Toll-like receptor ligand activation of murine bone marrow-derived dendritic cells
Toll-like receptor ligand activation of murine bone marrow-derived dendritic cells
复制标题
DOI：
10.1111/j.1365-2567.2008.02922.x
复制
发表时间：
2009-04-01
期刊：
影响因子：
6.4
通讯作者：
Kimber, Ian
中科院分区：
文献类型：
--
作者：
Dearman, Rebecca J.;Cumberbatch, Marie;Kimber, Ian
树突状细胞（dc）是初级免疫反应启动所必需的。toll样受体（TLR）在这些细胞的不同亚群上的表达模式已被证明是不同的，这表明在影响免疫反应方面有不同的作用。我们在此研究了来自小鼠骨髓的未成熟dc （bmdc）对一系列TLR配体的反应。在粒细胞-巨噬细胞集落刺激因子存在下培养6天的bmdc，用TLR1-2 [Pam(3)Cys-Ser-(Lys)(4) (Pam)]、TLR2-6(巨噬细胞激活脂肽-2 (MALP-2)]配体刺激24小时；TLR3（多肌苷-多胞苷酸）、TLR4[脂多糖R515 (LPS)]、TLR5（鞭毛蛋白）、TLR7（多尿苷酸）和TLR9 [CpG ODN2395 (CpG)]。通过膜标记表达和细胞因子/趋化因子释放的培养上清分析来监测DC活化。TLR3和TLR7的配体无法激活BMDCs。其他TLR配体均引起膜标志物表达升高。PAM、MALP-2和LPS诱导促炎细胞因子和趋化因子的高水平表达。CpG治疗与1型细胞因子和趋化因子的偏好相关。Zymosan和PG具有促炎作用，但也偏向于2型细胞因子和趋化因子。相比之下，鞭毛蛋白没有引起BMDCs分泌显著的细胞因子或趋化因子。BMDCs的这些数据与报道的新分离小鼠朗格汉斯细胞的TLR库基本一致。此外，小鼠BMDCs对TLR配体的一般激活表现出选择性反应，分化的细胞因子模式提示可能引发不同的免疫反应。
Dendritic cells (DCs) are required for the initiation of primary immune responses. The pattern of Toll-like receptor (TLR) expression on various subsets of these cells has been shown to differ, suggestive of distinct roles in influencing immune responses. We have examined here the responses of immature DCs derived from murine bone marrow (BMDCs) to a range of TLR ligands. BMDCs cultured for 6 days in the presence of granulocyte-macrophage colony-stimulating factor were stimulated for 24 hr with ligands to TLR1-2 [Pam(3)Cys-Ser-(Lys)(4) (PAM)], TLR2-6 (macrophage-activating lipopeptide-2 (MALP-2); zymosan or peptidoglycan (PG)], TLR3 (polyinosinic-polycytidylic acid), TLR4 [lipopolysaccharide R515 (LPS)], TLR5 (flagellin), TLR7 (polyuridylic acid) and TLR9 [CpG ODN2395 (CpG)]. DC activation was monitored using membrane marker expression and analysis of culture supernatants for cytokine/chemokine release. Ligands to TLR3 and TLR7 failed to activate BMDCs. All other TLR ligands caused elevated expression of membrane markers. PAM, MALP-2 and LPS induced high-level expression of proinflammatory cytokines and chemokines. Treatment with CpG was associated with a preferential type 1 cytokine and chemokine profile. Zymosan and PG were proinflammatory but also skewed towards a type 2 pattern of cytokines and chemokines. In contrast, flagellin did not cause marked secretion by BMDCs of cytokines or chemokines. These data for BMDCs are largely consistent with the reported TLR repertoire of freshly isolated murine Langerhans cells. In addition, murine BMDCs show selective responses to TLR ligands with respect to general activation, with differentiated cytokine patterns suggestive of potential priming for divergent immune responses.