Synergistic effect of regulatory T cells and proinflammatory cytokines in angiogenesis in the endometriotic milieu

Autor: Xue-Zhen Luo, Da-Jin Li, Wen-Jie Zhou, Xiao-Qiu Wang, Yu Tao
Rok vydání: 2017
Předmět:
Adult
0301 basic medicine
Stromal cell
MAP Kinase Signaling System
Angiogenesis
Endometriosis
Enzyme-Linked Immunosorbent Assay
Cell Communication
T-Lymphocytes
Regulatory
Monocytes
Proinflammatory cytokine
Endometrium
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
Cell Movement
Human Umbilical Vein Endothelial Cells
Humans
Medicine
IL-2 receptor
Cells
Cultured
Chemokine CCL22
Immunosuppression Therapy
030219 obstetrics & reproductive medicine
Neovascularization
Pathologic
business.industry
Rehabilitation
Obstetrics and Gynecology
Interleukin
FOXP3
Coculture Techniques
Vascular endothelial growth factor
030104 developmental biology
Reproductive Medicine
chemistry
Cancer research
Female
Tumor necrosis factor alpha
Chemokine CCL17
Endothelium
Vascular
Inflammation Mediators
Stromal Cells
business
Biomarkers
Zdroj: Human Reproduction. 32:1304-1317
ISSN: 1460-2350
0268-1161
Popis: STUDY QUESTION Do regulatory T cells (Tregs) contribute to angiogenesis in endometriosis? SUMMARY ANSWER High levels of CCL17 and CCL22 cause the recruitment of Tregs, upregulate the immunosuppression of Tregs and, in turn, may promote angiogenesis in endometrial cells in synergy with proinflammatory cytokines. WHAT IS ALREADY KNOWN The peritoneal fluid of patients with endometriosis has a higher percentage of Tregs than that of normal individuals; however, the regulatory role of Tregs in the disease remains unclear. STUDY DESIGN, SIZE, DURATION This study used primary human endometrial stromal cells (ESCs), monocytes (Mo), Tregs and human umbilical vein endothelial cells (HUVECs). All experiments were performed at least three times. PARTICIPANTS/MATERIALS, SETTING, METHODS The migration of Tregs was evaluated by the transwell migration assay. The activation of extracellular signal-regulated kinase (ERK)1/2, c-Jun N-terminal kinase and p38 signaling pathways was examined using the In-Cell WesternTM (LI-COR®) western blot analysis system, as well as by traditional western blot analysis. Changes in the expression of CCL22, CCL17, transforming growth factor-beta 1 (TGF-β1), Interleukin (IL)-1β, tumor necrosis factor alpha (TNF-α), IL-8 and vascular endothelial growth factor (VEGF) in cell-culture supernatant were detected by ELISA. We analyzed the Tregs by multicolor flow cytometry to directly test the expression of CCR4, CD4, CD25, Foxp3, CTLA-4, CD39 and CD73. MAIN RESULTS AND THE ROLE OF CHANCE Our results showed that ESCs-Mo co-culture produced significantly higher levels of CCL22 and CCL17 than ESCs or Mo cultured alone, and that estradiol (E2) or progesterone (P) further promoted this upregulation, demonstrating stronger chemotaxis on Tregs. The co-culture of ESCs with Mo stimulated TGF-β1 secretion by Tregs, which could be inhibited by anti-CCL22 or/and anti-CCL17 neutralizing antibodies (Abs). The expression of CCR4 by Tregs was upregulated in ESCs-Mo co-culture, especially by treatment with E2 and/or P, and this effect could be abolished by anti-CCL22 and/or anti-CCL17-neutralizing Abs. The Treg-ESCs-Mo co-culture treated with E2 (10-8 mol/l) and P (10-8 mol/l) could enhance the immunosuppression of Tregs, as proved by the elevated expression of Foxp3, CTLA-4, CD39 and CD73 on Tregs. ESCs-Mo co-culture could significantly promote the secretion of IL-1β and TNF-α. TGF-β1 from Tregs could activate p38/ERK1/2 signaling pathways in ESCs, and IL-1β and TNF-α produced by ESCs-Mo co-culture had synergistic roles with TGF-β1. TGF-β1 and the proinflammatory cytokines IL-1β or TNF-α could synergistically promote IL-8 and VEGF expression in ESCs via the p38/ERK1/2 signaling pathways. The high levels of IL-8 and VEGF in the supernatant of ESCs stimulated the angiogenesis of HUVECs. LARGE SCALE DATA None. LIMITATIONS, REASONS FOR CAUTION This study was only performed in vitro using eutopic ESCs, instead of ectopic cells, from endometriosis patients. Therefore, it is necessary to do further experiments to determine whether Tregs promote angiogenesis in the endometriotic milieu in synergy with proinflammatory cytokines in vivo. WIDER IMPLICATIONS OF THE FINDINGS Co-targeting Tregs and proinflammatory cytokines may be an effective treatment for endometriosis. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by Ministry of Science and Technology of China 2015CB943300 to L.D.-J.; National Natural Science Foundation of China, item number 81200425 to W.X.-Q.; National Natural Science Foundation of China, item number 81471548 to L.D.-J.; and The Research Fund for the Doctoral Program of Higher Education of China to W.X.-Q. (20110071120093). The authors have no conflicts of interest to declare.
Databáze: OpenAIRE
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