Chia-Wei Hsieh1,2, Yi-Ming Chen1,2, Chi-Chen Lin1,2, Kuo-Tung Tang1,2, Hsin-Hua Chen1,2, Wei-Ting Hung1,2, Kuo-Lung Lai1,2, Der-Yuan Chen3,4. 1. From the Division of Allergy, Immunology and Rheumatology, and the Department of Medical Education and Research, Taichung Veterans General Hospital, and PhD Program in Translational Medicine, and Institute of Biomedical Science and Rong Hsing Research Center for Translational Medicine, National Chung Hsing University, Taichung; Faculty of Medicine, National Yang Ming University, Taipei, Taiwan. 2. C.W. Hsieh, MD, Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, and PhD Program in Translational Medicine, National Chung Hsing University; Y.M. Chen, MD, PhD, Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, and Department of Medical Education and Research, Taichung Veterans General Hospital, and Faculty of Medicine, National Yang Ming University, and PhD Program in Translational Medicine, National Chung Hsing University; C.C. Lin, PhD, PhD Program in Translational Medicine, National Chung Hsing University, and Institute of Biomedical Science and Rong Hsing Research Center for Translational Medicine, National Chung Hsing University; K.T. Tang, MD, Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, and PhD Program in Translational Medicine, National Chung Hsing University; H.H. Chen, MD, PhD, Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, and Department of Medical Education and Research, Taichung Veterans General Hospital, and Faculty of Medicine, National Yang Ming University; W.T. Hung, MD, Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, and Department of Medical Education and Research, Taichung Veterans General Hospital; K.L. Lai, MD, Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, and Institute of Biomedical Science and Rong Hsing Research Center for Translational Medicine, National Chung Hsing University; D.Y. Chen, MD, PhD, Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, and Department of Medical Education and Research, Taichung Veterans General Hospital, and Faculty of Medicine, National Yang Ming University, and PhD Program in Translational Medicine, National Chung Hsing University, and Institute of Biomedical Science and Rong Hsing Research Center for Translational Medicine, National Chung Hsing University. Dr. Y.M. Chen and C.C. Lin contributed equally to this work. 3. From the Division of Allergy, Immunology and Rheumatology, and the Department of Medical Education and Research, Taichung Veterans General Hospital, and PhD Program in Translational Medicine, and Institute of Biomedical Science and Rong Hsing Research Center for Translational Medicine, National Chung Hsing University, Taichung; Faculty of Medicine, National Yang Ming University, Taipei, Taiwan. dychen@vghtc.gov.tw. 4. C.W. Hsieh, MD, Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, and PhD Program in Translational Medicine, National Chung Hsing University; Y.M. Chen, MD, PhD, Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, and Department of Medical Education and Research, Taichung Veterans General Hospital, and Faculty of Medicine, National Yang Ming University, and PhD Program in Translational Medicine, National Chung Hsing University; C.C. Lin, PhD, PhD Program in Translational Medicine, National Chung Hsing University, and Institute of Biomedical Science and Rong Hsing Research Center for Translational Medicine, National Chung Hsing University; K.T. Tang, MD, Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, and PhD Program in Translational Medicine, National Chung Hsing University; H.H. Chen, MD, PhD, Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, and Department of Medical Education and Research, Taichung Veterans General Hospital, and Faculty of Medicine, National Yang Ming University; W.T. Hung, MD, Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, and Department of Medical Education and Research, Taichung Veterans General Hospital; K.L. Lai, MD, Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, and Institute of Biomedical Science and Rong Hsing Research Center for Translational Medicine, National Chung Hsing University; D.Y. Chen, MD, PhD, Division of Allergy, Immunology and Rheumatology, Taichung Veterans General Hospital, and Department of Medical Education and Research, Taichung Veterans General Hospital, and Faculty of Medicine, National Yang Ming University, and PhD Program in Translational Medicine, National Chung Hsing University, and Institute of Biomedical Science and Rong Hsing Research Center for Translational Medicine, National Chung Hsing University. Dr. Y.M. Chen and C.C. Lin contributed equally to this work. dychen@vghtc.gov.tw.
