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Characterization of cooperative PS-oligo activation of human TLR9
Single-stranded phosphorothioate oligonucleotides (PS-oligos) can activate TLR9, leading to an innate immune response. This can occur with PS-oligos containing unmethylated CpG sites, the canonical motif, or PS-oligos that do not contain those motifs (non-CpG). Structural evidence shows that TLR9 co...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society of Gene & Cell Therapy
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10477407/ https://www.ncbi.nlm.nih.gov/pubmed/37675184 http://dx.doi.org/10.1016/j.omtn.2023.08.011 |
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author | Pollak, Adam J. Zhao, Luyi Crooke, Stanley T. |
author_facet | Pollak, Adam J. Zhao, Luyi Crooke, Stanley T. |
author_sort | Pollak, Adam J. |
collection | PubMed |
description | Single-stranded phosphorothioate oligonucleotides (PS-oligos) can activate TLR9, leading to an innate immune response. This can occur with PS-oligos containing unmethylated CpG sites, the canonical motif, or PS-oligos that do not contain those motifs (non-CpG). Structural evidence shows that TLR9 contains two PS-oligo binding sites, and recent data suggest that synergistic cooperative activation of TLR9 can be achieved by adding two separate PS-oligos to cells, each engaging with a separate site on TLR9 to enhance TLR9 activation as a pair. Here, we demonstrate and characterize this cooperativity phenomenon using PS-oligos in human cell lines, and we introduce several novel PS-oligo pairs (CpG and non-CpG pairs) that show cooperative activation. Indeed, we find that cooperative PS-oligos likely bind at different sites on TLR9. Interestingly, we find that PS-oligos that generate little TLR9 activation on their own can prime TLR9 to be activated by other PS-oligos. Finally, we determine that previous models of TLR9 activation cannot be used to fully explain data from systems using human TLR9 and PS-oligos. Overall, we reveal new details of TLR9 activation, but we also find that more work needs to be done to determine where certain PS-oligos are binding to TLR9. |
format | Online Article Text |
id | pubmed-10477407 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | American Society of Gene & Cell Therapy |
record_format | MEDLINE/PubMed |
spelling | pubmed-104774072023-09-06 Characterization of cooperative PS-oligo activation of human TLR9 Pollak, Adam J. Zhao, Luyi Crooke, Stanley T. Mol Ther Nucleic Acids Original Article Single-stranded phosphorothioate oligonucleotides (PS-oligos) can activate TLR9, leading to an innate immune response. This can occur with PS-oligos containing unmethylated CpG sites, the canonical motif, or PS-oligos that do not contain those motifs (non-CpG). Structural evidence shows that TLR9 contains two PS-oligo binding sites, and recent data suggest that synergistic cooperative activation of TLR9 can be achieved by adding two separate PS-oligos to cells, each engaging with a separate site on TLR9 to enhance TLR9 activation as a pair. Here, we demonstrate and characterize this cooperativity phenomenon using PS-oligos in human cell lines, and we introduce several novel PS-oligo pairs (CpG and non-CpG pairs) that show cooperative activation. Indeed, we find that cooperative PS-oligos likely bind at different sites on TLR9. Interestingly, we find that PS-oligos that generate little TLR9 activation on their own can prime TLR9 to be activated by other PS-oligos. Finally, we determine that previous models of TLR9 activation cannot be used to fully explain data from systems using human TLR9 and PS-oligos. Overall, we reveal new details of TLR9 activation, but we also find that more work needs to be done to determine where certain PS-oligos are binding to TLR9. American Society of Gene & Cell Therapy 2023-08-15 /pmc/articles/PMC10477407/ /pubmed/37675184 http://dx.doi.org/10.1016/j.omtn.2023.08.011 Text en © 2023 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Original Article Pollak, Adam J. Zhao, Luyi Crooke, Stanley T. Characterization of cooperative PS-oligo activation of human TLR9 |
title | Characterization of cooperative PS-oligo activation of human TLR9 |
title_full | Characterization of cooperative PS-oligo activation of human TLR9 |
title_fullStr | Characterization of cooperative PS-oligo activation of human TLR9 |
title_full_unstemmed | Characterization of cooperative PS-oligo activation of human TLR9 |
title_short | Characterization of cooperative PS-oligo activation of human TLR9 |
title_sort | characterization of cooperative ps-oligo activation of human tlr9 |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10477407/ https://www.ncbi.nlm.nih.gov/pubmed/37675184 http://dx.doi.org/10.1016/j.omtn.2023.08.011 |
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