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. Author manuscript; available in PMC: 2022 Apr 1.
Published in final edited form as: Cancer Immunol Res. 2021 Jul 29;9(10):1229–1241. doi: 10.1158/2326-6066.CIR-20-1014

Figure 5. Acquisition of TYRO3 by NK cells enhances NK-cell ex vivo expansion.

Figure 5.

(A)TYRO3− and TYRO3+ NK cells were FACS-sorted and incubated with inactivated APC K562 cells in the presence of IL2 (50 IU/ml) for 7 days, followed by enumerating live NK cells on day 7 (n = 9). (B) TYRO3− and TYRO3+ NK cells were FACS-sorted and cultured in the presence of IL2 (150 IU/ml) for 7 days, followed by enumerating live NK cells on day 7 (n = 4). (C)Primary human NK cells were incubated with inactivated APC K562 cells or APC K562Tyro3 in the presence of IL2 (50 IU/ml) for 7 days, followed by enumerating live NK cells on day 7 (n = 9). (D) Primary human NK cells were incubated with APC K562 cells or APC K562Tyro3 for 1 h. Then, NK cells were separated from tumor cells, followed by co-culture with IL2 (50 IU/ml) for 7 days, followed by enumerating live NK cells on day 7 (n = 3). (E)Primary human NK cells were incubated with APC K562 cells or APC K562Tyro3 for 1 h. Then NK cells were separated from tumor cells, followed by immunoblotting assay. Summary data of p-STAT3Ser727, p-AKT or p-ERK expression are presented after being normalized to β-Actin expression. Each experiment was repeated with NK cells from at least 3 different donors. (F) TYRO3 expression was determined after IL2-stimulated primary human NK cells co-cultured with APC K562ED-Tyro3 cells for 1 h at an E/T ratio of 1:1 (n = 3). (G) Primary human NK cells were incubated with APC K562 cells, APC K562Tyro3 or APC K562ED-Tyro3 in the presence of IL2 (50 IU/ml) for 7 days, followed by enumerating live NK cells on day 7 (n = 5). (H) NK cells in (G) were followed by BrdU assay with 3 different donors. (I) Primary human NK cells were incubated with APC K562 cells, APC K562Tyro3 or APC K562Tyro3_K550A in the presence of IL2 (50 IU/ml) for 7 days, followed by enumerating live NK cells on day 7 (n = 6). (J) TYRO3 expression was determined after IL2-stimulated primary human NK cells were co-cultured with APC K562Tyro3_K550A cells at an E/T ratio of 1:1 (n = 3). Paired t test was used for A, C and E, two-way ANOVA for B and D, one-way ANOVA for G–I. P values were adjusted by the Turkey’s or Holm-Sidak’s method. *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001. Data are presented as mean ± SD.