Cigarette smoke extract induces thymic stromal lymphopoietin expression, leading to T_H2-type immune responses and airway inflammation

CSE exposure induces TSLP expression in the mouse lung. A-C, BALB/c mice were intranasally challenged with CSE or PBS every day for a total of 7 days (days 0-6). The mouse lungs were removed on day 7 (3 hours after the final challenge), and real-time PCR (Fig 1, A), Western blot analysis (n = 3 in each group; Fig 1, B), and immunohistochemical staining (Fig 1, C) for TSLP were performed. Quantitative analysis of the TSLP immunoreactivity is shown in Fig 1, D. Representative results of 3 independent experiments are shown. ^∗P < .05 in comparison with the corresponding control. GAPDH, Glyceraldehyde-3-phosphate dehydrogenase.

CSE-induced TSLP expression in the mouse lung depends on oxidative stress and TNF-α receptor I (TNFRI). A-C, BALB/c mice were intranasally challenged with CSE or PBS every day for a total of 7 days (days 0-6) with or without treatment with 150 mg/kg NAC. The mouse lungs were removed on day 7 (3 hours after the final challenge), and real-time PCR for TSLP (Fig 2, A), TNF-α (Fig 2, B), heme oxygenase 1 (HO-1; Fig 2, C), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was performed. D and E, BALB/c mice (Fig 2, D) or TNFRI-null mice and control C57BL/6 mice (Fig 2, E) were intratracheally challenged with CSE, and the mouse lungs were removed at the indicated times (Fig 2, D) or 3 hours after the challenge (Fig 2, E), and real-time RT-PCR was performed for TSLP and GAPDH. Representative results of 3 independent experiments are shown. ^∗P < .05 in comparison with the corresponding control.

Intranasal exposure of CSE simultaneously with OVA induces airway inflammation in mice. BALB/c mice were intranasally challenged with CSE, OVA, or both (or PBS) 5 times per week for 8 weeks with or without treatment with 15 mg/kg per mouse anti-TSLP antibody or control antibody every week. The mouse lungs were removed at 2 (A), 4 (B), and 8 (C) weeks during the CSE challenge, the OVA challenge, or both, and then the tissue sections were stained with hematoxylin and eosin. Representative results of 3 independent experiments are shown.

Intranasal exposure of CSE simultaneously with OVA induces OVA-specific T_H2-type immune responses in mice. BALB/c mice were intranasally challenged with CSE, OVA, or both (or PBS) 5 times per week for 8 weeks with or without treatment with 15 mg/kg per mouse anti-TSLP antibody or control antibody every week. The mice were killed at 3 hours after the final CSE challenge, OVA challenge, or both for further analysis. A-C, RNA was extracted from the mouse lungs, and real-time RT-PCR was performed for mEAR2 (Fig 4, A), GATA-3 (Fig 4, B), IL-13 (Fig 4, C), and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). D, The mouse sera were collected, and the concentrations of OVA-specific IgE antibody were measured by means of ELISA. E and F, Spleen cells were collected and stimulated with 100 μg/mL OVA for 72 hours in vitro. The concentrations of IL-4 (Fig 4, E) and IL-13 (Fig 4, F) in the culture supernatants were measured by means of ELISA. Representative results of 3 independent experiments are shown. ^∗P < .05 in comparison with the corresponding control.