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

• Project/Area Number: 18590432
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Bacteriology (including Mycology)
• Research Institution: Yokohama City University
• Principal Investigator: TAKESHITA Fumihiko Yokohama City University, Graduate School of Medicine, Associate Professor (60333572)
• Co-Investigator(Kenkyū-buntansha): SUZUKI Koichi National Institute of Infectious Diseases, Department of Bioregulation, Laboratory Chief (20206478)
• Project Period (FY): 2006 – 2007
• Project Status: Completed (Fiscal Year 2007)
• Budget Amount: ¥3,810,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥210,000); Fiscal Year 2007: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2006: ¥2,900,000 (Direct Cost: ¥2,900,000)
• Keywords: 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 Abstract

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.

Research Products

• [Journal Article] Tank-binding kinase-I delineates innate and adaptive immune responses raised by DNA vaccines (2008)
  • Author(s): Ishii KJ, Kawagoe T, Koyama S, Matsui K, Ktunar H, Kawai T, Uematsu S, Takeuchi O, Takeshita F, Coban C, and Akira S.
  • Journal Title: Nature 451 Pages: 725-728
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] The non-canonical role of Atg family members as suppressors of innate antiviral immune signaling (2008)
  • Author(s): Takeshita F, Kobiyama K, Miyawaki A, Jounai N, and Okuda K.
  • Journal Title: Autophagy 4 Pages: 1-4
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Tank-binding kinase-1 delineates innate and adaptive immune responses raised by DNA vaccines (2008)
  • Author(s): Ishii, KJ., Kawagoe, T., Koyama, S., Matsui, K., Kumar, H., Kawai, T., Uematsu, S., Takeuchi, O., Takeshita, F., Coban, C., Akira, S
  • Journal Title: Nature 451 Pages: 725-725
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The non-canonical role of Atg family members as suppressors of innate antiviral immune signaling (2008)
  • Author(s): Takeshita, F., Kobiyama, K., Miyawaki ,A., Jounai, N., Okuda, K
  • Journal Title: Autophagy 4 Pages: 1-4
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Tank-binding kinase-1 delineates innate and adaptive immune responses raised by DNA vaccines. (2008)
  • Author(s): Ishii KJ, Kawagoe T, Koyama S, Matsui K, Kumar H, Kawai T, Uematsu S, Takeuchi O, Takeshita F, Coban C, and Akira S.
  • Journal Title: Nature 451 Pages: 725-728
  • Peer Reviewed
• [Journal Article] The non-canonical role of Atg family members as suppressors of innate antiviral immune signaling. (2008)
  • Author(s): Takeshita F, Kobiyama K, Miyawaki A, Jounai N, and Okuda K.
  • Journal Title: Autophagy 4 Pages: 1-4
  • Peer Reviewed
• [Journal Article] The Atg5-Atg12 conjugate associates with innate antiviral immune responses (2007)
  • Author(s): Jounai N, Takeshita F, Kobiyama K, Sawano A, Miyawaki A, Xin KQ, Ishii KJ, Kawai T, Akira S, Suzuki K, and Okuda K.
  • Journal Title: Proceedings National Academy of Science USA 104 Pages: 14050-14050
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Blocking of the TLR5 activation domain hampers protective potential of DNA vaccine targeting flagellin (2007)
  • Author(s): Saha S, Takeshita F, Matsuda T, Jounai N, Kobiyama K, Matsumoto T, Sasaki S, Yoshida A, Xin KQ, Klinman DM, Uematsu S, Ishii KJ, Akira S, and Okuda K.
  • Journal Title: The Journal of Immunology 179 Pages: 1147-1154
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] TLR9 gexpression and functionis abolished by the cervical cancer-assocfated human papillomavirus type 16 (2007)
  • Author(s): Hasan UA, Bates E, Takeshita F, Biliato A, Accardi R, Bouvard V, Mansour M, Vincent I, Gissmann L, Iftner T, Sideri M, Stubenrauch F, and Tommasino M.
