Fig. 4. LOX pathways play a role in BCG-induced trained immunity.

a Production of TNFα and IL-6 by BCG-trained macrophages incubated with (10, 100 µM) Zileuton, (2, 20 µM) ML355 or 10 µM PD146176 for the first 24 h and restimulated with LPS for another 24 h at day 6 of culture. (n = 9 biologic replicates, pooled from 3 independent experiments). b–d Production of TNFα and IL-6 by macrophages incubated with b (3, 30, 300 nM) 5-HETE, (0.1, 1, 5 µM) 7-HDHA, (1, 10 nM) LTB₄, (c) (0.03, 0.3, 3 µM) 15-HETE, (0.1, 1, 5 µM) 17-HDHA, d (0.1, 10 nM) 12-HETE (0.1, 10 nM) 12-HEPE or (0.1, 10 nM) 14-HDHA for 24 h and restimulated at day 6 of culture with LPS for 24 h. (n = 6 (b, c) or n = 9 (d) biologic replicates, pooled from 2 (b, c) or 3 (d) independent experiments). Mean + SD, a two-way ANOVA, Sidak’s multiple comparisons test, b–d Friedman test followed by Dunn’s multiple comparisons correction. (Ctrl: control; BCG: Bacillus Calmette-Guérin; LPS: lipopolysaccharide; LOX: lipoxygenase).