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Interferon-alpha/beta receptor-mediated selective induction of a gene cluster by CpG oligodeoxynucleotide 2006.

Kato A, Homma T, Batchelor J, Hashimoto N, Imai S, Wakiguchi H, Saito H, Matsumoto K - BMC Immunol. (2003)

Bottom Line: Oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODN) are known to exert a strong adjuvant effect on Th1 immune responses.Blocking with mAb against IFN-alpha/beta receptor strongly inhibited the induction of these IFN-inducible genes by CpG ODN.This study provides new information regarding the possible immunomodulatory effects of CpG ODN in vivo via an IFN-alpha/beta receptor-mediated paracrine pathway.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Allergy & Immunology, National Research Institute for Child Health & Development, Tokyo, Japan. atkato@nch.go.jp

ABSTRACT

Background: Oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODN) are known to exert a strong adjuvant effect on Th1 immune responses. Although several genes have been reported, no comprehensive study of the gene expression profiles in human cells after stimulation with CpG ODN has been reported.

Results: This study was designed to identify a CpG-inducible gene cluster that potentially predicts for the molecular mechanisms of clinical efficacy of CpG ODN, by determining mRNA expression in human PBMC after stimulation with CpG ODN. PBMCs were obtained from the peripheral blood of healthy volunteers and cultured in the presence or absence of CpG ODN 2006 for up to 24 hours. The mRNA expression profile was evaluated using a high-density oligonucleotide probe array, GeneChip. Using hierarchical clustering-analysis, out of a total of 10,000 genes we identified a cluster containing 77 genes as having been up-regulated by CpG ODN. This cluster was further divided into two sub-clusters by means of time-kinetics. (1) Inflammatory cytokines such as IL-6 and GM-CSF were up-regulated predominantly 3 to 6 hours after stimulation with CpG ODN, presumably through activation of a transcription factor, NF-kappaB. (2) Interferon (IFN)-inducible anti-viral proteins, including IFIT1, OAS1 and Mx1, and Th1 chemoattractant IP-10, were up-regulated predominantly 6 to 24 hours after stimulation. Blocking with mAb against IFN-alpha/beta receptor strongly inhibited the induction of these IFN-inducible genes by CpG ODN.

Conclusion: This study provides new information regarding the possible immunomodulatory effects of CpG ODN in vivo via an IFN-alpha/beta receptor-mediated paracrine pathway.

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Related in: MedlinePlus

CpG ODN mediated up-regulation of mRNA for IL-6, LTA, OAS1, IFIT1, IP-10 and IFN-α2. PBMC were incubated in the presence or absence of 2 μg/ml CpG ODN 2006 for 6 hours. The mRNA levels of IL-6 (A), LTA (B), OAS1 (C), IFIT1 (D), IP-10 (E) and IFN-α2 (F) were determined by SYBR® green-based real-time quantitative RT-PCR. The results are shown as the mean ± SEM from four independent experiments. The copy number is expressed as the number of transcripts/ng of total RNA.
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Figure 2: CpG ODN mediated up-regulation of mRNA for IL-6, LTA, OAS1, IFIT1, IP-10 and IFN-α2. PBMC were incubated in the presence or absence of 2 μg/ml CpG ODN 2006 for 6 hours. The mRNA levels of IL-6 (A), LTA (B), OAS1 (C), IFIT1 (D), IP-10 (E) and IFN-α2 (F) were determined by SYBR® green-based real-time quantitative RT-PCR. The results are shown as the mean ± SEM from four independent experiments. The copy number is expressed as the number of transcripts/ng of total RNA.

Mentions: Up-regulation of the mRNA levels of five genes, IL-6, LTA, OAS1, IFIT1 and IP-10, was further confirmed by real-time PCR (Fig. 2). The magnitude of enhancement of the mRNA for IL-6, LTA, OAS1, IFIT1 and IP-10 by CpG ODN was 7-fold, 5-fold, 14-fold, 289-fold and 18-fold, respectively. However, induction of inflammatory genes such as IL-6 and GM-CSF by CpG ODN is very weak compared with LPS (Fig. 3). Up-regulation of the mRNA levels of CpG ODN-inducible genes, such as IP-10 and IL-6, was scarcely induced by GpC control ODN, being less than 1:10 when compared to the enhancement by CpG ODN 2006 (data not shown). In addition, induction of a low level of IFN-α was detectable by real-time PCR (Fig. 2).


