Effector CTL Adhesion Assay

Adhesion of effector CTLs was assessed as described by Matthews and colleagues^{43 (link)}. Briefly, Ibidi VI0.4 m-slides were coated with 6 mg/mL ICAM-1 overnight at 4°C. Effector CD8 T cells at a density of 10⁶ cells/mL in adhesion medium (RPMI with 0.1% BSA, 40 mM HEPES and 2 mM MgCl₂) were injected into a flow system that used a silicone tubing loop connected to a Multi-phaser NE-1000 syringe pump (New Era Pump Systems Inc.). Cells were allowed to flow over the ICAM-1-coated plates at a continuous shear flow rate of 0.5 dynes / cm² for 10 mins. Cells were monitored by microscopy and the number of adherent cells in the field of view was determined at 1 min intervals by manual counting.