Cited by 55Immune Response and InflammationImmunotherapy and Immune ResponsesDNA and Nucleic Acid Chemistry

Minimal Sequence Requirements for Oligodeoxyribonucleotides Activating Human TLR9

Pohar, Jelka, Kužnik Krajnik, Alenka, Jerala, Roman, Benčina, Mojca·2015·The Journal of Immunology

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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Crossref · 10.4049/jimmunol.1402755

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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].”

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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
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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