Abstract
OBJECTIVE: The dysregulation of the NLRP3 (NLR containing a pyrin domain) inflammasome is involved in autoinflammatory diseases. Adult-onset Still disease (AOSD) is regarded as an autoinflammatory disease. However, the pathogenic involvement of NLRP3 inflammasome in AOSD remains unclear and NLRP3 activators in AOSD are currently unknown. METHODS: The mRNA expression of NLRP3 inflammasome signaling in peripheral blood mononuclear cells (PBMC) from 34 patients with AOSD and 14 healthy subjects was determined using quantitative-PCR (qPCR). The changes in mRNA and protein levels of NLRP3 inflammasome signaling in PBMC treated with the potential activator [imiquimod (IMQ)] or inhibitor of NLRP3 were evaluated using qPCR and immunoblotting, respectively. The supernatant levels of interleukin (IL)-1β and IL-18 were determined by ELISA. RESULTS: Significantly higher mRNA levels of NLRP3 inflammasome signaling were observed in patients with AOSD compared with healthy controls. NLRP3 expressions were positively correlated with disease activity in patients with AOSD. IMQ (an effective Toll-like receptor 7 ligand; 10 µg/ml and 25 µg/ml) stimulation of PBMC from patients with AOSD induced dose-dependent increases of mRNA expression of NLRP3 (mean ± standard error of the mean, 2.06 ± 0.46 and 6.05 ± 1.84, respectively), caspase-1 (1.81 ± 0.23 and 4.25 ± 0.48), IL-1β (5.68 ± 1.51 and 12.13 ± 3.71), and IL-18 (2.32 ± 0.37 and 4.81 ± 0.51) compared with controls (all p < 0.005). IMQ stimulation of PBMC from patients similarly induced greater increases in protein expressions of NLRP3 inflammasome compared with controls. The protein expressions of NLRP3, IL-1β, and IL-18 on PBMC significantly decreased after treatment with NLRP3 inhibitor in patients with AOSD. CONCLUSION: Increased expression of NLRP3 inflammasome and its positive correlation with disease activity in AOSD suggest its involvement in disease pathogenesis. IMQ upregulated expressions of NLRP3 inflammasome signaling, and IMQ might be an activator of NLRP3 inflammasome in AOSD.
OBJECTIVE: The dysregulation of the NLRP3 (NLR containing a pyrin domain) inflammasome is involved in autoinflammatory diseases. Adult-onset Still disease (AOSD) is regarded as an autoinflammatory disease. However, the pathogenic involvement of NLRP3 inflammasome in AOSD remains unclear and NLRP3 activators in AOSD are currently unknown. METHODS: The mRNA expression of NLRP3 inflammasome signaling in peripheral blood mononuclear cells (PBMC) from 34 patients with AOSD and 14 healthy subjects was determined using quantitative-PCR (qPCR). The changes in mRNA and protein levels of NLRP3 inflammasome signaling in PBMC treated with the potential activator [imiquimod (IMQ)] or inhibitor of NLRP3 were evaluated using qPCR and immunoblotting, respectively. The supernatant levels of interleukin (IL)-1β and IL-18 were determined by ELISA. RESULTS: Significantly higher mRNA levels of NLRP3 inflammasome signaling were observed in patients with AOSD compared with healthy controls. NLRP3 expressions were positively correlated with disease activity in patients with AOSD. IMQ (an effective Toll-like receptor 7 ligand; 10 µg/ml and 25 µg/ml) stimulation of PBMC from patients with AOSD induced dose-dependent increases of mRNA expression of NLRP3 (mean ± standard error of the mean, 2.06 ± 0.46 and 6.05 ± 1.84, respectively), caspase-1 (1.81 ± 0.23 and 4.25 ± 0.48), IL-1β (5.68 ± 1.51 and 12.13 ± 3.71), and IL-18 (2.32 ± 0.37 and 4.81 ± 0.51) compared with controls (all p < 0.005). IMQ stimulation of PBMC from patients similarly induced greater increases in protein expressions of NLRP3 inflammasome compared with controls. The protein expressions of NLRP3, IL-1β, and IL-18 on PBMC significantly decreased after treatment with NLRP3 inhibitor in patients with AOSD. CONCLUSION: Increased expression of NLRP3 inflammasome and its positive correlation with disease activity in AOSD suggest its involvement in disease pathogenesis. IMQ upregulated expressions of NLRP3 inflammasome signaling, and IMQ might be an activator of NLRP3 inflammasome in AOSD.
Entities:
Keywords:
ACTIVATOR; ADULT-ONSET STILL DISEASE; IMIQUIMOD; INFLAMMASOME; NLRP3