Thyrotropin and CD40L Stimulate Interleukin-12 Expression in Fibrocytes: Implications for Pathogenesis of Thyroid-Associated Ophthalmopathy.
Increased numbers of bone marrow-derived progenitor cells, known as fibrocytes, populate the peripheral circulation, orbit, and thyroid of patients with Graves' disease (GD). These cells have been implicated in the development of thyroid-associated ophthalmopathy. They can differentiate into myofibroblasts or adipocytes, produce inflammatory cytokines, and remodel tissue. This study sought to determine whether thyrotropin (TSH) and CD40 ligand (CD40L), implicated in the pathogenesis of GD, induce interleukin-12 (IL-12) in human fibrocytes.



IL-12 protein concentrations and mRNA levels were measured by Luminex and real-time polymerase chain reaction, respectively. Flow cytometry assessed intracellular IL-12 concentrations. Vector containing IL-12p40 promoter was transfected into cultured fibrocytes, and promoter activity was monitored using luciferase assay.



TSH and CD40L stimulated intracellular IL-12 protein accumulation in peripheral blood fibrocytes. Inhibiting Akt and nuclear factor-κB (NF-κB) activity diminished IL-12 expression in fibrocytes, while TSH did not induce promoter activity. TSH-mediated IL-12 production required de novo synthesized proteins and augmented IL-12 mRNA stability. IL-12 production mediated by CD40L required tumor necrosis factor receptor-associated factor 6.



TSH and CD40L induce IL-12 expression in fibrocytes, and Akt and NF-κB mediate this activity. Given the importance of IL-12 in immune function, its production by fibrocytes may promote an inflammatory immune response and tissue remodeling in thyroid-associated ophthalmopathy.
1 Department of Ophthalmology and Visual Sciences, University of Michigan Medical School , Ann Arbor, Michigan.|2 Department of Ophthalmology, Tianjin Medical University Eye Hospital , Tianjin, People's Republic of China .
Tong
T
Wu
1 Department of Ophthalmology and Visual Sciences, University of Michigan Medical School , Ann Arbor, Michigan.
Tünde
T
Mester
1 Department of Ophthalmology and Visual Sciences, University of Michigan Medical School , Ann Arbor, Michigan.
Shivani
S
Gupta
2 Department of Ophthalmology, Tianjin Medical University Eye Hospital , Tianjin, People's Republic of China .
Fengyuan
F
Sun
1 Department of Ophthalmology and Visual Sciences, University of Michigan Medical School , Ann Arbor, Michigan.|3 Division of Metabolism, Endocrinology, and Diabetes, Department of Internal Medicine, University of Michigan Medical School , Ann Arbor, Michigan.
Terry J
TJ
Smith
1 Department of Ophthalmology and Visual Sciences, University of Michigan Medical School , Ann Arbor, Michigan.|4 Ann Arbor Veterans Administration Medical Center , Ann Arbor, Michigan.
Raymond S
RS
Douglas
D023201:CD40 Ligand / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism; D005434:Flow Cytometry; D006111:Graves Disease / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D018664:Interleukin-12 / Q000235:genetics / Q000378:metabolism*; D016328:NF-kappa B / Q000378:metabolism; D011401:Promoter Regions, Genetic; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013972:Thyrotropin / Q000494:pharmacology*
D016328:NF-kappa B; D023201:CD40 Ligand; D018664:Interleukin-12; D013972:Thyrotropin; D051057:Proto-Oncogene Proteins c-akt
Graves' disease; autoimmunity; fibrocyte; interleukin-12; thyroid-associated ophthalmopathy