Oligonucleotide Sequences Required for Natural Killer Cell Activation

Kuramoto, Etsuro; Yano, Okihito; Kimura, Yasuto; Baba, Makoto; Makino, T.; Yamamoto, Saburo; Yamamoto, Toshiko; Kataoka, Tateshi; Tokunaga, Tohru

doi:10.1111/j.1349-7006.1992.tb02734.x

Cited by 117 publications

(45 citation statements)

References 14 publications

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“…Such studies have included both in vitro assays and, to a lesser extent, in vivo animal models. Furthermore, it has been suggested that the antitumor activity exerted by some phosphorothioate ODNs may be due to either a non-sequence-related effect of ODNs on binding to specific growth factors or a sequencespecific effect of ODNs on stimulation of natural killer cell activity in host animals (16)(17)(18)(19)(20). These reports, as well as others (2,14,15), demonstrate the need to rigorously examine the mechanism of action of a particular ODN before concluding that the pharmacological effects exerted by that ODN are derived from an antisense mechanism of action.…”

Section: Discussionmentioning

confidence: 99%

Sequence-specific antitumor activity of a phosphorothioate oligodeoxyribonucleotide targeted to human C- raf kinase supports an antisense mechanism of action in vivo

Monia

Sasmor

Johnston

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

132

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To determine the mechanism of action responsible for the in vivo antitumor activity of a phosphorothioate antisense inhibitor targeted against human C-raf kinase (ISIS 5132, also known as CGP69846A), a series of mismatched phosphorothioate analogs of ISIS 5132 or CGP69846A were synthesized and characterized with respect to hybridization affinity, inhibitory effects on C-raf gene expression in vitro, and antitumor activity in vivo. Incorporation of a single mismatch into the sequence of ISIS 5132 or CGP69846A resulted in reduced hybridization affinity toward C-raf RNA sequences and reduced inhibitory activity against C-raf expression in vitro and tumor growth in vivo. Moreover, incorporation of additional mismatches resulted in further loss of in vitro and in vivo activity in a manner that correlated well with a hybridization-based (i.e., antisense) mechanism of action. These results provide important experimental evidence supporting an antisense mechanism of action underlying the in vivo antitumor activity displayed by ISIS 5132 or CGP69846A.Unequivocal proof of the in vivo mechanism of action for any drug has always been one of the greatest challenges facing the molecular pharmacologist. This challenge is mainly due to the difficulty in obtaining sound experimental evidence for drug mechanisms in complex animal models or in humans. Indeed, the mechanisms of action for many drugs that are used routinely in the clinic today with significant therapeutic benefit are poorly characterized. Unfortunately, without a clear understanding of a particular drug's mechanism in vivo, improvement in the drug's pharmacological properties and advances in the discovery of new drug classes based on novel mechanisms of action will be greatly impeded.Antisense oligonucleotides represent a new paradigm for drug discovery that holds great promise for the delivery of potent and specific drugs with fewer undesired side effects (reviewed in refs. 1 and 2). This paradigm offers the opportunity to rapidly identify lead compounds based upon knowledge of the biology of a disease process. With this knowledge, the practitioner of antisense drug discovery can rapidly design, synthesize, and test a series of compounds in animal models and, because of the rational manner by which these compounds are designed to act, can experimentally address their mechanism of action in vivo with a relatively high degree of confidence. The most compelling evidence to support antisense mechanisms in living systems are (i) specific reduction in target gene expression and (ii) gradual reduction in drug potency as a result of incorporating increasing numbers of nonhybridizing bases (mismatches) into the oligonucleotide (i.e., rank order potency based on hybridization affinity). However, despite the fact that oligonucleotides have been shown to exert potent pharmacological activity in a variety of animal models, rigorous analyses providing strong experimental evidence to support an antisense mechanism of action underlying in vivo pharmacological activity have...

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Section: Discussionmentioning

confidence: 99%

Sequence-specific antitumor activity of a phosphorothioate oligodeoxyribonucleotide targeted to human C- raf kinase supports an antisense mechanism of action in vivo

Monia

Sasmor

Johnston

et al. 1996

Proc. Natl. Acad. Sci. U.S.A.

