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树突状细胞(dendritic cell,DC)表面所表达的腺苷受体A2B亚型(ADOR-A2B)可促进DC对辅助性T淋巴细胞(T helper cell,Th)的激活,导致自身免疫性疾病的发生或加重.本文旨在研究作为免疫反应的诱导分子Toll样受体(Toll-like receptors,TLRs)是否可调节ADOR-A2B在DC中的表达并籍此影响其功能.体外诱导小鼠骨髓细胞分化为树突状细胞(BM-DC),以多种TLRs的配体,Pam3csk4、polyIC、LPS及CpG进行干预.提取细胞总RNA,real-time PCR测定ador-a2a、ador-a2b的表达;放射性配体结合实验测定BM-DC对3H-腺苷结合能力的变化.以LPS及选择性ADOR-A2B激动剂BAY 60-6583协同干预BM-DC,ELISA测定培养基中IL-1、IL-6及IL-12的含量.以干预后的BM-DC刺激naive CD4细胞,ELISA测定培养基中IL-17A、IFNγ的含量,荧光抗体染色及流式细胞仪分析检测CD4细胞的分化.结果显示,TLR-4的配体LPS可显著提高BM-DC中ador-a2b的表达及对腺苷的结合能力.BAY 60-6583与LPS相协同可刺激BM-DC分泌多种致炎因子,并增加其诱导CD4细胞向Th1及Th17分化的能力.由此可见,Toll样受体可上调adora2b在DC中的表达,并可籍此增加DC分泌促炎因子的能力及对CD4细胞的刺激作用.
Adenosine receptor subtype A2B (ADOR-A2B) expressed on the surface of dendritic cells (DCs) can promote the activation of T helper cells (Th) by DCs, resulting in autoimmune diseases The purpose of this paper is to investigate whether Toll-like receptors (TLRs), an inducing molecule of immune response, can regulate the expression of ADOR-A2B in DC and thereby affect its function.Methods: The cells were differentiated into dendritic cells (BM-DCs), which were intervened by various TLRs ligands, Pam3csk4, polyIC, LPS and CpG.The total RNA was extracted and the expression of ador-a2a and ador-a2b The binding of BM-DC to 3H-adenosine was determined by radioligand binding assay. BM-DCs were treated with LPS and selective ADOR-A2B agonist BAY 60-6583 in combination with IL-1, IL IL-6 and IL-12 in CD4 + CD25 + T cells were detected by flow cytometry.The naive CD4 cells were stimulated with BM-DC after intervention, the levels of IL-17A and IFNγ were detected by ELISA, the differentiation of CD4 cells was detected by fluorescent antibody and flow cytometry Showed that LPS, a ligand of TLR-4, significantly increased the expression of ador-a2b and the binding ability to adenosine in BM-DC.BAY 60-6583 and LP S can stimulate BM-DC to secrete a variety of inflammatory cytokines and increase the ability of CD4 cells to differentiate into Th1 and Th17 cells.Therefore, Toll-like receptors can upregulate the expression of adora2b in DC, Increase DC secretion of proinflammatory cytokines and stimulation of CD4 cells.