Anti-TNF-α Therapy Suppresses Proinflammatory Activities of Mucosal Neutrophils in Inflammatory Bowel Disease

Figure 4

Anti-TNF-α therapy inhibits peripheral neutrophils to produce proinflammatory cytokines, chemokines, ROS, and MPO. Peripheral neutrophils (5 × 10⁶) from healthy donors (), patients with active CD (), and active UC () were stimulated by LPS (200 ng/ml) and incubated with or without IFX (50 μg/ml) for 3 h. Cells were collected and expressions of TNF-α (a), IL-8 (b), and IL-6 (c) were detected by qRT-PCR. Peripheral neutrophils (2 × 10⁶) from healthy donors (), patients with active CD (), and patients with active UC () were simulated with LPS (200 ng/ml) and incubated with or without IFX (50 μg/ml) for 3 h. Culture media were replenished and incubated for another 24 h. Supernatants and protein production of TNF-α (d), IL-8 (e), and IL-6 (f) were measured by ELISA. Peripheral neutrophils (1 × 10⁴) isolated from healthy donors (), patients with active CD (), and patients with active UC () were measured for ROS (h) and MPO (i) with Amplex Red Hydrogen Peroxide Assay Kit. , , and compared with medium control and , , and compared with LPS stimulation.

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)