Clone cd3 12

Mice were transcardially perfused with PBS containing 5 IU/ml heparin (De Pannemaeker N.V., Gent, Belgium) followed by perfusion with 4% paraformaldehyde. Spinal cords were dissected, dehydrated and embedded in paraffin blocks. Sections of 2 μm were stained with hematoxylin and eosin (H & E), Luxol fast blue (LFB) (Solvent Blue 38, practical grade, Sigma Genosys, The Woodlands, TX, USA) for assessment of demyelination, and antibodies against CD3 (Clone CD3-12, Serotec, Raleigh, NC, USA), Mac-3 (Clone CD107b, M3/84, BD Biosciences, San Diego, CA, USA), B220 (Clone RA3-6B2, BD Biosciences, San Diego, CA, USA) or amyloid precursor protein (APP) (Clone 22C11, Millipore, Darmstadt, Germany). Sections were rehydrated and incubated in 10 mM citrate buffer for 5 minutes at 94°C. Nonspecific binding was blocked by incubating sections in 0.1 M PBS containing 10% FCS and 1% Triton x-100 for 30 minutes. Primary antibodies were incubated overnight at 4°C. Histological quantification was described previously [18 (link)].