Department of Biochemistry and Molecular Biology, Faculty of Medicine, Biomedical Center, University of Iceland, Iceland.
Faculty of Pharmaceutical Sciences, University of Iceland, Iceland.
Phytomedicine. 2015 Feb 15;22(2):277-82. doi: 10.1016/j.phymed.2014.12.007. Epub 2014 Dec 30.
Annotine is a lycopodane-type alkaloid isolated from the Icelandic club moss Lycopodium annotinum ssp. alpestre. Annotine does not inhibit acetylcholinesterase, as some other lycopodium alkaloids do, and other bioactivities have not been reported. The aim of this study was to determine the effects of annotine on maturation of dendritic cells (DCs) and their ability to activate allogeneic CD4(+) T cells. Human monocyte-derived DCs were matured in the absence or presence of annotine at a concentration of 1, 10 or 100 μg/ml. The effect of the annotine on maturation of the DCs was determined by measuring concentration of cytokines in culture supernatant by ELISA and expression of surface molecules by flow cytometry. DCs matured in the absence or presence of annotine at 100 µg/ml were also co-cultured with allogeneic CD4(+) T cells and concentration of cytokines in supernatants determined by ELISA and expression of surface molecules by flow cytometry. When cultured alone, DCs matured in the presence of annotine secreted less of the pro-inflammatory cytokines IL-6 and IL-23 and had a tendency toward less secretion of IL-12p40 than DCs matured in the absence of annotine. However, when DCs were matured in the presence of annotine and then co-cultured with allogeneic CD4(+) T cells they secreted more IL-12p40 and had a tendency toward secreting more IL-6 than DCs matured in the absence of annotine and then co-cultured with T cells. Allogeneic CD4(+) T cells co-cultured with DCs matured in the presence of annotine secreted more IL-13 than T cells co-cultured with DCs matured in the absence of annotine, but stimulating the DCs in the presence of annotine did not affect T cell secretion of IFN-γ and IL-17. There was also more IL-10 in co-cultures of T cells and DCs matured in the presence of annotine than in co-cultures of T cells and DCs matured in the absence of annotine. These results show that annotine increases the ability of DCs to direct the differentiation of allogeneic CD4(+) T cells toward a Th2/Treg phenotype, which may be of interest in the development of new treatments for Th1- and/or Th17-mediated inflammatory diseases.
annotine 是一种石松烷型生物碱，从冰岛石松 Lycopodium annotinum ssp. alpestre 中分离得到。与其他石松生物碱不同，annotine 不抑制乙酰胆碱酯酶，其他生物活性尚未报道。本研究旨在确定 annotine 对树突状细胞（DC）成熟及其激活同种异体 CD4(+)T 细胞能力的影响。用人单核细胞来源的 DC 在无 annotine 或浓度为 1、10 或 100μg/ml 的 annotine 存在下成熟。通过 ELISA 测定培养上清液中细胞因子浓度和流式细胞术测定表面分子表达来确定 annotine 对 DC 成熟的影响。无 annotine 或 100µg/ml 浓度 annotine 存在下成熟的 DC 也与同种异体 CD4(+)T 细胞共培养，通过 ELISA 测定上清液中细胞因子浓度和流式细胞术测定表面分子表达。单独培养时，与无 annotine 存在下成熟的 DC 相比，在 annotine 存在下成熟的 DC 分泌的促炎细胞因子 IL-6 和 IL-23 较少，IL-12p40 的分泌也有减少的趋势。然而，当 DC 在 annotine 存在下成熟并与同种异体 CD4(+)T 细胞共培养时，它们分泌的 IL-12p40 较多，与无 annotine 存在下成熟并与 T 细胞共培养的 DC 相比，它们分泌 IL-6 的趋势也较大。与在 annotine 存在下成熟的 DC 共培养的同种异体 CD4(+)T 细胞分泌的 IL-13 多于与无 annotine 存在下成熟的 DC 共培养的 T 细胞，而刺激 DC 在 annotine 存在下不会影响 T 细胞分泌 IFN-γ 和 IL-17。在 annotine 存在下成熟的 DC 与 T 细胞共培养的共培养物中也有更多的 IL-10，而在无 annotine 存在下成熟的 DC 与 T 细胞共培养的共培养物中则较少。这些结果表明，annotine 增加了 DC 引导同种异体 CD4(+)T 细胞向 Th2/Treg 表型分化的能力，这可能对开发针对 Th1 和/或 Th17 介导的炎症性疾病的新治疗方法具有重要意义。
