Advanced Glycation End Products Promote Differentiation of CD4~＋ T Helper Cells toward Pro-inflammatory Response

• Published in: Journal of Huazhong University of Science and Technology. Medical sciences Vol. 34; no. 1; pp. 10 - 17
• Main Author: 韩晓群 龚作炯 徐三清 李汛 王立坤 伍仕敏 吴建红 杨华芬
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.02.2014; Department of Infectious Diseases, Renmin Hospital of Wuhan University, Wuhan 430060, China%Department of Pediatrics, Tongji Hospital Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030,China%Department of Infectious Diseases, Wuhan Medical Treatment Center, Wuhan 430023, China
• Subjects: Adult; Animals; Blotting, Western; Cattle; CD4; CD4-Positive T-Lymphocytes - drug effects; CD4-Positive T-Lymphocytes - metabolism; Cell Differentiation - drug effects; Cells, Cultured; Glucose - pharmacology; Glycation End Products, Advanced - pharmacology; HEK293 Cells; Humans; Interferon-gamma - metabolism; Interleukin-17 - metabolism; Medicine; Medicine & Public Health; mRNA表达; PPAR gamma - agonists; PPAR gamma - genetics; PPAR gamma - metabolism; PPARs; Prostaglandin D2 - analogs & derivatives; Prostaglandin D2 - pharmacology; Receptor for Advanced Glycation End Products; Receptors, Immunologic - genetics; Receptors, Immunologic - metabolism; Reverse Transcriptase Polymerase Chain Reaction; RNA Interference; Serum Albumin, Bovine - pharmacology; T-Lymphocytes, Regulatory - drug effects; T-Lymphocytes, Regulatory - metabolism; Th1 Cells - drug effects; Th1 Cells - metabolism; Th17 Cells - drug effects; Th17 Cells - metabolism; Western印迹; 反应; 晚期糖基化终产物; 细胞分化; 过氧化物酶体增殖物激活受体; CD4; T cell subsets; diabetes; advanced glycation end products; pro-inflammatory response; CD4+ T cell subsets
• ISSN: 1672-0733; 1993-1352
• DOI: 10.1007/s11596-014-1224-1

This study investigated the effect of advanced glycation end products（AGEs） on differentiation of na ve CD4＋T cells and the role of the receptor of AGEs（RAGE） and peroxisome proliferator-activated receptors（PPARs） activity in the process in order to gain insight into the mechanism of immunological disorders in diabetes. AGEs were prepared by the reaction of bovine serum albumin（BSA） with glucose. Human na ve CD4＋T cells, enriched from blood of healthy adult volunteers with negative selection assay, were cultured in vitro and treated with various agents including AGEs, BSA, high glucose, PGJ2 and PD68235 for indicated time. In short hairpin（sh） RNA knock-down experiment, na ve CD4＋T cells were transduced with media containing shRNA-lentivirus generated from lentiviral packaging cell line, Lent-XTM293 T cells. Surface and intracellular cytokine stainings were used for examination of CD4＋T cell phenotypes, and real-time PCR and Western blotting for detection of transcription factor mRNA and protein expression, respectively. The suppressive function of regulatory T（Treg） cells was determined by a [3H]-thymidine incorporation assay. The results showed that AGEs induced higher pro-inflammatory Th1/Th17 cells differentiated from na ve CD4＋T cells than the controls, whereas did not affect anti-inflammatory Treg cells. However, AGEs eliminated suppressive function of Treg cells. In addition, AGEs increased RAGE mRNA expression in na ve CD4＋T cells, and RAGE knock-down by shRNA eliminated the effect of AGEs on the differentiation of CD4＋T cells and the reduction of suppressive function of Treg cells. Furthermore, AGEs inhibited the mRNA expression of PPARγ, not PPARα; PPARγ agonist, PGJ2, inhibited the effect of AGEs on na ve CD4＋T cell differentiation and reversed the AGE-reduced suppressive function of Treg cells; on the other hand, PPARγ antagonist, PD68235, attenuated the blocking effect of RAGE shRNA on the role of AGEs. It was concluded that AGEs may promote CD4＋T cells development toward pro-inflammatory state, which is associated with increased RAGE mRNA expression and reduced PPARγ activity. ＋