Interferon-alpha/beta receptor-mediated selective induction of a gene cluster by CpG oligodeoxynucleotide 2006.

Kato A, Homma T, Batchelor J, Hashimoto N, Imai S, Wakiguchi H, Saito H, Matsumoto K - BMC Immunol. (2003)

CpG ODN mediated up-regulation of mRNA for IL-6, LTA, OAS1, IFIT1, IP-10 and IFN-α2. PBMC were incubated in the presence or absence of 2 μg/ml CpG ODN 2006 for 6 hours. The mRNA levels of IL-6 (A), LTA (B), OAS1 (C), IFIT1 (D), IP-10 (E) and IFN-α2 (F) were determined by SYBR® green-based real-time quantitative RT-PCR. The results are shown as the mean ± SEM from four independent experiments. The copy number is expressed as the number of transcripts/ng of total RNA.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC183869&req=5

Figure 2: CpG ODN mediated up-regulation of mRNA for IL-6, LTA, OAS1, IFIT1, IP-10 and IFN-α2. PBMC were incubated in the presence or absence of 2 μg/ml CpG ODN 2006 for 6 hours. The mRNA levels of IL-6 (A), LTA (B), OAS1 (C), IFIT1 (D), IP-10 (E) and IFN-α2 (F) were determined by SYBR® green-based real-time quantitative RT-PCR. The results are shown as the mean ± SEM from four independent experiments. The copy number is expressed as the number of transcripts/ng of total RNA.
Mentions: Up-regulation of the mRNA levels of five genes, IL-6, LTA, OAS1, IFIT1 and IP-10, was further confirmed by real-time PCR (Fig. 2). The magnitude of enhancement of the mRNA for IL-6, LTA, OAS1, IFIT1 and IP-10 by CpG ODN was 7-fold, 5-fold, 14-fold, 289-fold and 18-fold, respectively. However, induction of inflammatory genes such as IL-6 and GM-CSF by CpG ODN is very weak compared with LPS (Fig. 3). Up-regulation of the mRNA levels of CpG ODN-inducible genes, such as IP-10 and IL-6, was scarcely induced by GpC control ODN, being less than 1:10 when compared to the enhancement by CpG ODN 2006 (data not shown). In addition, induction of a low level of IFN-α was detectable by real-time PCR (Fig. 2).

Bottom Line: Oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODN) are known to exert a strong adjuvant effect on Th1 immune responses.Blocking with mAb against IFN-alpha/beta receptor strongly inhibited the induction of these IFN-inducible genes by CpG ODN.This study provides new information regarding the possible immunomodulatory effects of CpG ODN in vivo via an IFN-alpha/beta receptor-mediated paracrine pathway.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Allergy & Immunology, National Research Institute for Child Health & Development, Tokyo, Japan. atkato@nch.go.jp

ABSTRACT

Background: Oligodeoxynucleotides containing unmethylated CpG motifs (CpG ODN) are known to exert a strong adjuvant effect on Th1 immune responses. Although several genes have been reported, no comprehensive study of the gene expression profiles in human cells after stimulation with CpG ODN has been reported.

Results: This study was designed to identify a CpG-inducible gene cluster that potentially predicts for the molecular mechanisms of clinical efficacy of CpG ODN, by determining mRNA expression in human PBMC after stimulation with CpG ODN. PBMCs were obtained from the peripheral blood of healthy volunteers and cultured in the presence or absence of CpG ODN 2006 for up to 24 hours. The mRNA expression profile was evaluated using a high-density oligonucleotide probe array, GeneChip. Using hierarchical clustering-analysis, out of a total of 10,000 genes we identified a cluster containing 77 genes as having been up-regulated by CpG ODN. This cluster was further divided into two sub-clusters by means of time-kinetics. (1) Inflammatory cytokines such as IL-6 and GM-CSF were up-regulated predominantly 3 to 6 hours after stimulation with CpG ODN, presumably through activation of a transcription factor, NF-kappaB. (2) Interferon (IFN)-inducible anti-viral proteins, including IFIT1, OAS1 and Mx1, and Th1 chemoattractant IP-10, were up-regulated predominantly 6 to 24 hours after stimulation. Blocking with mAb against IFN-alpha/beta receptor strongly inhibited the induction of these IFN-inducible genes by CpG ODN.

Conclusion: This study provides new information regarding the possible immunomodulatory effects of CpG ODN in vivo via an IFN-alpha/beta receptor-mediated paracrine pathway.

Show MeSH
Related in: MedlinePlus