  • Journal Title: The Journal of Immunology 178 Pages: 3186-3197
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Mutant influenza A virus nucleoprotein is preferentially localized in the cytoplasm and its immunization in mice shows higher immunogenicity and cross-reactivity (2007)
  • Author(s): Ohba K, Yoshida S, Zahidunnabi Dewan M, Shimura H, Sakamaki N, Takeshita F, Yamamoto N, and Okuda K.
  • Journal Title: Vaccine 25 Pages: 4291-4300
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Suppression of iodide uptake and thyroid hormone synthesis with stimulation of the type I interferon system by double-stranded RNA(dsRNA) in cultured human thyroid follicles (2007)
  • Author(s): Yamazaki K, Suzuki K, Yamada E, Yamada T, Takeshita F, Matsumoto M, Mitsuhashi T, Obara T, Takano K, and Sato K.
  • Journal Title: Endocrinology 148 Pages: 13226-3235
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] The degree of apoptosis as an immunostimulant for a DNA vaccine against HIV-1 infection (2007)
  • Author(s): Kojima Y, Jounai N, Takeshita F, Nakazawa M, Okuda K, Watabe S, Xin KQ, Okuda K.
  • Journal Title: Vaccine 25 Pages: 438-445
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Muscle creatine kinase/SV 40 hybrid promoter for muscle-targeted long-term transgene expression (2007)
  • Author(s): Takeshita F, Takase K, Tozuka M, Saha S, Okuda K, Ishii N, and Sasaki S
  • Journal Title: International Journal of Molecular Medicine 19 Pages: 309-335
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] The Atg5-Atgl2 conjugate associates with innate antiviral immune responses (2007)
  • Author(s): Jounai, N., Takeshita, F., Kobiyama, K., Sawano, A., Miyawaki, A., Xin KQ., Ishii ,KJ., Kawai, T., Akira, S., Suzuki, K., Okuda, K
  • Journal Title: Proc. Natl, Acad. Sci. USA 104 Pages: 14050-14055
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Blocking of the TLR5 activation domain hampers protective potential of DNA vaccine targeting flagellin (2007)
  • Author(s): Saha, S., Takeshita, F., Matsuda, T., Jounai, N., Kobiyama, K., Matsumoto, T., Sasaki, S., Yoshida, A., Xin, KQ., Klinman, DM., Uematsu, S., Ishii, KJ., Akira, S., Okuda, K
  • Journal Title: J. Immunol 179 Pages: 1147-1154
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] TLR9 expression and function is abolished by the cervical cancer-associated human papillomavirus type 16 (2007)
  • Author(s): Hasan ,UA., Bates, E., Takeshita, F., Biliato, A., Accardi, R., Bouvard, V., Mansour, M., Vincent, I., Gissmann, L., Iftner, T., Sideri, M., Stubenrauch, F., Tommasino, M
  • Journal Title: J. Immunol 178 Pages: 3186-3197
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Mutant influenza A virus nucleoprotein is preferentially localized in the cytoplasm and its immunization in mice shows higher immunogenicity and cross-reactivity (2007)
  • Author(s): Ohba, K., Yoshida, S., Zahidunnabi, Dewan, M., Shimura, H., Sakamaki, N., Takeshita, F., Yamamoto, N., Okuda ,K
  • Journal Title: Vaccine 25 Pages: 4291-4300
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Suppression of iodide uptake and thyroid hormone synthesis with stimulation of the type I interferon system by double-stranded RNA(dsRNA)in cultured human thyroid follicles (2007)
  • Author(s): Yamazaki, K., Suzuki, K., Yamada, E., Yamada, T., Takeshita, F., Matsumoto, M., Mitsuhashi, T., Obara, T., Takano, K., Sato, K
  • Journal Title: Endocrinology 148 Pages: 3226-3235
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Muscle creatine kinase/SV 40 hybrid promoter for muscle-targeted long-term transgene expression (2007)
  • Author(s): Takeshita, F., Takase, K., Tozuka, M., Saha, S., Okuda, K., Ishii, N., Sasaki, S
  • Journal Title: Int. J. Mol. Med 19 Pages: 309-315
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The degree of apoptosis as an immunostimulant for a DNA vaccine against HIV-1 infection (2007)
  • Author(s): Kojima, Y., Jounai, N., Takeshita, F., Nakazawa, M., Okuda, K., Watabe, S., Xin, KQ., Okuda, K
  • Journal Title: Vaccine 25 Pages: 438-445
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The Atg5-Atg12 conjugate associates with innate antiviral immune responses. (2007)
  • Author(s): Jounai N, Takeshita F. Kobiyama K, Sawano A, Miyawaki A, Xin KQ, Ishii KJ, Kawai T, Akira S, Suzuki K, and Okuda K.
