Interleukin-36γ and IL-36 receptor signaling mediate impaired host immunity and lung injury in cytotoxic Pseudomonas aeruginosa pulmonary infection: Role of prostaglandin E2
Fig 2
P. aeruginosa induced IL-36α and IL-36γ expression in primary pulmonary macrophages and alveolar epithelial cells.
Primary pulmonary macrophages (PMs) and alveolar epithelial cells (AECs) isolated from WT mice were treated with LPS (1μg/ml), live P. aeruginosa or heat-killed P. aeruginosa at a multiplicity of infection (MOI) 10. (A, B) After 4 h and 24 h incubation, expression of IL-36α (left panel) and IL-36γ (right panel) mRNA in PMs (A) and AECs (B) were analyzed by real-time PCR. (C, D) After 24 h incubation, PMs and AECs were treated with or without ATP (5mM) during 20 min of incubation, and then culture medium (CM) were harvested. The protein levels of IL-36α (left panel) and IL-36γ (right panel) in CM of treated PMs (C) and AECs (D) were measured by ELISA. Data (means ± SEM) are representative of two independent experiments. N.D.; not detected. * p<0.05, # p<0.01, § p<0.001, ¶ p<0.0001, N.S.; not significant, compared with medium only or as indicated. LPS; lipopolysaccharide, PA; Pseudomonas aeruginosa, HK PA; heat killed Pseudomonas aeruginosa, ATP; adenosine triphosphate.