Fig. 3
ADSC are suppressive against NK cells and can overcome allogeneic immune barriers. a ADSC suppress NK cells in autologous and allogeneic settings. Freshly isolated RM20 PBMCs (250,000) were cocultured for 48 h with 10, 000 or 100,000 autologous (RM20) or allogeneic (RM35) ADSC. To evaluate the extend of NK suppression the 48 h cocultures were subjected to an additional 4 h stimulation of NK cells with 250,000 K562 cells or PMA/Ionomycin. Data represent flow cytometry analysis of CD69 upregulation on gated live CD3-NKp46 + NK cells. b Flow cytometry analysis of NK cell cytotoxic functions using CD107a surface exposure as in a. Bars represent triplicate measurements ± SD. Statistically significant differences (Student T-test, p < 0.05) versus the corresponding (−) ADSC (PBMC alone) controls are marked with asterisks (c) ADSCs can amplify vaccinia virus in the presence of allogeneic PBMC. RM20-eGFP ADSCs (50,000) were infected with 5000 or 50,000 L14 VV alone or in the presence of 1 × 106 allogeneic PBMC (BH062 blood donor) for up to 48 h. Overlay fluorescence imaging (top) and plaque assay (bottom) were used to evaluate vaccinia virus infection and amplification, respectively. Statistically significant differences (Student T-test, p < 0.05) based on duplicates ± SD are marked with asterisks