C-Class CpG ODN: sequence requirements and characterization of immunostimulatory activities on mRNA level

Jurk, Marion; Schulte, Bettina; Kritzler, Andrea; Noll, Bernhard; Uhlmann, Eugen; Wader, Tanja; Schetter, Christian; Krieg, Arthur Μ.; Vollmer, Jörg

doi:10.1016/j.imbio.2004.02.006

Cited by 69 publications

(41 citation statements)

References 39 publications

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“…Our previous systematic analysis of the minimal sequence requirements for ODNs that activate hTLR9 (minH) confirmed that the 59TCG motif is essential for TLR9 activation but not sufficient (28), as indicated in the design of commonly used ODN2006 PTO (10,18,27,35,36). The ODNs with a 59TCG motif activate hTLR9 more effectively than mTLR9 as observed for the m109 and m127.…”

Section: Discussionmentioning

confidence: 60%

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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Section: Discussionmentioning

confidence: 60%

Species-Specific Minimal Sequence Motif for Oligodeoxyribonucleotides Activating Mouse TLR9

Pohar

Lainšček

Fukui

et al. 2015

The Journal of Immunology

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“…The 59-CG seems to be most important, but not sufficient, for TLR9 activation, as previously determined in the design of the generally used ODN2006 PTO and ODN2395 PTO (12,23,(29)(30)(31). The backbone modification to 29-O-methyl ribose within the 59 CpG motif, but not within the 39-CG-rich sequence, of the C-class ODN led to the complete loss of IFN-a secretion (30), confirming the importance of 59-CG and deoxyribose in the activation of TLR9.…”

Section: Discussionmentioning

confidence: 73%

“…The backbone modification to 29-O-methyl ribose within the 59 CpG motif, but not within the 39-CG-rich sequence, of the C-class ODN led to the complete loss of IFN-a secretion (30), confirming the importance of 59-CG and deoxyribose in the activation of TLR9. Moreover, CpG ODNs containing a 39 hairpin structure, but not the 59, are immunostimulatory (20), which is consistent with the finding that the accessible 59 CpG motif is necessary for activating TLR9.…”

Section: Discussionmentioning

confidence: 77%

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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“…Several well-studied ODN sequences were tested for their ability to induce Cox-2. It was previously shown that different sequences vary in their capacity to induce proliferation and cytokine production in human B cells [1,16,17]. Purified B cells were cultured separately with two different CpG sequences, ODN 2395 and ODN c274, or with a non-CpG sequence, GpC ODN 2137.…”

Section: Cpg Odn Induces Cox-2 Protein Expression In Human B Cellsmentioning

confidence: 99%

CpG oligodeoxynucleotides induce cyclooxygenase-2 in human B lymphocytes: Implications for adjuvant activity and antibody production

Bernard

Phipps

2007

Clinical Immunology

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Synthetic CpG oligodeoxynucleotides (ODN), similar to DNA sequences found in certain microorganisms, have shown promise as adjuvants for humans by enhancing immune responses. Since antibodies are often indicators of successful vaccination, it is important to understand how CpG ODN affects human B cells and influences antibody production. Treatment of human B cells with synthetic CpG ODN sequences increased both steady-state Cox-2 mRNA levels and protein expression. B cell receptor stimulation in concert with CpG ODN treatment induced Cox-2 expression and production of prostaglandin E 2 , well above that seen with CpG ODN alone. Importantly, CpG-induced human B cell IgM and IgG production was attenuated by dual Cox-1/ Cox-2 inhibitors and Cox-2 selective inhibitors. Our findings support a key role for CpG ODNinduced human B cell Cox-2 in the production of IgM and IgG antibodies, revealing that drugs that attenuate Cox-2 activity have the potential to reduce optimal antibody response to adjuvants/ vaccination.

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C-Class CpG ODN: sequence requirements and characterization of immunostimulatory activities on mRNA level

Cited by 69 publications

References 39 publications

Species-Specific Minimal Sequence Motif for Oligodeoxyribonucleotides Activating Mouse TLR9

Species-Specific Minimal Sequence Motif for Oligodeoxyribonucleotides Activating Mouse TLR9

Minimal Sequence Requirements for Oligodeoxyribonucleotides Activating Human TLR9

CpG oligodeoxynucleotides induce cyclooxygenase-2 in human B lymphocytes: Implications for adjuvant activity and antibody production

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