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. 2019 Nov 19;10:2674. doi: 10.3389/fimmu.2019.02674

Figure 4.

Figure 4

Macrophages differentiated with IFN-λ3 induce lymphocyte chemotaxis and NK cell activation. Transwell inserts containing autologous PBMCs were placed into wells containing 6-day differentiated Mϕs ± IFN-λ3 to examine immune cell chemotaxis (n = 6). Migrated cells were analyzed by flow cytometry using CD14, CD19, CD3, and CD56 antibodies to identify the number of migrated monocytes, B, NK, and T cells (A). The addition of IFN-λ3 to GM-CSF Mϕs alone, stimulated lymphocyte migration, with significant increases in NK, NKT, and T cell migration (B). Consistent with migration data, GM-Mϕ CCL2, CCL3,CCL4, and CXCL10 secretion were significantly increased by IFN-λ3, with no significant effect on M-Mϕs (C) (n = 7). To assess the role of IFN-λ3 on Mϕ activation of NK cells, autologous NK cells were incubated with Mϕs for 16 h a ratio of 1:1, removed and incubated with K562 target cells for a further 6 h to measure cytotoxicity by degranulation and IFN-γ expression (n = 4). GM-Mϕs differentiated with IFN-λ3 significantly increased NK cell cytotoxicity as measured by CD107a expression, as well as IFN-γ production (D). Data are representative of two (B,D) and three (C) independent experiments. Paired t-test *p < 0.05, **p < 0.01 (mean ± SE). NE, No expression.