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PMID: 12384593 Published · ppublish English Journal Article

'Immunomers'--novel 3'-3'-linked CpG oligodeoxyribonucleotides as potent immunomodulatory agents.

Nucleic acids research ·Vol. 30 ·No. 20 ·2002-10-15 ·Pages 4460-9

Yu D, Kandimalla ER, Bhagat L, Tang JY, Cong Y, Tang J, Agrawal S

Abstract

Oligodeoxyribonucleotides containing CpG dinucleotides (CpG DNAs) are currently being evaluated as novel immunomodulators in clinical trials. Recently, we showed that an accessible 5' end is required for immunostimulatory activity and blocking the 5' end of CpG DNA by conjugation of certain ligands abrogates immunostimulatory activity. Based on these results, we designed and synthesized 3'-3'-linked CpG DNAs that contained two or more identical CpG DNA segments, referred to here as 'immunomers'. The use of solid support bearing diDMT-glyceryl-linker permitted convenient synthesis of immunomers with both segments synthesized simultaneously, giving better yields and purity. The in vitro and in vivo studies suggest that as a result of accessibility to two 5' ends for recognition, immunomers show an enhanced immunostimulatory activity compared with linear CpG DNAs. We also studied the suitability of a number of different linkers for attaching the two segments of immunomers. A C3-linker was found to be optimal for joining the two segments of immunomers. Incorporation of multiple linkers between the two segments of immunomers resulted in different cytokine profiles depending on the nature and number of linkers incorporated. Additionally, the length of immunomer also plays a significant role in inducing immune responses. An immunomer containing 11 nt in each segment showed the highest activity and an 11mer linear CpG DNA failed to stimulate an immune response. These results suggest that immunomers have several advantages over conventional linear CpG DNAs for immunomodulatory activity studies.