132

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“…Because the GACGATCGTC sequence exists in the bacterial genomes, with various numbers of G on each side, we used the pal-GACGA1010 in unmodified phosphodiester form as a counterpart of bacterial DNA, which has previously been shown to have immunostimulatory activity in mice and humans (14,32).…”

Section: Cpg Dna Induces Human Pdcs To Express Cxcl10 and Ccl3 In Addmentioning

confidence: 99%

Collaborative Action of NF-κB and p38 MAPK Is Involved in CpG DNA-Induced IFN-α and Chemokine Production in Human Plasmacytoid Dendritic Cells

Osawa¹,

Iho

Takauji³

et al. 2006

The Journal of Immunology

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CpG DNA induces plasmacytoid dendritic cells (pDC) to produce type I IFN and chemokines. However, it has not been fully elucidated how the TLR9 signaling pathway is linked to these gene expressions. We examined the mechanisms involving the TLR9 and type I IFN signaling pathways, in relation to CpG DNA-induced IFN-α, IFN regulatory factor (IRF)-7, and chemokines CXCL10 and CCL3 in human pDC. In pDC, NF-κB subunits p65 and p50 were constitutively activated. pDC also constitutively expressed IRF-7 and CCL3, and the gene expressions seemed to be regulated by NF-κB. CpG DNA enhanced the NF-κB p65/p50 activity, which collaborated with p38 MAPK to up-regulate the expressions of IRF-7, CXCL10, and CCL3 in a manner independent of type I IFN signaling. We then examined the pathway through which IFN-α is expressed. Type I IFN induced the expression of IRF-7, but not of IFN-α, in a NF-κB-independent way. CpG DNA enabled the type I IFN-treated pDC to express IFN-α in the presence of NF-κB/p38 MAPK inhibitor, and chloroquine abrogated this effect. With CpG DNA, IRF-7, both constitutively and newly expressed, moved to the nuclei independently of NF-κB/p38 MAPK. These findings suggest that, in CpG DNA-stimulated human pDC, the induction of IRF-7, CXCL10, and CCL3 is mediated by the NF-κB/p38 MAPK pathway, and that IRF-7 is activated upstream of the activation of NF-κB/p38 MAPK in chloroquine-sensitive regulatory machinery, thereby leading to the expression of IFN-α.

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“…It is well known that bacterial DNA and synthetic oligodeoxynucleotides (ODNs) containing specific motifs centered on a CpG dinucleotide (CpG ODNs) are potent immunostimulatory agents (1)(2)(3)(4)(5). Unmethylated CpG sequences are present at a frequency of 1 in every 16 dinucleotides in bacteria, while mammals have a CG dinucleotide suppression with only one-fourth of the expected frequency, and the majority of the CpG sequences are methylated in mammals (6).…”

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confidence: 99%

Oligodeoxynucleotides containing CpG motifs can induce T cell–dependent arthritis in rats

Svelander¹,

Harris²,

Lorentzen³

et al. 2004

Arthritis & Rheumatism

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Objective. To investigate whether CpG oligodeoxynucleotides (ODNs) can induce or accelerate arthritis in rats.Methods. The CpG-induced response was studied by recording joint inflammation, cell activation in draining lymph nodes, and levels of the acute-phase reactant Conclusion. We demonstrated that injection with immunostimulatory CpG, both in phosphorothioatemodified and native forms, can induce a T celldependent joint-specific inflammation in LEW and LEW.1AV1 rat strains. This arthritis is preceded by signs of activation of the innate immune system. Since unmethylated CG dinucleotides are common in bacterial DNA but rare in mammalian DNA, our results indicate that exposure to bacterial DNA during infection may contribute to arthritis induction by amplifying the innate immune response.

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Oligonucleotide Sequences Required for Natural Killer Cell Activation

Cited by 117 publications

References 14 publications

Sequence-specific antitumor activity of a phosphorothioate oligodeoxyribonucleotide targeted to human C- raf kinase supports an antisense mechanism of action in vivo

Sequence-specific antitumor activity of a phosphorothioate oligodeoxyribonucleotide targeted to human C- raf kinase supports an antisense mechanism of action in vivo

Collaborative Action of NF-κB and p38 MAPK Is Involved in CpG DNA-Induced IFN-α and Chemokine Production in Human Plasmacytoid Dendritic Cells

Oligodeoxynucleotides containing CpG motifs can induce T cell–dependent arthritis in rats

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