  • Journal Title: Proceedings National Academy of Science USA 104 Pages: 14050-14050
  • Peer Reviewed
• [Journal Article] Blocking of the TLR5 activation domain hampers protective potential of DNA vaccine targeting flagellin. (2007)
  • Author(s): Saha S, Takeshita F. Matsuda T, Jounai N, Kobiyama K, Matsumoto T, Sasaki S, Yoshida A, Xin KQ, Klinman DM, Uematsu S, Ishii KJ, Akira S, and Okuda K.
  • Journal Title: The Journal of Immunology 179 Pages: 1147-1154
  • Peer Reviewed
• [Journal Article] TLR9 expression and function is abolished by the cervical cancer-associated human papillomavirus type 16. (2007)
  • Author(s): Hasan UA, Bates E, Takeshita F, Biliato A, Accardi R, Bouvard V, Mansour M, Vincent I, Gissmann L, Iftner T, Sideri M, Stubenrauch F, and Tommasino M.
  • Journal Title: The Journal of Immunology 178 Pages: 3186-3197
  • Peer Reviewed
• [Journal Article] Mutant influenza A virus nucleoprotein is preferentially localized in the cytoplasm and its immunization in mice shows higher immunogenicity and cross-reactivity. (2007)
  • Author(s): Ohba K, Yoshida S, Zahidunnabi Dewan M, Shimura H, Sakamaki N, Takeshita F, Yamamoto N, and Okuda K.
  • Journal Title: Vaccine 25 Pages: 4291-4300
  • Peer Reviewed
• [Journal Article] Suppression of iodide uptake and thyroid hormone synthesis with stimulation of the type I interferon system by double-stranded RNA(dsRNA)in cultured human thyroid follicles. (2007)
  • Author(s): Yamazaki K, Suzuki K, Yamada E, Yamada T, Takeshita F, Matsumoto M, Mitsuhashi T, Obara T, Takano K and Sato K
  • Journal Title: Endocrinology 148 Pages: 13226-3235
  • Peer Reviewed
• [Journal Article] The degree of apoptosis as an immunostimulant for a DNA vaccine against HIV-1 infection. (2007)
  • Author(s): Kojima Y, Jounai N, Takeshita F, Nakazawa M, Okuda K, Watabe S, Xin KQ, Okuda K.
  • Journal Title: Vaccine 25 Pages: 438-445
  • Peer Reviewed
• [Journal Article] Muscle creatine kinase/SV 40 hybrid promoter for muscle-targeted long-term transgene expression. (2007)
  • Author(s): Takeshita F, Takase K, Tozuka M, Saha S, Okuda K, Ishii N, and Sasaki S
  • Journal Title: International Journal of Molecular Medicine 19 Pages: 309-315
  • Peer Reviewed
• [Journal Article] Suppression of iodite uptake and thyroid hormone synthesis with stimulation of the type I interferon system by double-stranded RNA (dsRNA) in cultured human thyroid follicles. (2007)
  • Author(s): Yamazaki K, Suzuki K, Yamada E, Yamada T, Takeshita F, Matsumoto M, Mitsuhashi T, Obara T, Takano K, Sato K
  • Journal Title: Endocrinology (in press)
• [Journal Article] Muscle creatine kinase/SV 40 hybrid promoter for muscle-targeted long-term transgene expression (2007)
  • Author(s): Takeshita F, Takase K, Tozuka M, Saha S, Okuda K, Ishii N, Sasaki S
  • Journal Title: Int. J. Mol. Med. 19 Pages: 309-315
• [Journal Article] A fused gene of nucleoprotein(NP) and herpes simplex virus genes(VP22) induces highly protective immunity against different subtypes of influenza virus (2006)
  • Author(s): Saha S, Yoshida S, Ohba K, Matsui K, Matsuda T, Takeshita F, Umeda K, Tamura Y, Okuda K, Klinman D, Xin KQ, and Okuda K
  • Journal Title: Virology 35 Pages: 48-57
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Multivalent DNA vaccine protects mice against pulmonary infection caused by Pseudomonas aeruginosa (2006)
  • Author(s): Saha S, Tkeyshims F, Sasald S, Matsuda T, Tanaka T, Tozuka M, Takase K, Matsumoto T, Okuda K, Ishii N, Yamaguchi K, Xin KQ, Klinman DM, and Okuda K
  • Journal Title: Vaccine 24 Pages: 6240-6249
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Activator protein 1-mediated transcriptional regulations strategy sustains long-term expression of a xenogeneic gene product invivo: An implication for gene therapy tareeting coneenital protein deficiencies (2006)
