Article Figures & Data
Figures
- FIG 1
Schematic overview of trained immunity methodology. Monocytes were trained for 2 h (2 hr-T), 4 h (4 hr-T), or 24 h (24 hr-T). After the training stimulus was washed away, the cells were rested for 24 h (24 h-R), 3 days (3d-R), or 6 days (6d-R), after which the cells were restimulated with RPMI, LPS, or Pam3Cys for 24 h.
- FIG 2
Increased proinflammatory cytokine production is dependent on both the training interval and the resting time. (A) IL-6 production after restimulation. Cells were trained for 2 h, 4 h, or 24 h with β-glucan, BCG, or oxLDL and rested for 24 h, 3 days (3d), or 6 days (6d). (B) TNF-α production after restimulation. Cells were trained for 2 h, 4 h, or 24 h with β-glucan, BCG, or oxLDL and rested for 24 h, 3 days, or 6 days. Shown are the fold changes compared to the results for the RPMI control (n = 6; ^, P = 0.06 compared to the RPMI control; *, P < 0.05 compared to the RPMI control; **, P < 0.01 compared to the RPMI control).
- FIG 3
Trained immunity effects on cell morphology and numbers. (A) Morphology of cells after 24 h of training and 6 days of rest when the cells were trained with RPMI (negative control), β-glucan, BCG, or oxLDL. Pictures were taken before restimulation at day 6. Magnification, ×20. (B) Size of cells after 24 h of training and 6 days of rest before restimulation at day 6 (n = 3, *, P < 0.05; compared to the RPMI control; ***, P < 0.001 compared to the RPMI control). (C) Relative cell counts before restimulation at day 6 (n = 6; *, P < 0.05, compared to the RPMI control; ns, not significant). (D) IL-6 and TNF-α production after restimulation for corrected cell counts (n = 6; *, P < 0.05 compared to the RPMI control; **, P < 0.01 compared to the RPMI control).
- FIG 4
Anti-inflammatory cytokine production is increased in trained monocytes. (A) IL-10 production after restimulation. Cells were trained for 24 h with β-glucan, BCG, or oxLDL and rested for 6 days. The level of production of the anti-inflammatory cytokine IL-10 increased upon training. (B) IL-1Ra production. Cells were trained for 24 h with β-glucan, BCG, or oxLDL and rested for 6 days. The level of production of the anti-inflammatory cytokine IL-1Ra increased upon training not only after restimulation but also at the baseline (n = 6; ^, P = 0.06 compared to the RPMI control; *, P < 0.05 compared to the RPMI control; **, P < 0.01 compared to the RPMI control; ns, not significant). P3C, Pam3Cys.
- FIG 5
ROS production is a component of trained immunity for some stimuli. The level of ROS production was significantly increased for monocytes trained with BCG and oxLDL; training with β-glucan decreased the level of ROS production (n = 6; *, P < 0.05 compared to the RPMI control; ***, P < 0.001 compared to the RPMI control; ns, not significant). Cells were trained for 24 h using β-glucan, BCG, or oxLDL, and normalized cell numbers were restimulated using zymosan. Luminol was added, and luminescence was measured for 1 h. RLU/s, relative light units per second.
- FIG 6
Trained immunity induction of glycolysis. Cells were trained for 2 h (2hr-T), 4 h (4hr-T), or 24 h (24hr-T) with RPMI (negative control), β-glucan, BCG, or oxLDL and rested for 24 h (24hr-R), 3 days (3d-R), or 6 days (6d-R). The level of lactate production in the supernatants was measured before restimulation (n = 6; *, P < 0.05 compared to the RPMI control; **, P < 0.01 compared to the RPMI control; ns, not significant).
Additional Files
Supplemental material
- Supplemental file 1 -
Fig. S1. Monocyte purity of different isolation steps. Fig. S2. Increased proinflammatory cytokine production is dependent on both training and resting time. Fig. S3. Training induces changes in cell morphology. Fig. S4. Anti-inflammatory cytokine production is increased in trained monocytes. Fig. S5. Morphological changes after 24 h of training and 6 days of resting persist after 24 h of restimulation.
PDF, 2.5M
- Supplemental file 1 -
