Analyzing the biological significance in intracellular localization of Toll Like Receptors

分析 Toll 样受体细胞内定位的生物学意义

• 批准号: 21890050
• 负责人: SHIBATA Takuma
• 金额: $ 1.7万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Research Activity Start-up
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-21890050/
• 关键词: TLR4; LPS; TLR5; Flagellin; PRAT4A; マクロファージ; 細胞内分布; 自然免逸; Toll Like Receptor; 炎症; フラジェリン

Toll-like receptor s(TLRs) sense a variety of microbial products. TLRs activate innate immune responses and prime adaptive immune responses. TLR4/MD-2, a sensor for LPS, delivers the MyD88-dependent signal from the cellsurface, then traffics to endolysosomes and delivers the TRIF/TICAM-1-dependent signal. Both signals are thought to be dependent on cell surface TLR4/MD-2. Although TLR4/MD-2 is located also in recycling endosomes, the Golgi apparatus or the endoplasmic reticulum, little is known abouta role for intracellular TLR4/MD-2 in LPS responses. We here studied intracellular LPS sensing in macrophages. PRAT4A (protein associated with TLR4 A) is a cochaperone for a general chaperone gp96 and required for cell surface expression of TLR4/MD-2. Cell surface TLR4/MD-2 was undetectable on PRAT4A deficient thioglycollate-elicited peritoneal macrophage cells (P-Macs) and bone marrow-derived macrophages (BM-Macs). LPS responses were all abolished in PRAT4A deficient P-Macs, whereas a part of LPS responses remained detectable in PRAT4A deficient BM-Macs. Of note, LPS responses in PRAT4A deficient BM-Macs were not necessarily dependent on TRIF/TICAM-1 signaling. PRAT4A deficient BM-Macs showed unimpaired production of both TRIF/TICAM-1-dependent chemokine RANTES (CCL5) and MyD88-dependent chemokine MCP-1 (CCL2). Moreover, up-regulation of co-stimulatory molecules, CD40 and CD86 was notaltered. In contrast, TRIF/TICAM-1-dependent production of type I IFN was profoundly impaired. These results demonstrate that intracellular TLR4/MD-2 is responsible for unique set of LPS responses.

Toll样受体（TLR）可检测多种微生物产物。TLR激活先天免疫应答并引发适应性免疫应答。TLR 4/MD-2是LPS的传感器，从细胞表面传递MyD 88依赖的信号，然后运输到内溶酶体并传递TRIF/TICAM-1依赖的信号。这两种信号被认为依赖于细胞表面TLR 4/MD-2。尽管TLR 4/MD-2也位于再循环内体、高尔基体或内质网中，但关于胞内TLR 4/MD-2在LPS应答中的作用知之甚少。我们在这里研究了巨噬细胞中的细胞内LPS传感。PRAT 4A（与TLR 4A相关的蛋白质）是一般伴侣蛋白gp 96的共伴侣蛋白，并且是TLR 4/MD-2的细胞表面表达所需的。在PRAT 4A缺陷型巯基乙酸盐诱导的腹膜巨噬细胞（P-Macs）和骨髓源性巨噬细胞（BM-Macs）上检测不到细胞表面TLR 4/MD-2。在PRAT 4A缺陷的P-Macs中，LPS反应全部消失，而在PRAT 4A缺陷的BM-Macs中，仍可检测到部分LPS反应。值得注意的是，PRAT 4A缺陷型BM-Mac中的LPS应答不一定依赖于TRIF/TICAM-1信号传导。PRAT 4A缺陷BM-Mac显示TRIF/TICAM-1依赖性趋化因子RANTES（CCL 5）和MyD 88依赖性趋化因子MCP-1（CCL 2）的产生未受损。此外，共刺激分子CD 40和CD 86的上调也没有改变。相反，TRIF/TICAM-1依赖的I型IFN的产生受到严重损害。这些结果表明，细胞内TLR 4/MD-2负责独特的一组LPS应答。

项目成果

Unc93B1 restricts systemic lethal inflammation by orchestrating TLR7- and TLR9-trafficking

Unc93B1 通过协调 TLR7 和 TLR9 运输来限制全身性致命炎症

• 发表时间: 2011
• 期刊: Immunity誌 (in press)
• 作者: Ryutaro Fukui;Shin-ichiro Saitoh;Atsuo Kanno;Masahiro Onji;Takuma Shibata;Akihiko Ito;Morikazu Onji;Mitsuru Matsumoto;Shizuo Akira;Nobuaki Yoshida;Kensuke Miyake
• 通讯作者: Kensuke Miyake

Intarcellular TLR4/MD-2 in macrophages senses Gram-negative bacteria and induces a unique set of LPS-dependent genes

巨噬细胞中的细胞内 TLR4/MD-2 感知革兰氏阴性细菌并诱导一组独特的 LPS 依赖性基因

• 发表时间: 2011
• 期刊: International Immunology誌 (in press)
• 作者: Takuma Shibata;Yuji Motoi;Natsuko Tanimura;Natsuko Yamakawa;Sachiko Akashi-Takamura;Kensuke Miyake
• 通讯作者: Kensuke Miyake

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Toll Like Receptor 4 (TLR4)における細胞内分布の意義に関する研究

Toll样受体4（TLR4）细胞内分布意义的研究

• 发表时间: 2011
• 作者: 柴田琢磨;赤司(高村)祥子;三宅健介
• 通讯作者: 三宅健介