Recognition of CpG oligodeoxynucleotides by human Toll-like receptor 9 and subsequent cytokine induction

Suwarti, Suwarti; Yamazaki, Tomohiko; Svetlana, Chechetka; Hanagata, Nobutaka

doi:10.1016/j.bbrc.2012.12.068

Cited by 14 publications

(11 citation statements)

References 36 publications

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“…Introduction of the nucleaseresistant PTO backbone improved the ODN's uptake and hindered degradation by nucleases. A disadvantage of PTO modifications is the induction of an immune response independent of the CpG motif (42)(43)(44), which obscures the motif of the physiologically relevant agonists. The PTO-based minH ODN chimera comprising PD-based 59-TCGT maintained the ODN's species-specific specificity while retaining protection of the ODN against nucleases.…”

Section: Discussionmentioning

confidence: 99%

Minimal Sequence Requirements for Oligodeoxyribonucleotides Activating Human TLR9

Pohar

Krajnik

Jerala

et al. 2015

The Journal of Immunology

100

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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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Section: Discussionmentioning

confidence: 99%

Minimal Sequence Requirements for Oligodeoxyribonucleotides Activating Human TLR9

Pohar

Krajnik

Jerala

et al. 2015

The Journal of Immunology

100

View full text Add to dashboard Cite

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“…Therefore, a nuclease resistant PTO backbone was introduced, which improved the ODNs' uptake and hindered the nucleases. PTO modifications lead to the increased unspecific binding to proteins (41,42), and an induction of the immune response independent of the CpG motif and TLR9 (43)(44)(45)(46), which obscures the motif of the physiologically relevant agonists. The minH (28) and minM ODN chimeras comprising a 39 PTO-linked poly-T tail to hinder the nucleases and PD-linked motifs (59TCGT and 59TCCT 3 CG for the minH and minM, respectively) to maintain the ODNs' species-specific specificity are as efficient stimulators of immune cells as ODNs with a PTO backbone.…”

Section: Discussionmentioning

confidence: 99%

Species-Specific Minimal Sequence Motif for Oligodeoxyribonucleotides Activating Mouse TLR9

Pohar

Lainšček

Fukui

et al. 2015

The Journal of Immunology

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Synthetic oligodeoxyribonucleotides (ODNs) containing unmethylated CpG recapitulate the activation of TLR9 by microbial DNA. ODNs are potent stimulators of the immune response in cells expressing TLR9. Despite extensive use of mice as experimental animals in basic and applied immunological research, the key sequence determinants that govern the activation of mouse TLR9 by ODNs have not been well defined. We performed a systematic investigation of the sequence motif of B class phosphodiester ODNs to identify the sequence properties that govern mouse TLR9 activation. In contrast to ODNs activating human TLR9, where the minimal sequence motif for the receptor activation comprises a pair of closely positioned CpGs we found that the mouse TLR9 requires a single CpG positioned 4–6 nt from the 5′-end. Activation is augmented by a 5′TCC sequence one to three nucleotides from the CG. The distance of the CG dinucleotide of four to six nucleotides from the 5′-end and the ODN’s length fine-tunes activation of mouse macrophages. Length of the ODN <23 and >29 nt decreases activation of dendritic cells. The ODNs with minimal sequence induce Th1-type cytokine synthesis in dendritic cells and confirm the expression of cell surface markers in B cells. Identification of the minimal sequence provides an insight into the sequence selectivity of mouse TLR9 and points to the differences in the receptor selectivity between species probably as a result of differences in the receptor binding sites.

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“…In our series, significant positive correlations were observed between TLR9 and ZEB2, TWIST1 and VIM. Ultimately, as an interconnection was recently identified between TLR9 and EGFR pathways, we also explored EGFR at mRNA level and identified a We further investigated whether TLR9 overexpression by cancer epithelial cells was associated with TLR signaling pathway overexpression/activation [30]. By using real time quantitative RT-PCR in the TNC subgroup of IBCs, we confirmed that TNCs overexpressing TLR9 were characterized by activation of numerous genes implicated in TLR9 downstream signal transduction pathways: adaptors (MYD88), intracellular multimolecular mediators (IKBKE, IRAK1, IRAK3) and transcription factors (IRF7, IRF3, NFkB complex).…”

Section: Discussionmentioning

confidence: 99%

Biopathological Significance of TLR9 Expression in Cancer Cells and Tumor Microenvironment Across Invasive Breast Carcinomas Subtypes

Meseure

Vacher

Alsibai

et al. 2016

Cancer Microenvironment

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Toll-like receptors (TLRs) are pattern recognition receptors mainly expressed by cells of the immune system but also by epithelial tumor cells. Little is known about expression patterns of TLR genes in breast tumors, and their clinical significance is unclear. The aim of our study was to investigate expression of TLRs pathway components in pre-invasive breast lesions and invasive breast carcinomas (IBCs). We used RT-PCR assays to quantify mRNA levels of the 10 TLR genes and genes involved in TLR pathways in 350 breast tumors from patients with known clinical/pathological status and long-term outcome. Sets of 158 breast samples were also analyzed by immunochemistry including; 40 early noninvasive breast lesions, 38 IBCs and 80 triple negative carcinomas subtype (TNCs). We identified TLR9 as the major TLR gene family member upregulated in breast tumors and more particularly in TNCs. Immunohistochemical studies demonstrated that TLR9 protein was expressed in tumor epithelial and stromal cells of the TLR9 mRNA-overexpressing tumors. TLR9 overexpression appears very early during breast carcinogenesis. High TLR9 levels were associated with favorable outcome in the TNC sub-group. TLR9 overexpression was associated with alterations of down-stream components of the TLR9 signaling pathway, epithelio-mesenchymal transition (EMT) induction and EGFR pathway deregulation. TNCs with TLR9 overexpression were significantly correlated with development of a fibrous and inflammatory microenvironment with variable status of nuclear phosphoSTAT3. Our results suggest that TLR9 could play a role in TNC carcinogenesis and could be useful as predictive biomarker and therapeutic target.Keywords Toll-like receptor 9 . Breast cancer subtypes .

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Recognition of CpG oligodeoxynucleotides by human Toll-like receptor 9 and subsequent cytokine induction

Cited by 14 publications

References 36 publications

Minimal Sequence Requirements for Oligodeoxyribonucleotides Activating Human TLR9

Minimal Sequence Requirements for Oligodeoxyribonucleotides Activating Human TLR9

Species-Specific Minimal Sequence Motif for Oligodeoxyribonucleotides Activating Mouse TLR9

Biopathological Significance of TLR9 Expression in Cancer Cells and Tumor Microenvironment Across Invasive Breast Carcinomas Subtypes

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