title: Increased Toll-Like Receptors Activity and TLR Ligands in Patients with Autoimmune Thyroid Diseases.
abstract: Autoimmune thyroid disease (AITD) is an organ-specific disorder due to the interplay between environmental and genetic factors. Toll-like receptors (TLRs) are pattern recognition receptors expressed abundantly on monocytes. There is a paucity of data on TLR expression in AITD. The aim of this study was to examine TLR expression, activation, ligands, and downstream signaling adaptors in peripheral blood mononuclear cells (PBMCs) extracted from untreated AITD patients and healthy controls.
  
  
  
  We isolated PBMC of 30 healthy controls, 36 patients with untreated Hashimoto's thyroiditis, and 30 patients with newly onset Graves' disease. TLR mRNA, protein expression, TLR ligands, and TLR adaptor molecules were measured using real-time PCR, Western blot, flow cytometry, and enzyme-linked immunosorbent assay (ELISA). PBMC was simulated with TLR agonists. The effects of TLR agonists on the viability of human PBMC were evaluated using the MTT assay. The supernatants of cell cultures were measured for the pro-inflammatory cytokines, interleukin (IL)-6, tumor necrosis factor alpha (TNF-α), and IL-10 by ELISA.
  
  
  
  TLR2, TLR3, TLR9, and TLR10 mRNA were significantly increased in AITD patients compared with controls. TLR2, TLR3, TLR9, high mobility group box 1 (HMGB1), and RAGE expression on monocytes was higher in patients than control at baseline and TLR agonists' stimulation. The release of TNF-α and IL-6 was significantly increased in PBMCs from AITD patients with TLR agonists, while IL-10 was significantly decreased. Downstream targets of TLR, myeloid differentiation factor 88 (MyD88), and myeloid toll/IL-1 receptor-domain containing adaptor-inducing interferon-β were significantly elevated in AITD patients. Levels of TLR2 ligands, HMGB1, and heat shock protein 60 were significantly elevated in AITD patients compared with those in controls and positively correlated with TgAb and TPOAb, while sRAGE concentration was significantly decreased in AITD patients.
  
  
  
  This work is the first to show that TLR2, TLR3, and TLR9 expression and activation are elevated in the PBMCs of patients with AITD and TLRs may participate in the pathogenesis of AITD.
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China
authors.forename: Shiqiao
authors.initials: S
authors.lastname: Peng
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China|Department of Endocrinology and Metabolism, The First Hospital of China Medical University, Shenyang, China
authors.forename: Chenyan
authors.initials: C
authors.lastname: Li
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China|Department of Laboratory Medicine, The First Hospital of China Medical University, Shenyang, China
authors.forename: Xinyi
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China|Department of Intensive Care Unit, Affiliated Hospital of Qingdao University, Qingdao, China
authors.forename: Xin
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China
authors.forename: Cheng
authors.initials: C
authors.lastname: Han
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China|Department of Endocrinology, Sir Run Run Shaw Hospital, Affiliated to School of Medicine, Zhejiang University, Hangzhou, China
authors.forename: Ting
authors.initials: T
authors.lastname: Jin
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China|Department of Emergency, People’s Liberation Army No.202 Hospital, Shenyang, China
authors.forename: Shanshan
authors.initials: S
authors.lastname: Liu
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China
authors.forename: Xiaowen
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China
authors.forename: Hanyi
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China
authors.forename: Xue
authors.initials: X
authors.lastname: He
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China
authors.forename: Xiaochen
authors.initials: X
authors.lastname: Xie
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China|Department of Endocrinology and Metabolism, The First Hospital of China Medical University, Shenyang, China
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Yu
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China|Department of Endocrinology and Metabolism, The First Hospital of China Medical University, Shenyang, China
authors.forename: Chuyuan
authors.initials: C
authors.lastname: Wang
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China
authors.forename: Ling
authors.initials: L
authors.lastname: Shan
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China
authors.forename: Chenling
authors.initials: C
authors.lastname: Fan
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China|Department of Endocrinology and Metabolism, The First Hospital of China Medical University, Shenyang, China
authors.forename: Zhongyan
authors.initials: Z
authors.lastname: Shan
authors.affiliation: Department of Endocrinology and Metabolism, Institute of Endocrinology, Liaoning Provincial Key Laboratory of Endocrine Diseases, The First Affiliated Hospital of China Medical University, China Medical University, Shenyang, Liaoning, China|Department of Endocrinology and Metabolism, The First Hospital of China Medical University, Shenyang, China
authors.forename: Weiping
authors.initials: W
authors.lastname: Teng
keywords: autoimmune thyroid disease; innate immunity; pathogenesis; signaling; toll-like receptor