Institute of Allergy and Clinical Immunology, Seoul National University Medical Research Center, Seoul, Republic of Korea.
Division of Allergy and Clinical Immunology, Department of Asan Medical Center, University of Ulsan College of Medicine, Seoul, Republic of Korea.
Department of Internal Medicine, Kyung Hee University Medical Center, Seoul, Republic of Korea.
Department of Internal Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea.
Institute of Allergy and Clinical Immunology, Seoul National University Medical Research Center, Seoul, Republic of Korea. guinea71@snu.ac.kr.
Department of Internal Medicine, Seoul National University College of Medicine, Seoul, Republic of Korea. guinea71@snu.ac.kr.
Abstract
Nonallergic eosinophilic asthma (NAEA) is a clinically distinct subtype of asthma. Thus far, the pathophysiologic mechanisms underlying NAEA have not been fully elucidated. This study aimed to determine the role of IL-23 in the pathogenesis of NAEA. We developed a murine model of NAEA using recombinant IL-23 (rIL-23) plus a nonspecific airway irritant [polyinosinic-polycytidylic acid (polyI:C) or diesel exhaust particles (DEPs)] and investigated whether IL-23 plays an important role in the development of NAEA. Intranasal administration of rIL-23 (0.1 μg/mouse) plus polyI:C (0.01 μg/mouse) or DEPs (10 μg/mouse) without allergen resulted in methacholine bronchial hyperresponsiveness and eosinophilic airway inflammation in mice, which are characteristic features of NAEA. rIL-23 plus a low dose nonspecific airway irritants induced the release of innate cytokines from airway epithelium, including IL-33, thymic stromal lymphopoietin and IL-1β; these factors activated types 2 and 3 innate lymphoid cells (ILC2s and ILC3s). ILC2s and ILC3s, but not CD4+ T cells (i.e., adaptive immune cells), were important in the development of NAEA. In addition, we observed that IL-23 receptor expressions increased in airway epithelial cells, which suggests the existence of a positive autocrine loop in our murine model of NAEA. To our knowledge, this is the first report in which administration of rIL-23 plus a nonspecific airway irritant (polyI:C or DEPs) without allergen resulted in features of NAEA in mice similar to those found in humans. IL-23 may constitute a therapeutic target for NAEA in humans.
Figures
Fig. 1 Determination of recombinant interleukin-23 (rIL-23) dose for stimulation. a Experimental protocol. b Airway resistance at doses of 50 and 100 mg/mL methacholine (MCh). c Inflammatory cells in bronchoalveolar lavage fluid (eosinophils are magnified). d IL-13-producing or IL-5-producing type 2 innate lymphoid cells (CD45+Lin− ICOS+) in lung homogenate. e IL-17A-producing type 3 innate lymphoid cells (CD45+Lin− CD25+) in lung homogenate. f, g IL-13-producing or IL-17A-producing CD4+ T cells in lung homogenate. h IL-33 and TSLP levels in lung homogenate. *P < 0.05, **P < 0.01 between two groups; #P < 0.05, ##P < 0.01 compared to the control group; AHR airway hyperresponsiveness; PBS phosphate-buffered saline; Macro macrophage; Neutro neutrophil; Eosino eosinophil; Lympho lymphocyte.Fig. 2 Nonallergic eosinophilic asthma phenotype of the polyI:C/rIL-23 model. a Airway resistance at doses of 50 and 100 mg/mL methacholine (MCh). b Eosinophils, neutrophils, and lymphocytes in bronchoalveolar lavage fluid. c CD11b+siglecF+CD11c− cells in bronchoalveolar lavage fluid. d IL-13-producing or IL-5-producing type 2 innate lymphoid cells (CD45+Lin− ICOS+) in lung homogenate. e IL-17A-producing type 3 innate lymphoid cells (CD45+Lin− CD25+) in lung homogenate. f, g IL-13-producing or IL-17A-producing CD4+ T cells in lung homogenate. h IL-33, TSLP, and IL-1β levels in lung homogenate. *P < 0.05, **P < 0.01 between two groups; #P < 0.05 compared to the control group; ns, not significant; PBS phosphate-buffered saline.Fig. 3 Nonallergic eosinophilic asthma phenotype of the DEP/rIL-23 model. a Airway resistance at doses of 50 and 100 mg/mL methacholine (MCh). b Eosinophils, neutrophils, and lymphocytes in bronchoalveolar lavage fluid. c