MeSH Terms
Adjuvants, Immunologic/chemical synthesis,chemistry,pharmacology Animals Cell Line Cells, Cultured Dose-Response Relationship, Drug Female Interleukins/biosynthesis Mice Mice, Inbred BALB C NF-kappa B/metabolism Oligodeoxyribonucleotides/chemical synthesis,chemistry,pharmacology
Chemicals
Adjuvants, Immunologic CPG-oligonucleotide Interleukins NF-kappa B Oligodeoxyribonucleotides
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Yu Dong
Hybridon, Inc., 345 Vassar Street, Cambridge, MA 02139, USA.
Kandimalla Ekambar R
Bhagat Lakshmi
Tang Jin-Yan
Cong Yanping
Tang Jimmy
Agrawal Sudhir
References (52)
52 references, click to expand
  1. Human peripheral blood cells differentially recognize and respond to two distinct CPG motifs.
    J Immunol. 2001 Feb 15;166(4):2372-7 PMID: 11160295
  2. Activation of human B cells by phosphorothioate oligodeoxynucleotides.
    J Clin Invest. 1996 Sep 1;98(5):1119-29 PMID: 8787674
  3. Identification of CpG oligonucleotide sequences with high induction of IFN-alpha/beta in plasmacytoid dendritic cells.
    Eur J Immunol. 2001 Jul;31(7):2154-63 PMID: 11449369
  4. Antimetastatic effect of CpG DNA mediated by type I IFN.
    Cancer Res. 2001 Jul 15;61(14):5523-8 PMID: 11454702
  5. Modulation of immunostimulatory activity of CpG oligonucleotides by site-specific deletion of nucleobases.
    Bioorg Med Chem Lett. 2001 Sep 3;11(17):2263-7 PMID: 11527711
  6. Nucleic acid for the treatment of cancer: genetic vaccines and DNA adjuvants.
    Curr Pharm Des. 2001 Nov;7(16):1641-67 PMID: 11562304
  7. Recent developments in adjuvants for vaccines against infectious diseases.
    Biomol Eng. 2001 Oct 15;18(3):69-85 PMID: 11566599
  8. DNA immunomodulation of asthma.
    Clin Allergy Immunol. 2002;16:551-64 PMID: 11577559
  9. Immunostimulatory activity of CpG oligonucleotides containing non-ionic methylphosphonate linkages.
    Bioorg Med Chem. 2001 Nov;9(11):2803-8 PMID: 11597460
  10. Fighting infection using immunomodulatory agents.
    Expert Opin Biol Ther. 2001 Jul;1(4):641-53 PMID: 11727500
  11. Interactions between bacterial CpG-DNA and TLR9 bridge innate and adaptive immunity.
    Curr Opin Microbiol. 2002 Feb;5(1):62-9 PMID: 11834371
  12. Synergism between cytosine-guanine oligodeoxynucleotides and monoclonal antibody in the treatment of lymphoma.
    Semin Oncol. 2002 Feb;29(1 Suppl 2):93-7 PMID: 11842395
  13. CpG motifs in bacterial DNA and their immune effects.
    Annu Rev Immunol. 2002;20:709-60 PMID: 11861616
  14. Toll-like receptor 9 mediates CpG-DNA signaling.
    J Leukoc Biol. 2002 Mar;71(3):538-44 PMID: 11867692
  15. Medicinal chemistry and therapeutic potential of CpG DNA.
    Trends Mol Med. 2002 Mar;8(3):114-21 PMID: 11879771
  16. DNA-based approaches to the treatment of allergies.
    Curr Opin Mol Ther. 2002 Feb;4(1):64-71 PMID: 11883696
  17. Immunostimulatory properties of phosphorothioate CpG DNA containing both 3'-5'- and 2'-5'-internucleotide linkages.
    Nucleic Acids Res. 2002 Apr 1;30(7):1613-9 PMID: 11917022
  18. Towards optimal design of second-generation immunomodulatory oligonucleotides.
    Curr Opin Mol Ther. 2002 Apr;4(2):122-9 PMID: 12044033
  19. Potent CpG oligonucleotides containing phosphodiester linkages: in vitro and in vivo immunostimulatory properties.
    Biochem Biophys Res Commun. 2002 Sep 13;297(1):83-90 PMID: 12220512
  20. Conjugation of ligands at the 5'-end of CpG DNA affects immunostimulatory activity.
    Bioconjug Chem. 2002 Sep-Oct;13(5):966-74 PMID: 12236778
  21. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.
    Anal Biochem. 1976 May 7;72:248-54 PMID: 942051
  22. Antitumor activity of deoxyribonucleic acid fraction from Mycobacterium bovis BCG. I. Isolation, physicochemical characterization, and antitumor activity.
    J Natl Cancer Inst. 1984 Apr;72(4):955-62 PMID: 6200641
  23. In vitro augmentation of natural killer cell activity and production of interferon-alpha/beta and -gamma with deoxyribonucleic acid fraction from Mycobacterium bovis BCG.
    Jpn J Cancer Res. 1988 Jul;79(7):866-73 PMID: 2459094
  24. Rapid detection of octamer binding proteins with 'mini-extracts', prepared from a small number of cells.
    Nucleic Acids Res. 1989 Aug 11;17(15):6419 PMID: 2771659
  25. Absorbance melting curves of RNA.
    Methods Enzymol. 1989;180:304-25 PMID: 2482421
  26. Pharmacokinetics, biodistribution, and stability of oligodeoxynucleotide phosphorothioates in mice.
    Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7595-9 PMID: 1881900
  27. Stimulation of in vitro murine lymphocyte proliferation by bacterial DNA.
    J Immunol. 1991 Sep 15;147(6):1759-64 PMID: 1890302
