Anti cd11a m17 4

All animal work was approved by the Institutional Animal Care and Use Committee of the University of California, San Diego. All mice were housed in specific pathogen-free conditions prior to use. Cd4^Cre mice were purchased from Jackson Laboratories, and Tln1^fl/fl and Foxp3^GFP mice have been described previously (6 (link), 32 (link)–34 (link)). For administration of IL-2/IL-2mAb complexes, 2μg of recombinant murine IL-2 (Biolegend) was combined with 10μg IL-2 monoclonal antibody JES6-1 (Bio X Cell), diluted to a volume of 200μL in PBS and incubated for 30 minutes at 37°C before intraperitoneal (i.p.) injection, as previously described (35 (link), 36 (link)). To block LFA-1 signaling, mice were treated with 100μg anti-CD11a (M17/4) (Bio X Cell), 100μg anti-CD18 (M18/2) (Bio X Cell) and 200μg anti-ICAM-1 (YN1/1.7.4) or isotype control in PBS via i.p. injection twice weekly for three weeks.

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.