FIGURE 2.

CD80/CD28 interactions prominently regulate CD8⁺ T-cells, while PD-1/PD-L2 control the CD4⁺ T-cell responses in WT mice subjected to MCAO. T-cells were purified, by negative sort, from the spleens of MCAO-subjected WT mice, by labeling with specific microbeads and separating on the AutoMACS^TM. The purified T-cells were CSFE-labeled and then cultured with non-T-cells from the same mice at a 1:1 ratio (T:APC) in the presence of anti-CD3 antibody (2.5 μg/mL; baseline) and other neutralizing Abs (10 μg/mL) to co-stimulatory molecules, in 96-well plates. After 72 h of culture, cells were washed and evaluated by FACS Calibur for (A) CD8⁺ and (B) CD4⁺ T-cell expression and CFSE dilution. Data represent the stimulation indices of the CD8 and CD4 T-cells in the presence of neutralizing Abs as compared to the control (T:APC + anti-CD3 Ab) condition). The data are represented such that the baseline value is 1 and all other values are adjusted relative to the baseline. Data presented are representative of splenocytes obtained from nine WT mice, with least three separate experiments and each experiment comprising duplicates or triplicates of the given neutralizing Ab condition. Significant differences between sample means are indicated as *p ≤ 0.05, **p ≤ 0.01 as compared to the baseline condition.