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. 2013 Oct 15;21(12):2268–2277. doi: 10.1038/mt.2013.192

Figure 3.

Figure 3

CD56+ cytokine-induced killer (CIK) cells are terminally differentiated T cells that are induced by OX40 costimulation and prone to activation-induced cell death. (a) CIK cells were generated as described in Materials and Methods. After 14 days of culture, cells were separated into CD56+ and CD56− cells and stained for CD45RO and CD62L, respectively. A representative experiment out of three is shown. (b) CIK cells from healthy donors were grafted with the ζ (n = 5), CD28-ζ (n = 12), or CD28-ζ–OX40 CAR (n = 12) and cultivated for 3 days in the presence of the anti-idiotypic monoclonal antibody (mAb) BW2064 (1 µg/ml) and an antimouse immunoglobulin G1 (IgG1) (0.5 µg/ml) antibody for CAR crosslinking. Cells were stained for CD56 and CAR expression and analyzed by flow cytometry. Nonmodified CIK cells were stained for control (n = 4). Numbers represent the mean value ± SD. Statistical analyses were performed using Student's t-test. (c) CIK cells from six healthy donors were engineered with the CD28-ζ or CD28-ζ–OX40 CAR and cultivated for 3 days in the presence of the anti-idiotypic mAb BW2064 (1 µg/ml) and an antimouse IgG1 (0.5 µg/ml) antibody for CAR crosslinking. Cells were stained for CD56 and CAR expression and apoptotic cells were identified by annexin V expression. Numbers represent mean value ± SD. P values were determined by Student's t-test. CAR, chimeric antigen receptor.