Ming-Chi Lu1, Chia-Li Yu2, Hua-Chien Chen2, Hui-Chun Yu2, Hsien-Bin Huang2, Ning-Sheng Lai3. 1. Division of Allergy, Immunology and Rheumatology, Dalin Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Chiayi, School of Medicine, Tzu Chi University, Hualien, Department of Internal Medicine, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Department of Biomedical Sciences, Chang Gung University, Taoyuan and Department of Life Science and Institute of Molecular Biology, National Chung Cheng University, Chiayi, Taiwan. Division of Allergy, Immunology and Rheumatology, Dalin Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Chiayi, School of Medicine, Tzu Chi University, Hualien, Department of Internal Medicine, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Department of Biomedical Sciences, Chang Gung University, Taoyuan and Department of Life Science and Institute of Molecular Biology, National Chung Cheng University, Chiayi, Taiwan. 2. Division of Allergy, Immunology and Rheumatology, Dalin Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Chiayi, School of Medicine, Tzu Chi University, Hualien, Department of Internal Medicine, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Department of Biomedical Sciences, Chang Gung University, Taoyuan and Department of Life Science and Institute of Molecular Biology, National Chung Cheng University, Chiayi, Taiwan. 3. Division of Allergy, Immunology and Rheumatology, Dalin Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Chiayi, School of Medicine, Tzu Chi University, Hualien, Department of Internal Medicine, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Department of Biomedical Sciences, Chang Gung University, Taoyuan and Department of Life Science and Institute of Molecular Biology, National Chung Cheng University, Chiayi, Taiwan. Division of Allergy, Immunology and Rheumatology, Dalin Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Chiayi, School of Medicine, Tzu Chi University, Hualien, Department of Internal Medicine, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Department of Biomedical Sciences, Chang Gung University, Taoyuan and Department of Life Science and Institute of Molecular Biology, National Chung Cheng University, Chiayi, Taiwan. tzuchilai@gmail.com.
Abstract
OBJECTIVE: The aim of this study was to investigate the pathogenic role of calcium (Ca(2+)) influx-regulated microRNAs (miRNAs) in T cells from patients with SLE. METHODS: Expression profiles of 270 human miRNAs in Jurkat cells co-cultured with or without ionomycin were analysed by real-time PCR. Differential expression of miRNAs in T cell samples from 28 patients with SLE (SLE T cells) and 20 healthy controls were investigated using western blot analysis of proteins expressed by respective miRNA target transcripts. Transfection studies were conducted to investigate miRNA-specific biological functions. RESULTS: Initial analysis revealed differential expression of nine miRNAs in Jurkat cells after co-culture with ionomycin. Of these, miR-524-5p and miR-449b were overexpressed in SLE T cells. Levels of expressed miR-524-5p showed a significant direct correlation with the SLEDAI. Transfection of Jurkat cells with miR-524-5p mimic suppressed Jagged-1 and Hes-1 protein expression. Likewise, expression of both Jagged-1 and Hes-1 proteins were diminished in SLE T cells. Upon activation of Jurkat cells transfected with miR-524-5p mimic, production of IFN-γ increased but the apoptotic rate was unaffected. CONCLUSION: In SLE T cells, miR-524-5p and miR-449b (both regulated by Ca(2+) influx) were overexpressed. Moreover, increased miR-524-5p expression, as shown by patients with SLE, directly paralleled disease activity (SLEDAI). Transfection of miR-524-5p also enhanced IFN-γ production in activated Jurkat cells.
OBJECTIVE: The aim of this study was to investigate the pathogenic role of calcium (Ca(2+)) influx-regulated microRNAs (miRNAs) in T cells from patients with SLE. METHODS: Expression profiles of 270 human miRNAs in Jurkat cells co-cultured with or without ionomycin were analysed by real-time PCR. Differential expression of miRNAs in T cell samples from 28 patients with SLE (SLE T cells) and 20 healthy controls were investigated using western blot analysis of proteins expressed by respective miRNA target transcripts. Transfection studies were conducted to investigate miRNA-specific biological functions. RESULTS: Initial analysis revealed differential expression of nine miRNAs in Jurkat cells after co-culture with ionomycin. Of these, miR-524-5p and miR-449b were overexpressed in SLE T cells. Levels of expressed miR-524-5p showed a significant direct correlation with the SLEDAI. Transfection of Jurkat cells with miR-524-5p mimic suppressed Jagged-1 and Hes-1 protein expression. Likewise, expression of both Jagged-1 and Hes-1 proteins were diminished in SLE T cells. Upon activation of Jurkat cells transfected with miR-524-5p mimic, production of IFN-γ increased but the apoptotic rate was unaffected. CONCLUSION: In SLE T cells, miR-524-5p and miR-449b (both regulated by Ca(2+) influx) were overexpressed. Moreover, increased miR-524-5p expression, as shown by patients with SLE, directly paralleled disease activity (SLEDAI). Transfection of miR-524-5p also enhanced IFN-γ production in activated Jurkat cells.
Authors: Elizabeth J Brant; Edward A Rietman; Giannoula Lakka Klement; Marco Cavaglia; Jack A Tuszynski Journal: PLoS One Date: 2020-03-19 Impact factor: 3.240