Anti ccr7 4b12

Naïve OT-I T-cells isolated from pooled LN and spleen of OT-I RAG1^−/−/Thy1.1 mice (4×10⁶ cells) or polyclonal CD8⁺ T-cells pooled from LN and spleen of C57BL/6 mice and depleted of CD44 expressing cells by magnetic beads (Miltenyi) were injected into the lateral tail vein of recipients. In blocking experiments, cells were incubated with 100 ng/mL pertussis toxin (PTX, Sigma) for 1 h at 37°C, or 100 µg of rat IgG (Jackson Immunoresearch), anti-CD62L (Mel-14, ATCC), or anti-CD11a (M17/4, BioXcell), or 50 µg of anti-CCR7 (4B12, eBioscience) and washed before injection. Luminal PNAd or MAdCAM-1 were blocked by injection of 100 µg of anti-MAdCAM-1 (MECA-367, BioXcell), anti-PNAd (MECA-79, BioLegend) or isotype rat IgG or IgM i.v. 1 h prior to T-cell transfer. In some experiments, cells were labeled with Cell Trace Violet (CTV) (Invitrogen) prior to transfer. Tissues were harvested 1 h or 18 h after naïve T-cell transfer.