Cytokine regulation of IL-13Rα2 and IL-13Rα1 in vivo and in vitro

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Cytokine regulation of IL-13Rα2 and IL-13Rα1 in vivo and in vitro Tao Zheng, MDa, Zhou Zhu, MD, PhDa, Wei Liu, MDa, Chun Geun Lee, MD, PhDa, Qingsheng Chen, MDa, Robert J. Homer, MD, PhDb,c, Jack A. Elias, MDa Journal of Allergy and Clinical Immunology Volume 111, Issue 4, Pages 720-728 (April 2003) DOI: 10.1067/mai.2003.1383 Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 1 Effects of IL-13 on IL-13Rα2 and IL-13Rα1 mRNA accumulation in vivo. A , One-month-old CC10–rtTA–tTS–IL-13 mice were randomized to receive normal water or dox water for 4 weeks, and Northern blot analysis was used to quantitate IL-13Rα2, IL-13Rα1, and β-actin mRNA, as noted. B, Densitometric evaluations were undertaken of 5 replicate experiments (*P < .01 vs transgene(–) controls; Student t test). C, Whole lung RNA was obtained from CC10–IL-13 transgenic mice and transgene(–) littermate controls at 2 months of age, and RT-PCR was used to evaluate the levels of mRNA encoding IL-13Rα2, IL-13Rα1, and β-actin, as noted. Journal of Allergy and Clinical Immunology 2003 111, 720-728DOI: (10.1067/mai.2003.1383) Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 2 Localization of IL-13Rα2 mRNA accumulation in vivo. In each panel, ISH was used to compare the levels of IL-13Rα2 mRNA in CC10–IL-13 transgene(+) and transgene(–) (wild type [WT] ) mice. Sense and antisense probes were used as described in the Methods section. Staining in macrophages (indicated by arrows in A ), small airway epithelial cells (B ), and large airway epithelial cells (C ) can be seen. Journal of Allergy and Clinical Immunology 2003 111, 720-728DOI: (10.1067/mai.2003.1383) Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 3 Localization of IL-13Rα1 in vivo. ISH was used to compare the levels of mRNA encoding IL-13Rα1 through use of sense and antisense probes. Epithelial staining (large arrows) and macrophage staining (small arrows) are highlighted. Journal of Allergy and Clinical Immunology 2003 111, 720-728DOI: (10.1067/mai.2003.1383) Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 4 Effects of IL-4, IL-10, and IFN-γ on IL-13Rα2 and IL-13Rα1 mRNA accumulation. These are represent-ative experiments of N = 4. The levels of mRNA encoding IL-13Rα2 and IL-13Rα1 were compared in lungs from 2-month-old CC10–IL-4 transgene(+) and transgene(–) mice (A ) and 2- and 3-month (m) –old CC10–IL-10 transgene(+) and transgene(–) mice (B ). Also illustrated are the levels of mRNA encoding IL-13Rα2 and IL-13Rα1 in lungs from CC10–rtTA–IFN-γ mice that have been randomized to receive normal water or dox water at 1 month of age and were maintained on this regimen for 4 weeks (C ). Journal of Allergy and Clinical Immunology 2003 111, 720-728DOI: (10.1067/mai.2003.1383) Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 5 Role of IL-4Rα and blood leukocytes in IL-13 regulation of IL-13Rα2 and IL-13Rα1. A, The levels of mRNA encoding IL-13Rα2 and IL-13Rα1 in lungs from 2-month old transgene(+) CC10–IL-13 mice that had (+/+) or (–/–) (null mutant) IL-4Rα loci were evaluated by means of RT-PCR. B, Lung organ cultures were established from lungs of wild-type mice and incubated in the presence and in the absence of rIL-13, as noted. The levels of mRNA encoding IL-13Rα2 and IL-13Rα1 were evaluated by means of RT-PCR. Journal of Allergy and Clinical Immunology 2003 111, 720-728DOI: (10.1067/mai.2003.1383) Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 6 Effects of IL-13, IFN-γ, and IL-4 on the levels of mRNA encoding IL-13Rα2 and IL-13Rα1 in NHBE cells and RAW cells. A through C, Representative experiments (N = 3) defining the kinetics of the effects of human (h) cytokines on NHBE cells. D through F, Representative experiments (N = 3) defining the effects of murine (m) cytokines on RAW cells. Journal of Allergy and Clinical Immunology 2003 111, 720-728DOI: (10.1067/mai.2003.1383) Copyright © 2003 Mosby, Inc. Terms and Conditions

Fig. 7 Effects of IL-13 on IL-13Rα2 protein production. These are representative experiments of N = 3. A, Mononuclear cells were isolated from lungs from wild-type mice and incubated with 35S-translabel in the presence (+) and in the absence (–) of rmIL-13 (50 ng/mL). B, NHBE cells were incubated with 35S-translabel in the presence and in the absence of rhIL-13 (50 ng/mL) for 24 hours. The amount of IL-13Rα2 protein that was produced during these intervals was evaluated by means of immunoprecipitation, as described in the Methods section. Journal of Allergy and Clinical Immunology 2003 111, 720-728DOI: (10.1067/mai.2003.1383) Copyright © 2003 Mosby, Inc. Terms and Conditions