Ding Yingjun, Cheng Xiang, Tang Tingting, Yao Rui, Chen Yong, Xie Jiangjiao, Yu Xian, Liao Yuhua
Institute of Cardiology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.
J Huazhong Univ Sci Technolog Med Sci. 2008 Aug;28(4):391-5. doi: 10.1007/s11596-008-0405-1. Epub 2008 Aug 15.
The purpose of the study was to observe the effect of rapamycin (RAPA) on the differentiation and maturation of rat bone marrow-derived dendritic cells (BMDCs) in vitro. BMDCs from Wistar rats were cultured with granulocyte-macrophage colony-stimulating factor plus interleukin-4 in the presence or absence of RAPA (20 ng/mL), and stimulated with lipopolysaccharide (LPS) for 24 h before cells and supernatants were collected. Surface phenotype of BMDCs was flow-cytometrically detected to determine the expression of maturation markers, MHC class II and CD86. Supernatants were analyzed for the production of IL-12 and IFN-gamma cytokines by using ELISA. BMDCs were co-cultured with T cells from Lewis rats and mixed lymphocyte reaction was assessed by MTT method. The morphology of BMDCs stimulated with LPS remained immature after RAPA pretreatment. RAPA significantly decreased the CD86 expression, impaired the IL-12 and IFN-gamma production of BMDCs stimulated with LPS, and inhibited the proliferation of allogeneic T cells. In conclusion, RAPA can inhibit the maturation of BMDCs stimulated with LPS in terms of the morphology, surface phenotype, cytokine production, and ability of BMDCs to stimulate the proliferation of allogeneic T cells in vitro.
本研究的目的是观察雷帕霉素(RAPA)对大鼠骨髓来源树突状细胞(BMDCs)体外分化和成熟的影响。将来自Wistar大鼠的BMDCs在存在或不存在RAPA(20 ng/mL)的情况下,用粒细胞-巨噬细胞集落刺激因子加白细胞介素-4进行培养,并在用脂多糖(LPS)刺激24小时后收集细胞和上清液。通过流式细胞术检测BMDCs的表面表型,以确定成熟标志物MHC II类和CD86的表达。使用ELISA分析上清液中IL-12和IFN-γ细胞因子的产生。将BMDCs与来自Lewis大鼠的T细胞共培养,并通过MTT法评估混合淋巴细胞反应。RAPA预处理后,用LPS刺激的BMDCs形态仍不成熟。RAPA显著降低CD86表达,损害用LPS刺激的BMDCs的IL-12和IFN-γ产生,并抑制同种异体T细胞的增殖。总之,就形态、表面表型、细胞因子产生以及BMDCs在体外刺激同种异体T细胞增殖的能力而言,RAPA可抑制用LPS刺激的BMDCs的成熟。