in DC-45D-immunized mice relative to DC-immunized mice in response to MOG45D (index, ± SD). Asterisks indicate statistical significance analyzed by the Student’s t-test: *< 0.05, **< 0.005, ***< 0.0001.

Figure S2. No signs of demyelination were found following DC-immunization in AD-Tg mice. Representative microscope images show two myelin-rich areas in the brain, corpus callosum and striatum, stained with (a–d) anti-MBP, and (e–h) Luxol fast blue, with no detectable demyelination. Scale bar scale indicates 100 μm.

As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer-reviewed and may be re-organized for online delivery, but are not copy-edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.

JNC_6402_sm_Figure S1.eps345KSupporting info item
JNC_6402_sm_Figure S2.eps10488KSupporting info item
JNC_6402_sm_Supplementary Figures.doc56KSupporting info item

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.