Research on molecular mechanisms of double stranded DNA-mediated innate immune activation

双链DNA介导先天免疫激活的分子机制研究

• 批准号: 18590432
• 负责人: TAKESHITA Fumihiko
• 金额: $ 2.44万
• 依托单位: Yokohama City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18590432/
• 关键词: double stranded DNA; Innate immunity; DNA virus infection; Histone; DNA vaccine; 感染免疫; シグナル伝達; 受容体

Research has been conducted 1)to identify molecules involved in innate immune activation mediated by double stranded DNA and 2)to analyze their roles in gene-mediated vaccines and gene therapies. By screening of a cDNA expression library, it was revealed that histone H2B, a component of the core histone complex, is involved in dsDNA (B-DNA)-mediated type I interferon (IFN) production pathway. We further examined about a role of histone H2B in innate immune responses by using RNA interference (siRNA) and found that nock-down of histone H2B led to suppression of type I IFN production induced by B-DNA. Moreover, knock-down of histone H2B resulted in elevation of replication efficiency of DNA viruses such as adenovirus, herpes simplex virus, and human papilloma virus, while it had little impact on replication of RNA viruses such as vesicular stomatitis virus and encephalomycarditis virus. These results suggested that histone H2B plays a significant role in recognition of viral DNA and initiation of innate immune responses to suppress viral replication. CIAO (C-terminal importin9-associated molecule organizing histone H2B and IPS-1) was identified after screening of histone H2B interacting molecules by using a yeast two hybrid system. It was shown that CIAO interacts with both histone H2B and IPS-1 and plays a critical role in IPS-1-mediated type I IFN production. Finally, it was demonstrated that the immunogenicity of DNA vaccine is critically controlled by an IRF kinase, TBK1, which is essential for histone H2B-mediated signaling by using TBK1 KO mice.

已进行的研究包括：1) 鉴定参与双链 DNA 介导的先天免疫激活的分子；2) 分析它们在基因介导的疫苗和基因治疗中的作用。通过筛选 cDNA 表达文库，发现核心组蛋白复合物的组成部分组蛋白 H2B 参与双链 DNA (B-DNA) 介导的 I 型干扰素 (IFN) 产生途径。我们通过使用 RNA 干扰 (siRNA) 进一步研究了组蛋白 H2B 在先天免疫反应中的作用，发现组蛋白 H2B 的沉默导致 B-DNA 诱导的 I 型 IFN 产生受到抑制。此外，敲低组蛋白H2B会导致腺病毒、单纯疱疹病毒、人乳头瘤病毒等DNA病毒的复制效率提高，而对水疱性口炎病毒、脑心肌炎病毒等RNA病毒的复制影响不大。这些结果表明组蛋白 H2B 在识别病毒 DNA 和启动先天免疫反应以抑制病毒复制方面发挥着重要作用。 CIAO（组织组蛋白 H2B 和 IPS-1 的 C 端导入相关分子）是在使用酵母二杂交系统筛选组蛋白 H2B 相互作用分子后鉴定的。结果表明，CIAO 与组蛋白 H2B 和 IPS-1 相互作用，并在 IPS-1 介导的 I 型 IFN 产生中发挥关键作用。最后，通过使用 TBK1 KO 小鼠，证明 DNA 疫苗的免疫原性受到 IRF 激酶 TBK1 的关键控制，TBK1 对于组蛋白 H2B 介导的信号传导至关重要。

项目成果

Histone H2B mediates intracellular recognition of naked DNA and production of type Iinterferons

组蛋白 H2B 介导细胞内裸露 DNA 的识别和 I 型干扰素的产生

• 发表时间: 2007
• 作者: Takeshita F;Kobiyama K;Suzuki K.
• 通讯作者: Suzuki K.

The Atg5-Atg12 conjugate associates with innate antiviral immune responses

• DOI: 10.1073/pnas.0704014104
• 发表时间: 2007-08-28
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Jounai, Nao;Takeshita, Fumihiko;Okuda, Kenji
• 通讯作者: Okuda, Kenji

Localization of C0R01A in the Macrophages Containing Mycobacterium leprae

C0R01A 在含有麻风分枝杆菌的巨噬细胞中的定位

• 发表时间: 2006
• 期刊: Acta. Histochem. Cytochem 39
• 作者: Suzuki;K.;Takeshita;F.;Nakata;N.;Ishii;N.;Makino;M
• 通讯作者: M

Blocking of the TLR5 activation domain hampers protective potential of DNA vaccine targeting flagellin

TLR5 激活域的阻断阻碍了针对鞭毛蛋白的 DNA 疫苗的保护潜力

• 发表时间: 2007
• 作者: Takeshita F;Saha S;Kobiyama K;Jounai N;Okuda K
• 通讯作者: Okuda K

Multivalent DNA vaccine protects mice against pulmonary infection caused by Pseudomonas aeruginosa

• DOI: 10.1016/j.vaccine.2006.05.077
• 发表时间: 2006-09-11
• 期刊: VACCINE
• 影响因子: 5.5
• 作者: Saha, Sukumar;Takeshita, Fumihiko;Okuda, Kenji
• 通讯作者: Okuda, Kenji