  • Author(s): Sasaki S, Smith JM, Takase K, Okuda K, Ishii N, and Takeshita F
  • Journal Title: International Journal of Molecular Medicine 18 Pages: 289-297
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] TLR-adaptor molecules enhance DNA-raised adaptive immune responses against flu and tumor challenge through the activation of innate immunity (2006)
  • Author(s): Takeshita F, Tanaka T, Matsuda T, Tozuka M, Kobiyama K, Saha S, Matsui K, Ishii KJ, Coban C, Akira S, Ishii N, Suzuki K, Klinman DM, Okuda K, and Sasai S
  • Journal Title: The Journal of Virology 80 Pages: 6218-6224
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Localization of C0R01A in the Macrophages Containing Mycobacterium leprae (2006)
  • Author(s): Suzuki, K., Takeshita, F., Nakata, N., Ishii, N., Makino, M
  • Journal Title: Acta. Histochem. Cytochem 39 Pages: 107-112
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A fused gene of nucleoprotein(NP)and herpes simplex virus genes(VP22)induces highly protective immunity against different subtypes of influenza virus (2006)
  • Author(s): Saha, S., Yoshida, S., Ohba, K., Matsui, K., Matsuda, T., Takeshita, F., Umeda, K., Tamura, Y., Okuda, K., Klinman D., Xin, KQ., Okuda, K
  • Journal Title: Virology 35 Pages: 48-57
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Multivalent DNA vaccine protects mice against pulmonary infection caused by Pseudomonas aeruginosa (2006)
  • Author(s): Saha, S., Takeshita, F., Sasaki, S., Matsuda, T., Tanaka, T., Tozuka, M., Takase, K., Matsumoto, T., Okuda, K., Ishii, N., Yamaguchi, K., Xin, KQ., Klinman, DM., Okuda, K
  • Journal Title: Vaccine 24 Pages: 6240-6249
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Activator protein 1-mediated transcriptional regulation strategy sustains long-term expression of a xenogeneic gene product in vivo: An implication for gene therapy targeting congenital protein deficiencies (2006)
  • Author(s): Sasaki, S., Smith, JM., Takase, K., Okuda, K., Ishii, N., Takeshita, F
  • Journal Title: Int. J. Mol. Med 18 Pages: 289-297
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Saha (2006)
  • Author(s): Takeshita, F., Tanaka, T., Matsuda, T., Tozuka, M., Kobiyama, K
  • Journal Title: J. Virol 80 Pages: 6218-6224
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] A fused gene of nucleoprotein (NP) and herpes simplex virus genes (VP22) induces highly protective immunity against different subtypes of influenza virus (2006)
  • Author(s): Saha S, Yoshida S, Ohba K, Matsui K, Matsuda T, Takeshita F, Umeda K, Tamura Y, Okuda K, Klinman D, Xin KQ, Okuda K
  • Journal Title: Virology 35 Pages: 48-57
• [Journal Article] Multivalent DNA vaccine protects mice against pulmonary infection caused by Pseudomonas aeruginosa (2006)
  • Author(s): Saha S, Takeshita F, Sasaki S, Matsuda T, Tanaka T, Tozuka M, Takase K, Matsumoto T, Okuda K, Ishii N, Yamaguchi K, Xin KQ, Klinman DM, Okuda K
  • Journal Title: Vaccine 24 Pages: 6240-6249
• [Journal Article] Activator protein 1-mediated transcriptional regulation strategy sustains long-term expression of a xenogeneic gene product in vivo : An implication for gene therapy targeting congenital protein deficiencies (2006)
  • Author(s): Sasaki S, Smith JM, Takase K, Okuda K, Ishii N, Takeshita F
  • Journal Title: Int. J. Mol. Med. 18 Pages: 289-297
• [Journal Article] TLR-adaptor molecules enhance DNA-raised adaptive immune responses against flu and tumor challenge through the activation of innate immunity (2006)
  • Author(s): Takeshita F, Tanaka T, Matsuda T, Tozuka M, Kobiyama K, Saha S, Matsui K, Ishii KJ, Coban C, Akira S, Ishii N, Suzuki K, Klinman DM, Okuda K, Sasai S
  • Journal Title: J. Virol 80 Pages: 6218-6224
• [Presentation] Histone H2B mediates intracellular recognition of naked DNA and production of type Iinterferons (2007)
  • Author(s): Takeshita F, Kobiyama K, Suzuki K.
