Funiculosin variants and phosphorylated derivatives promote innate immune responses via the Toll-like receptor 4/myeloid differentiation factor-2 complex-猫眼课题宝

DOI：

10.1074/jbc.m117.791780

发表时间：

2017-07

期刊：

The Journal of Biological Chemistry

影响因子：

通讯作者：

中科院分区：

其他

文献类型：

作者：

Naoki Okamoto;Keisuke Mizote;Hiroe Honda;A. Saeki;Yasuharu Watanabe;Tomomi Yamaguchi-Miyamoto;Ryutaro Fukui;N. Tanimura;Y. Motoi;Sachiko Akashi-Takamura;Tatsuhisa Kato;S. Fujishita;Takahito Kimura;U. Ohto;Toshiyuki Shimizu;T. Hirokawa;K. Miyake;K. Fukase;Y. Fujimoto;Y. Nagai;K. Takatsu

Toll样受体4（TLR 4）/髓样分化因子-2（MD-2）复合物对于LPS识别是必需的，并且诱导针对革兰氏阴性菌的先天性免疫应答。由于TLR 4/MD-2的活化对于诱导适应性免疫应答也是关键的，因此TLR 4/MD-2激动剂已被开发为疫苗佐剂，但其功效尚未确定。在这里，我们证明了一个funiculosin（FNC）的变体，FNC-RED，FNC-RED和FNC衍生物的激动剂为鼠和人TLR 4/MD-2。FNC-RED通过小鼠TLR 4/MD-2诱导核因子-κB（NF-κB）活化，而FNC没有TLR 4/MD-2刺激活性。Biacore分析显示，FNC-RED与鼠TLR 4/MD-2结合，但不与鼠辐射防护105（RP 105）/骨髓分化因子-1（MD-1）（另一种LPS传感器）结合。FNC-RED诱导小鼠巨噬细胞和树突状细胞中促炎细胞因子和共刺激分子的CD 14非依赖性表达。相反，FNC-RED刺激在CD 14依赖性LPS应答中减少，包括TLR 4/MD-2和IFN-β表达的二聚化和内化。FNC-RED诱导的小鼠树突状细胞IL-12 p40的产生依赖于NF-κB而非MAPK通路。此外，胎牛血清增强脂质A诱导的NF-κB活化，但阻断FNC-RED介导的反应。与FNC-RED不同，两种合成的含磷酸基团的FNC-RED和FNC衍生物FNC-RED-P01和FNC-P01分别激活人TLR 4/MD-2。最后，计算分析显示，FNC-RED和FNC-RED-P01的这种物种特异性激活是由鼠和人TLR 4/MD-2之间的静电表面电位差异引起的。我们得出结论，FNC-RED及其合成衍生物代表了一类新的小鼠和人类TLR 4/MD-2激动剂。

The Toll-like receptor 4 (TLR4)/myeloid differentiation factor-2 (MD-2) complex is essential for LPS recognition and induces innate immune responses against Gram-negative bacteria. As activation of TLR4/MD-2 is also critical for the induction of adaptive immune responses, TLR4/MD-2 agonists have been developed as vaccine adjuvants, but their efficacy has not yet been ascertained. Here, we demonstrate that a funiculosin (FNC) variant, FNC-RED, and FNC-RED and FNC derivatives are agonists for both murine and human TLR4/MD-2. FNC-RED induced nuclear factor-κB (NF-κB) activation via murine TLR4/MD-2, whereas FNC had no TLR4/MD-2 stimulatory activity. Biacore analysis revealed that FNC-RED binds to murine TLR4/MD-2 but not murine radioprotective 105 (RP105)/myeloid differentiation factor-1 (MD-1), another LPS sensor. FNC-RED induced CD14-independent expressions of pro-inflammatory cytokines and co-stimulatory molecules in murine macrophages and dendritic cells. In contrast, FNC-RED stimulation was reduced in CD14-dependent LPS responses, including dimerization and internalization of TLR4/MD-2 and IFN-β expression. FNC-RED-induced IL-12p40 production from murine dendritic cells was dependent on NF-κB but not MAPK pathway. In addition, fetal bovine serum augmented lipid A-induced NF-κB activation but blocked FNC-RED-mediated responses. Two synthetic phosphate group-containing FNC-RED and FNC derivatives, FNC-RED-P01 and FNC-P01, respectively, activated human TLR4/MD-2, unlike FNC-RED. Finally, computational analysis revealed that this species-specific activation by FNC-RED and FNC-RED-P01 resulted from differences in electrostatic surface potentials between murine and human TLR4/MD-2. We conclude that FNC-RED and its synthetic derivative represent a novel category of murine and human TLR4/MD-2 agonist.

Funiculosin variants and phosphorylated derivatives promote innate immune responses via the Toll-like receptor 4/myeloid differentiation factor-2 complex

文献摘要

相似文献