  28. Synthetic oligonucleotides with particular base sequences from the cDNA encoding proteins of Mycobacterium bovis BCG induce interferons and activate natural killer cells.
    Microbiol Immunol. 1992;36(1):55-66 PMID: 1584073
  29. Unique palindromic sequences in synthetic oligonucleotides are required to induce IFN [correction of INF] and augment IFN-mediated [correction of INF] natural killer activity.
    J Immunol. 1992 Jun 15;148(12):4072-6 PMID: 1376349
  30. Immune stimulation by an antisense oligomer complementary to the rev gene of HIV-1.
    Biochem Pharmacol. 1993 May 25;45(10):2037-43 PMID: 8512586
  31. Capped oligodeoxynucleotide phosphorothioates. Pharmacokinetics and stability in mice.
    Ann N Y Acad Sci. 1992 Oct 28;660:318-20 PMID: 1340145
  32. Pharmacokinetics, biodistribution, and stability of capped oligodeoxynucleotide phosphorothioates in mice.
    Antisense Res Dev. 1993 Fall;3(3):277-84 PMID: 8286928
  33. CpG motifs in bacterial DNA trigger direct B-cell activation.
    Nature. 1995 Apr 6;374(6522):546-9 PMID: 7700380
  34. Properties of exonuclease-resistant, psoralen-conjugated oligodeoxyribonucleotides in vitro and in cell culture.
    Antisense Res Dev. 1994 Winter;4(4):231-41 PMID: 7734938
  35. Bacterial DNA induces murine interferon-gamma production by stimulation of interleukin-12 and tumor necrosis factor-alpha.
    Cell Immunol. 1996 Jan 10;167(1):72-8 PMID: 8548847
  36. CpG motifs present in bacteria DNA rapidly induce lymphocytes to secrete interleukin 6, interleukin 12, and interferon gamma.
    Proc Natl Acad Sci U S A. 1996 Apr 2;93(7):2879-83 PMID: 8610135
  37. Immunostimulatory DNA sequences necessary for effective intradermal gene immunization.
    Science. 1996 Jul 19;273(5273):352-4 PMID: 8662521
  38. Macrophages ingest and are activated by bacterial DNA.
    J Immunol. 1996 Sep 1;157(5):2116-22 PMID: 8757335
  39. In vivo metabolic profile of a phosphorothioate oligodeoxyribonucleotide.
    Antisense Nucleic Acid Drug Dev. 1997 Jun;7(3):159-65 PMID: 9212906
  40. Pattern and kinetics of cytokine production following administration of phosphorothioate oligonucleotides in mice.
    Antisense Nucleic Acid Drug Dev. 1997 Oct;7(5):495-502 PMID: 9361908
  41. Oligodeoxynucleotides containing palindrome sequences with internal 5'-CpG-3' act directly on human NK and activated T cells to induce IFN-gamma production in vitro.
    J Immunol. 1999 Oct 1;163(7):3642-52 PMID: 10490958
  42. Evaluation of different types of end-capping modifications on the stability of oligonucleotides toward 3'- and 5'-exonucleases.
    Nucleosides Nucleotides. 1999 Sep;18(9):2051-69 PMID: 10549151
  43. Site of chemical modifications in CpG containing phosphorothioate oligodeoxynucleotide modulates its immunostimulatory activity.
    Bioorg Med Chem Lett. 1999 Dec 20;9(24):3453-8 PMID: 10617090
  44. Delineation of a CpG phosphorothioate oligodeoxynucleotide for activating primate immune responses in vitro and in vivo.
    J Immunol. 2000 Feb 1;164(3):1617-24 PMID: 10640783
  45. DNA vaccines: immunology, application, and optimization*.
    Annu Rev Immunol. 2000;18:927-74 PMID: 10837079
  46. Immunostimulatory activity of CpG containing phosphorothioate oligodeoxynucleotide is modulated by modification of a single deoxynucleoside.
    Bioorg Med Chem Lett. 2000 May 15;10(10):1051-4 PMID: 10843214
  47. A Toll-like receptor recognizes bacterial DNA.
    Nature. 2000 Dec 7;408(6813):740-5 PMID: 11130078
  48. Accessible 5'-end of CpG-containing phosphorothioate oligodeoxynucleotides is essential for immunostimulatory activity.
    Bioorg Med Chem Lett. 2000 Dec 4;10(23):2585-8 PMID: 11128629
  49. Non-specific antiviral activity of antisense molecules targeted to the E1 region of human papillomavirus.
    Antiviral Res. 2000 Dec;48(3):187-96 PMID: 11164505
  50. Sequence identity of the n-1 product of a synthetic oligonucleotide.
    Nucleic Acids Res. 1995 Jun 11;23(11):1841-4 PMID: 7596808
  51. Effect of different chemically modified oligodeoxynucleotides on immune stimulation.
    Biochem Pharmacol. 1996 Jan 26;51(2):173-82 PMID: 8615886
  52. Effect of chemical modifications of cytosine and guanine in a CpG-motif of oligonucleotides: structure-immunostimulatory activity relationships.
    Bioorg Med Chem. 2001 Mar;9(3):807-13 PMID: 11310616
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/gkf582
Published
2002-10-15
Pages
4460-9
Language
English
Region
England
NLM ID
0411011
PMCID
PMC137145
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/gkf582 PMC Full Text View journal details
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