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. 2021 Aug 31;12:733136. doi: 10.3389/fimmu.2021.733136

Figure 4.

Figure 4

Blocking PD-1 on ILC2s increases TNF-α production and enhances cytotoxic properties. (A) A cohort of WT mice was given an intravenous injection of B16 melanoma on day 0. On day 14 lung ILC2s were sorted by flow cytometry and cultured with isotype or PD-1 blocking antibody for 48 hours. (B) After 48 hours, ILC2 cells were harvested and measured for TNF-α expression by intracellular flow cytometry. Representative flow cytometry plots and corresponding quantification are presented as percent TNF-α+ ILC2s. (C) B16 melanoma cells were cultured in the bottom well of 0.4 µm Transwell plate. PD-1-/- or WT ILC2s were cultured in the top well insert at 8:1 ratio with B16 melanoma cells respectively for 48 hours. In the condition specified, anti-TNF-α or isotype (10µg/ml) was included in the bottom well of the culture. After 48 hours, B16 cells were collected and stained with apoptosis kit to assess viability. Representative flow cytometry plots and corresponding quantification are presented. Error bars are the mean ± SEM. Data are representative of 3 individual experiments with n=5. Student’s t-test, **p < 0.01, ***p < 0.001.