Assay for CD8+ T Cell Adhesion

Cellular adhesion was performed as previously described with some modification (Bilal et al., 2015 (link); Chapman et al., 2012 (link)). Briefly, flat-bottomed 96-well plates (Thermo-Fisher) were coated with 0–10 μg of αCD3 (OKT3, Biolegend). P14 CD8 T cells were isolated by positive selection, based on Thy1.1. 5 × 10⁶ P14 CD8 T cells were incubated on the plate for 30 min. Non-adherent cells were removed by quickly inverting the plate to empty contents. Adherent cells were stained with αCD8a-APC-Cy7 (53–6.7; Biolegend). Cells were washed twice with PBS before being imaged utilizing Licor Odyssey Infrared detector.