Cleavage of Toll-Like Receptor 9 Ectodomain Is Required for In Vivo Responses to Single Strand DNA
Mouse toll-like receptor 9 (TLR9) is an endosomal sensor for single-stranded DNA. TLR9 is transported from the endoplasmic reticulum to endolysosomes by a multiple transmembrane protein Unc93 homolog B1, and proteolytically cleaved at its ectodomain. The structure of TLR9 and its biochemical analyse...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-06-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/article/10.3389/fimmu.2018.01491/full |
| _version_ | 1818572220754034688 |
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| author | Ryutaro Fukui Chikako Yamamoto Fumi Matsumoto Masahiro Onji Masahiro Onji Takuma Shibata Yusuke Murakami Yusuke Murakami Atsuo Kanno Takuto Hayashi Natsuko Tanimura Nobuaki Yoshida Kensuke Miyake Kensuke Miyake |
| author_facet | Ryutaro Fukui Chikako Yamamoto Fumi Matsumoto Masahiro Onji Masahiro Onji Takuma Shibata Yusuke Murakami Yusuke Murakami Atsuo Kanno Takuto Hayashi Natsuko Tanimura Nobuaki Yoshida Kensuke Miyake Kensuke Miyake |
| author_sort | Ryutaro Fukui |
| collection | DOAJ |
| description | Mouse toll-like receptor 9 (TLR9) is an endosomal sensor for single-stranded DNA. TLR9 is transported from the endoplasmic reticulum to endolysosomes by a multiple transmembrane protein Unc93 homolog B1, and proteolytically cleaved at its ectodomain. The structure of TLR9 and its biochemical analyses have shown that the proteolytic cleavage of TLR9 ectodomain enables TLR9-dimerization and TLR9 activation. However, the requirement of TLR9 cleavage in vivo has not been studied. We here show that the 13 amino acids deletion at the cleavage site made TLR9 resistant to proteolytic cleavage. The deletion mutation in the Tlr9 gene impaired TLR9-dependent cytokine production in conventional dendritic cells from the mutant mice. Not only in vitro, in vivo production of inflammatory cytokines (TNF-α and IL-12p40), chemokine (CCR5/RANTES), and type I interferon (IFN-α) induced by administration of TLR9 ligand was also impaired. These results demonstrate that the TLR9 cleavage is required for TLR9 responses in vivo. |
| first_indexed | 2024-12-14T18:54:13Z |
| format | Article |
| id | doaj.art-098608ff306f4bcd8e80e8f7fd73df85 |
| institution | Directory Open Access Journal |
| issn | 1664-3224 |
| language | English |
| last_indexed | 2024-12-14T18:54:13Z |
| publishDate | 2018-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj.art-098608ff306f4bcd8e80e8f7fd73df852022-12-21T22:51:09ZengFrontiers Media S.A.Frontiers in Immunology1664-32242018-06-01910.3389/fimmu.2018.01491385746Cleavage of Toll-Like Receptor 9 Ectodomain Is Required for In Vivo Responses to Single Strand DNARyutaro Fukui0Chikako Yamamoto1Fumi Matsumoto2Masahiro Onji3Masahiro Onji4Takuma Shibata5Yusuke Murakami6Yusuke Murakami7Atsuo Kanno8Takuto Hayashi9Natsuko Tanimura10Nobuaki Yoshida11Kensuke Miyake12Kensuke Miyake13Division of Innate Immunity, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, JapanDivision of Innate Immunity, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, JapanDivision of Innate Immunity, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, JapanDivision of Innate Immunity, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, JapanInstitute of Molecular Biotechnology of the Austrian Academy of Sciences, Vienna, AustriaDivision of Innate Immunity, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, JapanDivision of Innate Immunity, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, JapanDepartment of Pharmacotherapy, Research Institute of Pharmaceutical Sciences, Musashino University, Tokyo, JapanDivision of Innate Immunity, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, JapanDivision of Innate Immunity, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, JapanDivision of Innate Immunity, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, JapanLaboratory of Developmental Genetics, The Institute of Medical Science, The University of Tokyo, Tokyo, JapanDivision of Innate Immunity, Department of Microbiology and Immunology, The Institute of Medical Science, The University of Tokyo, Tokyo, JapanLaboratory of Innate Immunity, Center for Experimental Medicine and Systems Biology, The Institute of Medical Science, The University of Tokyo, Tokyo, JapanMouse toll-like receptor 9 (TLR9) is an endosomal sensor for single-stranded DNA. TLR9 is transported from the endoplasmic reticulum to endolysosomes by a multiple transmembrane protein Unc93 homolog B1, and proteolytically cleaved at its ectodomain. The structure of TLR9 and its biochemical analyses have shown that the proteolytic cleavage of TLR9 ectodomain enables TLR9-dimerization and TLR9 activation. However, the requirement of TLR9 cleavage in vivo has not been studied. We here show that the 13 amino acids deletion at the cleavage site made TLR9 resistant to proteolytic cleavage. The deletion mutation in the Tlr9 gene impaired TLR9-dependent cytokine production in conventional dendritic cells from the mutant mice. Not only in vitro, in vivo production of inflammatory cytokines (TNF-α and IL-12p40), chemokine (CCR5/RANTES), and type I interferon (IFN-α) induced by administration of TLR9 ligand was also impaired. These results demonstrate that the TLR9 cleavage is required for TLR9 responses in vivo.https://www.frontiersin.org/article/10.3389/fimmu.2018.01491/fulltoll-like receptor 9proteolytic cleavageUnc93 homolog B1CpG-ODNprimary immune cellsin vivo response |
| spellingShingle | Ryutaro Fukui Chikako Yamamoto Fumi Matsumoto Masahiro Onji Masahiro Onji Takuma Shibata Yusuke Murakami Yusuke Murakami Atsuo Kanno Takuto Hayashi Natsuko Tanimura Nobuaki Yoshida Kensuke Miyake Kensuke Miyake Cleavage of Toll-Like Receptor 9 Ectodomain Is Required for In Vivo Responses to Single Strand DNA Frontiers in Immunology toll-like receptor 9 proteolytic cleavage Unc93 homolog B1 CpG-ODN primary immune cells in vivo response |
| title | Cleavage of Toll-Like Receptor 9 Ectodomain Is Required for In Vivo Responses to Single Strand DNA |
| title_full | Cleavage of Toll-Like Receptor 9 Ectodomain Is Required for In Vivo Responses to Single Strand DNA |
| title_fullStr | Cleavage of Toll-Like Receptor 9 Ectodomain Is Required for In Vivo Responses to Single Strand DNA |
| title_full_unstemmed | Cleavage of Toll-Like Receptor 9 Ectodomain Is Required for In Vivo Responses to Single Strand DNA |
| title_short | Cleavage of Toll-Like Receptor 9 Ectodomain Is Required for In Vivo Responses to Single Strand DNA |
| title_sort | cleavage of toll like receptor 9 ectodomain is required for in vivo responses to single strand dna |
| topic | toll-like receptor 9 proteolytic cleavage Unc93 homolog B1 CpG-ODN primary immune cells in vivo response |
| url | https://www.frontiersin.org/article/10.3389/fimmu.2018.01491/full |
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