Briefly, single - cell suspensions of C57BL / 6 KO and wild - type lymphocytes, which had undergone a 96 hr in vitro boost with γ -
irradiated (3,000 rad)
BALB / c stimulator lymphocytes, were added to 96 - well microtiter plates (Corning Inc., Corning, NY) along with 2 × 104 51Cr - labeled
BALB / c corneal epithelial or endothelial cells in a total volume of 200 μl / well.
Spleen cells (3 × 107) from either experimental or control effector lymphocytes were dispensed into 75 - cm2 tissue culture flasks along with 20 ml of boosting medium and 6 × 105 γ -
irradiated (3,000 rad)
BALB / c spleen cells (erythrocytes lysed).