CD34− Orbital Fibroblasts From Patients With Thyroid-Associated Ophthalmopathy Modulate TNF-α Expression in CD34+ Fibroblasts and Fibrocytes
| Autor: | Tünde Mester, Priscila Novaes, Ana Beatriz Diniz Grisolia, Pei Mou, Terry J. Smith, Roshini Fernando, Aaron Trierweiler, Yan Lu, Stephen J. Atkins |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
endocrine system
Blotting Western Thyrotropin 030209 endocrinology & metabolism Antigens CD34 Enzyme-Linked Immunosorbent Assay orbital fibroblasts Antibodies Monoclonal Humanized Real-Time Polymerase Chain Reaction p38 Mitogen-Activated Protein Kinases Dexamethasone Flow cytometry 03 medical and health sciences 0302 clinical medicine Western blot medicine Humans RNA Messenger Receptor Glucocorticoids Cells Cultured Immunology and Microbiology Reporter gene Messenger RNA medicine.diagnostic_test Chemistry Tumor Necrosis Factor-alpha thyroid-associated ophthalmopathy fibrocytes Antibodies Monoclonal Fibroblasts Flow Cytometry Molecular biology 3. Good health Blot Graves Ophthalmopathy Real-time polymerase chain reaction Gene Expression Regulation TNF-α 030221 ophthalmology & optometry Tumor necrosis factor alpha Orbit |
| Zdroj: | Investigative Ophthalmology & Visual Science |
| ISSN: | 1552-5783 0146-0404 |
| Popis: | Purpose Orbital fibroblasts from patients with Graves' disease (GD-OF) express many different cytokines when treated with bovine thyrotropin (bTSH). The present study aimed to determine why TNF-α cannot be induced by bTSH in GD-OF. Methods Fibrocytes and GD-OFs were cultivated from donors who were patients in a busy academic medical center practice. Real-time PCR, Western blot analysis, reporter gene assays, cell transfections, mRNA stability assays, ELISA, and flow cytometry were performed. Results We found that bTSH induces TNF-α dramatically in fibrocytes but is undetectable in GD-OF. The induction in fibrocytes is a consequence of increased TNF-α gene promoter activity and is independent of ongoing protein synthesis. It could be attenuated by dexamethasone and the IGF-1 receptor inhibiting antibody, teprotumumab. When separated into pure CD34+ OF and CD34- OF subsets, TNF-α mRNA became highly inducible by bTSH in CD34+ OF but remained undetectable in CD34- OF. Conditioned medium from CD34- OF inhibited induction of TNF-α in fibrocytes. Conclusions Our data indicate that CD34- OF appear to release a soluble(s) factor that downregulates expression and induction by bTSH of TNF-α in fibrocytes and their derivative CD34+ OF. We proffer that CD34- OF produce an unidentified modulatory factor that attenuates TNF-α expression in GD-OF and may do so in the TAO orbit. |
| Databáze: | OpenAIRE |
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