Skip to main content
Fig. 3 | Journal of Inflammation

Fig. 3

From: Prediction and identification of novel HLA-A*0201-restricted cytotoxic T lymphocyte epitopes from endocan

Fig. 3

Specific lysis of endocan-derived peptide elicited CTLs. The PBMCs of human HLA-A2+ donors were obtained and then cultured in RPMI 1640 supplemented with 10% FCS, 100 U/ml penicillin, and 100 μg/ml streptomycin. DCs were generated, and loaded with different peptides at a final concentration of 100 μg/ml for 4 h and were then irradiated with 20 Gy, which prevented all outgrowths in the control cultures. Autologous T cells were restimulated every 7 days with the peptide-pulsed DCs to generate peptide-specific CTLs. Target cells were incubated with 51 Cr (100 μCi per 1× 106 cells; Amersham Biosciences Corp) for 2 h in a 37 °C water bath. After incubation with 51Cr, target cells were washed three times with PBS, resuspended in RPMI 1640, and mixed with effector cells at effector-to-target (E/T) ratios of 25:1, 50:1, or 100:1. The percentage of cytotoxicity was calculated as follows: percentage of lysis = (sample cpm- spontaneous cpm)/(maximum cpm- spontaneous cpm) × 100%. Experiments performed in triplicate showed consistent results. Compared with control, *P < 0.05

Back to article page