CD11b+siglecF+CD11c− cells in bronchoalveolar lavage fluid. d IL-13 or IL5-producing type 2 innate lymphoid cells (CD45+Lin− ICOS+) in lung homogenate. e IL-17A-producing type 3 innate lymphoid cells (CD45+Lin− CD25+) in lung homogenate. f, g IL-13-producing or IL-17A-producing CD4+ T cells in lung homogenate. h IL-33, TSLP, and IL-1β levels in lung homogenate. *P< 0.05, **P< 0.01 between two groups; #P< 0.05 compared to the control group; DEP diesel exhaust particle; ns, not significant; PBS phosphate-buffered saline.Fig. 4 Immunohistochemistry findings. a IL-33. b Comparison of area% of IL-33. c TSLP. d Comparison of area% of TSLP. e α-SMA. f Comparison of area% of α-SMA. g IL-23R. h Comparison of area% of IL-23R. Magnification, ×40. Area% was quantified using ImageJ software. *P < 0.05 between two groups. DEP diesel exhaust particle; α-SMA alpha-smooth muscle actin; PBS phosphate-buffered saline.Fig. 5 Effect of polyI:C, DEP, and rIL-23 treatments on MLE12 cells. a IL-23 production by MLE12 cells treated with different doses of polyI:C or DEP. b IL-33 production by MLE12 cells treated with different doses of polyI:C or DEPs, with or without anti-IL-23 antibodiesy. c TSLP production by MLE12 cells treated with different doses of polyI:C or DEPs, with or without anti-IL-23 antibodies. d IL-33 production by MLE12 cells treated with different doses of rIL-23. e TSLP production by MLE12 cells treated with different doses of rIL-23. *P < 0.05 between two groups; #P < 0.05, ##P < 0.01 compared to medium only; DEP diesel exhaust particle.Fig. 6 Effect of exposure to polyI:C/rIL-23-treated or DEP/rIL-23-treated MLE12 cells on ILCs. a IL-33 levels in the nuclei of MLE12 cells. b TSLP levels in the cytosol of MLE12 cells. c IL-13 levels in the supernatant of ILC2s cocultured with MLE12 cells treated with polyI:C or DEPs, with or without rIL-23. d IL-5 levels in the supernatant of ILC2s cocultured with MLE12 cells treated with polyI:C or DEPs, with or without rIL-23. *P < 0.05, **P < 0.01 between two groups; DEP diesel exhaust particle.Fig. 7 Effects of IL-23R overexpression in MLE12 cells treated with polyI:C/rIL-23 or DEP/rIL-23. a IL-33 levels in the nuclei of MLE12 cells. b TSLP levels in the cytosol of MLE12 cells. *P < 0.05 between two groups; #P< 0.05 compared to medium only; DEP diesel exhaust particle.Fig. 8 A plausible mechanism for the development of nonallergic eosinophilic asthma. a Airway epithelial cells stimulated by high doses of IL-23. High-dose IL-23 binds to IL-23R and induces the secretion of IL-33, TSLP, and IL-1β from airway epithelial cells; these factors can activate ILC2s and ILC3s. Activated ILC2s and ILC3s cause eosinophilic airway inflammation and AHR. In addition, high-dose IL-23 contributes to the development of AHR by directly activating Th17 cells and neutrophils. b Airway epithelial cells stimulated by low doses of IL-23 plus nonspecific airway irritants (polyI:C or DEPs). Low-dose IL-23 plus nonspecific airway irritants induce the secretion of a small amount of IL-23 from airway epithelial cells and increases the expression of IL-23R on airway epithelial cells. The secreted IL-23 forms a positive autocrine loop by binding to the increased IL-23R, thereby inducing secretion of IL-33, TSLP, and IL-1β (as with the high-dose of IL-23). Activated ILC2s and ILC3s cause eosinophilic airway inflammation and AHR. ASMC airway smooth muscle cell; AHR airway hyperresponsiveness.Fig. 9IL-23A and IL-23R gene expression levels in induced sputum obtained from patients with nonallergic eosinophilic asthma, allergic eosinophilic asthma, and normal controls. a IL-23A. b IL-23R. **P < 0.01 between two groups; #P < 0.05 compared to normal controls; NAEA nonallergic eosinophilic asthma; AEA allergic eosinophilic asthma (details are provided in the supplementary information).
그림 출처: PubMed Central 공개 논문 (PMC7000690)
Conflict of interest statement
The authors declare that they have no conflict of interest.
그 밖의 정보
출판 형태
Journal Article, Research Support, Non-U.S. Gov't
저널
Experimental & molecular medicine · eISSN 2092-6413
