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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 a gene cluster in PBMC. A gene cluster containing 77 genes was up-regulated more than two-fold in PBMC after stimulation with CpG ODN. Data were analyzed by applying a hierarchical-tree algorithm to the normalized intensity. The color code for the signal strength in the classification scheme is shown in the box on the left. Induced genes are indicated by shades of red; repressed genes are indicated by shades of blue. Exactly the same experiments were performed with PBMCs from three individual donors to confirm the reproducibility of the present experiments. All of these genes were consistently included in a CpG-inducible cluster in the same fashion even though cRNA elongation was not completed in a single experiment. * GenBank accession number
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Figure 1: CpG ODN mediated up-regulation of a gene cluster in PBMC. A gene cluster containing 77 genes was up-regulated more than two-fold in PBMC after stimulation with CpG ODN. Data were analyzed by applying a hierarchical-tree algorithm to the normalized intensity. The color code for the signal strength in the classification scheme is shown in the box on the left. Induced genes are indicated by shades of red; repressed genes are indicated by shades of blue. Exactly the same experiments were performed with PBMCs from three individual donors to confirm the reproducibility of the present experiments. All of these genes were consistently included in a CpG-inducible cluster in the same fashion even though cRNA elongation was not completed in a single experiment. * GenBank accession number

Mentions: Using hierarchical clustering-analysis of the gene expression profiles of approximately 10,000 genes, we identified a cluster containing 77 genes which were up-regulated by CpG ODN (Fig. 1). This cluster was further sub-divided into two clusters on the basis of time-kinetics. Early-response genes, including IL-6, lymphotoxin α (LTA) and granulocyte-macrophage colony-stimulating factor (GM-CSF, CSF2), were up-regulated predominantly 3 to 6 hours after stimulation with CpG ODN. In contrast, late-response genes, including IFN-induced protein with tetratricopeptide repeats 1 (IFIT1), IFIT2, 2',5'-oligoadenylate synthetase 1 (OAS1), OAS2, myxovirus (influenza virus) resistance 1 (Mx1), Mx2 and IFN-γ inducible protein 10 (IP-10), were up-regulated predominantly 6 to 24 hours after stimulation with CpG ODN. Some CpG ODN-inducible genes, especially such late-response genes as IFIT1, OAS1 and IP-10, have been shown to be regulated by type I IFN and/or type II IFNs [18]. However, mRNAs for all type I IFNs were judged to be 'Absent' by Microarray Suite software, which means that the mRNAs are below the detection limit of the GeneChip® system (data not shown).


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 a gene cluster in PBMC. A gene cluster containing 77 genes was up-regulated more than two-fold in PBMC after stimulation with CpG ODN. Data were analyzed by applying a hierarchical-tree algorithm to the normalized intensity. The color code for the signal strength in the classification scheme is shown in the box on the left. Induced genes are indicated by shades of red; repressed genes are indicated by shades of blue. Exactly the same experiments were performed with PBMCs from three individual donors to confirm the reproducibility of the present experiments. All of these genes were consistently included in a CpG-inducible cluster in the same fashion even though cRNA elongation was not completed in a single experiment. * GenBank accession number
© Copyright Policy
Related In: Results  -  Collection

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

Figure 1: CpG ODN mediated up-regulation of a gene cluster in PBMC. A gene cluster containing 77 genes was up-regulated more than two-fold in PBMC after stimulation with CpG ODN. Data were analyzed by applying a hierarchical-tree algorithm to the normalized intensity. The color code for the signal strength in the classification scheme is shown in the box on the left. Induced genes are indicated by shades of red; repressed genes are indicated by shades of blue. Exactly the same experiments were performed with PBMCs from three individual donors to confirm the reproducibility of the present experiments. All of these genes were consistently included in a CpG-inducible cluster in the same fashion even though cRNA elongation was not completed in a single experiment. * GenBank accession number
Mentions: Using hierarchical clustering-analysis of the gene expression profiles of approximately 10,000 genes, we identified a cluster containing 77 genes which were up-regulated by CpG ODN (Fig. 1). This cluster was further sub-divided into two clusters on the basis of time-kinetics. Early-response genes, including IL-6, lymphotoxin α (LTA) and granulocyte-macrophage colony-stimulating factor (GM-CSF, CSF2), were up-regulated predominantly 3 to 6 hours after stimulation with CpG ODN. In contrast, late-response genes, including IFN-induced protein with tetratricopeptide repeats 1 (IFIT1), IFIT2, 2',5'-oligoadenylate synthetase 1 (OAS1), OAS2, myxovirus (influenza virus) resistance 1 (Mx1), Mx2 and IFN-γ inducible protein 10 (IP-10), were up-regulated predominantly 6 to 24 hours after stimulation with CpG ODN. Some CpG ODN-inducible genes, especially such late-response genes as IFIT1, OAS1 and IP-10, have been shown to be regulated by type I IFN and/or type II IFNs [18]. However, mRNAs for all type I IFNs were judged to be 'Absent' by Microarray Suite software, which means that the mRNAs are below the detection limit of the GeneChip® system (data not shown).

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