Figure 2.

Unlike those from healthy individuals, OCs from cancer patients induced decreased cell expansion, IFN-γ secretion and cytotoxicity in allogeneic NK cells obtained from healthy individuals. NK cells (1 × 10⁶ cells/ml) from healthy individuals were treated with the combination of IL-2 (1000 U/ml) and anti-CD16mAb (3 µg/ml) for 18 h before they were cultured alone or were co-cultured with either healthy individuals’ OCs or cancer patients’ OCs in the presence of sAJ2 at a ratio of 1:2:4 (OCs:NK:sAJ2). On days 6, 9, 12, 15, 18 and 22 of co-culture, the numbers of NK cells were counted using microscopy (n = 12) (A). NK cells were treated and co-cultured as described in Fig. 2A. On days 6, 9, 12, 15, 18 and 22, supernatants were harvested from the co-cultures to determine IFN-γ secretion using single ELISA (n = 12) (B). The amounts of IFN-γ secretion shown in Fig. 2B were assessed based on 1 × 10⁶ cells (n = 12) (C). NK cells were treated and co-cultured as described in Fig. 2A. Cytotoxicity of days 9 and 15 cultured NK cells were determined using a standard 4-h ⁵¹Cr release assay against OSCSCs. LU 30/10⁶ cells were determined as described in Fig. 1B (n = 4) (D). NK cells (1 × 10⁶ cells/ml) from healthy individuals and cancer patients were treated with the combination of IL-2 (1000 U/ml) and anti-CD16mAb (3 µg/ml) for 18 h before they were cultured alone, or with autologous OCs in the presence of sAJ2 at a ratio of 1:2:4 (OCs:NK:sAJ2). On days 6, 9, 12, and 16 of co-culture, the numbers of NK cells were counted using microscopy (E). NK cells were treated and co-cultured as described in Fig. 2E. Cytotoxicity of days 9 and 15 cultured NK cells were determined using a standard 4-h ⁵¹Cr release assay against OSCSCs. LU 30/10⁶ cells were determined as described in Fig. 1B (n = 4) (F). NK cells were treated and co-cultured as described in Fig. 2E. On days 6 and 12, supernatants were harvested from the co-cultures to determine IFN-γ secretion using single ELISA (G).