  • Organizer: 第30回目本分子生物学会
  • Place of Presentation: 横浜
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Blocking of the TLR5 activation domain hampers protective potential of DNA vaccine targeting flagellin (2007)
  • Author(s): Takeshita F, Saha S, Kobiyama K, Jounai N, and Okuda K
  • Organizer: 第20回内藤カンファランス
  • Place of Presentation: 葉山
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Atg family members associate with theds RNA-mediated innate anti-virusimmune pathway (2007)
  • Author(s): 武王 文彦、城内 直、小檜山 康司、奥田 研爾
  • Organizer: 第80回日本細菌学会総会
  • Place of Presentation: 大阪
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Autophagy-related Atg family members associate with the dsRNA-mediated innate anti-virus immune pathway (2007)
  • Author(s): Kobiyama K, Jounai N, and Takeshita F
  • Organizer: Keystone Symposiaon Molecular and Cellular Biology
  • Place of Presentation: Keystone,Colorado
  • Description: 「研究成果報告書概要(和文)」より
• [Presentation] Histone H2B mediates intracellular recognition of naked DNA and production of tvoe I interferons (2007)
  • Author(s): Takeshita, F., Kobiyama, K., Suzuki, K
  • Organizer: The 30th Annual Meeting of the Molecular Biology Society of Japan
  • Place of Presentation: Yokohama
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Blocking of the TLR5 activation domain hampers protective potential of DNA vaccine targeting flagellin (2007)
  • Author(s): Takeshita, F., Saha, S., Kobiyama, K., Joubai, N., Okuda, K
  • Organizer: The 20th Naito Conference On Innate Immunity in Medicine and Biology III
  • Place of Presentation: Hayama
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Atg family members associate with the dsRNA-mediated innate antivirus immune pathway (2007)
  • Author(s): Takeshita, F., Jounai, N., Kobiyama, K., Okuda, K
  • Organizer: The 80th Annual Meeting of the Japanese Society for Bacteriology
  • Place of Presentation: Osaka
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Autophagy-related Atg family members associate with the dsRNA-mediated innate anti-virus immune pathway (2007)
  • Author(s): Kobiyama, K., Jounai, N., Takeshita F
  • Organizer: Keystone Symposia on Molecular and Cellular Biology
  • Place of Presentation: Keystone, CO, USA
  • Description: 「研究成果報告書概要(欧文)」より
• [Presentation] Histone H2B mediates intracellular recognition of naked DNA and production of type I interferons (2007)
  • Author(s): Takeshita F, Kobiyama K, Suzuki K.
  • Organizer: 第30回日本分子生物学会
  • Place of Presentation: 横浜
• [Presentation] Blocking of the TLR5 activation domain hampers protective potential of DNA vaccine targeting flagellin. (2007)
  • Author(s): Takeshita F, Saha S, Kobiyama K, Jounai N, and Okuda K
  • Organizer: 第20回内藤カンファランス
  • Place of Presentation: 葉山