Minimal Sequence Requirements for Oligodeoxyribonucleotides Activating Human TLR9
Abstract
Synthetic oligodeoxyribonucleotides (ODNs) containing CpG (unmethylated deoxycytidylyl-deoxyguanosine dinucleotide) motifs activate endosomal TLR9. The nucleotide sequence, length, and dimerization properties of ODNs modulate their activation of TLR9. We performed a systematic investigation of the sequence motifs of B-class and C-class phosphodiester ODNs to identify the sequence properties that govern TLR9 activation. ODNs shorter than 21 nt and with the adenosine adjacent to the cytidine-guanosine (CG) dinucleotide motif led to a significant loss of the propensity to activate TLR9. The distance between the stimulatory CpG motifs within the ODN fine-tunes the activation of B cells. The minimal ODNs that activate human TLR9 comprise 2 CG dinucleotides separated by 6–10 nt, where the first CpG motif is preceded by the 5′-thymidine and the elongated poly-thymidine tail at the 3′ end of the ODN. The minimal sequence provides insight into the molecular mechanism of TLR9 ligand recognition. On the basis of sequence requirements, we conclude that two binding sites with different affinities for CG are formed in the human TLR9 dimer, with a very stringent binding site interacting with the 5′ CpG motif.
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Although GTCGTT has been proposed as an optimal human TLR9 activating sequence, other sequences of CpG ODN are also activating and questions remain regarding the potency of activation through sequence context, TLR secondary structure, and cell type.
“Although GTCGTT has been proposed as an optimal human TLR9 activating sequence, other sequences of CpG ODN are also activating and questions remain regarding the potency of activation through sequence context, TLR secondary structure, and cell type [96][97][98].”
From Risk mitigation of immunogenicity: a key to personalized retinal gene therapy (2021) · cites this paper as [97]
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Risk mitigation of immunogenicity: a key to personalized retinal gene therapy
Varin, J., Morival, C., Maillard, N., Adjali, O., & Cronin, T. (2021) · International Journal of Molecular Sciences · doi:10.3390/ijms222312818 · Cited by 19 · see citing works
- Authors
- Pohar, J., Kužnik Krajnik, A., Jerala, R., & Benčina, M.
- Affiliations
- National Institute of Chemistry (SI)
- Department of Biotechnology, National Institute of Chemistry , SI-1000 Ljubljana ,
- EN-FIST Centre of Excellence (Slovenia) (SI)
- Corresponding author
- Pohar, Jelka
- Jerala, Roman
- Venue
- The Journal of Immunology
- Publisher
- Oxford University Press (OUP)
- Published
- 2015-04
- Volume
- 194
- Issue
- 8
- Pages
- 3901-3908
- ISSN
- 0022-1767 (ISSN-L)
- 1550-6606
- OpenAlex
- W1742351369
- Cited by
- 55
- References
- 44
- Citations by year
- 2026: 3 · 2025: 5 · 2024: 3 · 2023: 2 · 2022: 6 · 2021: 5 · 2020: 3 · 2019: 3 · 2018: 10 · 2017: 8 · 2016: 5 · 2015: 2
- FWCI
- 4.10
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- The Journal of Immunology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › Immune Response and Inflammation
- Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA and Nucleic Acid Chemistry
- Keywords
- TLR9, CpG motifs, oligodeoxynucleotides, B cell activation
- MeSH terms
- B-Lymphocytes, Binding Sites, Humans, Oligodeoxyribonucleotides, Toll-Like Receptor 9, Protein Multimerization, HEK293 Cells
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