Abstract
A specific set of immune functions is switched on in response to DNA containing unmethylated CpG dinucleotides in particular base contexts (‘CpG motifs’). Plasmids, viral vectors and antisense oligodeoxynucleotides used for DNA vaccination, gene replacement or gene blockade contain immunostimulatory CpG motifs which may have independent biological activity. Although the immune stimulatory effects of CpG motifs on murine cells are well established, the evaluation of their possible effects on human cells is complicated by the higher LPS sensitivity of human leukocytes compared with those in mice. To address this issue, we analyzed CpG- and LPS-mediated immune activation of human PBMC. The biologic activity of LPS could be detected within 4 h using intracellular TNF staining of monocytes with flow cytometry at concentrations just one-twentieth (0.0014 EU/ml) of the lower detection limit for the routinely used LAL assay (0.03 EU/ml). In contrast to the rapid LPS response, CpG DNA-stimulated TNF and IL-6 synthesis in human mono- cytes was not detectable until 18 h. E. coli DNA induced IL-6 synthesis in a concentration-dependent manner (30 μg/ml E. coli DNA: 409 pg/ml ± 75 pg/ml, n = 7, IL-6 ELISA), but calf thymus DNA did not (<10 pg/ml). likewise, the cpg oligodeoxynucleotides 1760 (phospho- rothioate) and 2059 (unmodified) induced il-6 synthesis, but the corresponding control oligonucleotides 1908 and 2077 did not. cpg dna and lps enhanced il-6 synthesis synergistically. icam-1-expression of monocytes was increased 4.6-fold by e. coli dna, 3.5-fold by 1760 and three-fold by 2059, compared with 3.6-fold by a maximal lps stimulus and no change with non-cpg dna. in conclusion, cpg-motifs induce tnf, il-6 and icam-1 expression in human monocytes, but the kinetics of this differ from that induced by lps, which makes it possible to distinguish immune activation by these agents. these results have important implications for the clinical development of therapeutic dna in humans.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Davis HL . Plasmid DNA expression systems for the purpose of immunization Curr Opin Biotechnol 1997 8: 635–646
Hartmann G et al. Specific suppression of human tumor necrosis factor-alpha synthesis by antisense oligodeoxynucleotides Antisense Nucleic Acid Drug Dev 1996 6: 291–299
Branch AD . A good antisense molecule is hard to find Trends Biochem Sci 1998 23: 45–50
Agrawal S, Iyer RP . Perspectives in antisense therapeutics Pharmacol Ther 1997 76: 151–160
Krieg AM et al. CpG motifs in bacterial DNA trigger direct B-cell activation Nature 1995 374: 546–549
Krieg AM, Yi AK, Schorr J, Davis HL . The role of CpG dinucleotides in DNA vaccines Trends Microbiol 1998 6: 23–27
Krieg AM . An innate immune defense mechanism based on the recognition of CpG motifs in microbial DNA J Lab Clin Med 1996 128: 128–133
Krieg AM . Lymphocyte activation by CpG dinucleotide motifs in prokaryotic DNA Trends Microbiol 1996 4: 73–76
Sato Y et al. Immunostimulatory DNA sequences necessary for effective intradermal gene immunization Science 1996 273: 352–354
Klinman DM, Yamshchikov G, Ishigatsubo Y . Contribution of CpG motifs to the immunogenicity of DNA vaccines J Immunol 1997 158: 3635–3639
Hartmann G et al. Cytokines and therapeutic oligonucleotides Cytok Cell Molec Ther 1997 3: 247–256
Krieg A . How to exclude immunostimulatory effects of antisense oligonucleotides. In: Hartmann G, Endres S (eds). Manual of Antisense Methodology Kluwer Academic: Boston 1999 (in press
Moldoveanu Z, Love-Homan L, Huang WQ, Krieg AM . CpG DNA, a novel immune enhancer for systemic and mucosal immunization with influenza virus Vaccine 1998 16: 1216–1224
Davis HL et al. CpG DNA is a potent enhancer of specific immunity in mice immunized with recombinant hepatitis B surface antigen J Immunol 1998 160: 870–876
Weiner GJ et al. Immunostimulatory oligodeoxynucleotides containing the CpG motif are effective as immune adjuvants in tumor antigen immunization Proc Natl Acad Sci USA 1997 94: 10833–10837
Wooldridge JE, Ballas Z, Krieg AM, Weiner GJ . Immunostimulatory oligodeoxynucleotides containing CpG motifs enhance the efficacy of monoclonal antibody therapy of lymphoma Blood 1997 89: 2994–2998
Roman M et al. Immunostimulatory DNA sequences function as T helper-1-promoting adjuvants Nature Med 1997 3: 849–854
Chu RS et al. CpG oligodeoxynucleotides act as adjuvants that switch on T helper 1 (Th1) immunity J Exp Med 1997 186: 1623–1631
Kline JN et al. Modulation of airway inflammation by CpG oligodeoxynucleotides in a murine model of asthma J Immunol 1998 160: 2555–2559
Krieg AM, Love-Homan L, Yi AK, Harty JT . CpG DNA induces sustained IL-12 expression in vivo and resistance to Listeria monocytogenes challenge J Immunol 1998 161: 2428–2434
Zimmermann S et al. CpG oligodeoxynucleotides trigger protective and curative Th1 responses in lethal murine leishmaniasis J Immunol 1998 160: 3627–3630
Freund J, Casals J, Hosmer EP . Sensitization and antibody formation after injection of tubercle bacilli and paraffin oil Proc Soc Exp Biol Med 1937 37: 509–513
Freund J, McDermot K . Sensitization to horse serum by means of adjuvants Proc Soc Exp Bio NY 1942 49: 548–553
Herbert WJ . Mineral-oil adjuvants and the immunization of laboratory animals. In: Weir DM (ed) . Handbook of Experimental Immunology Blackwell Scientific: Oxford 1978 179–193
Yamamoto S et al. DNA from bacteria, but not from vertebrates, induces interferons, activates natural killer cells and inhibits tumor growth Microbiol Immunol 1992 36: 983–997
Messina JP, Gilkeson GS, Pisetsky DS . Stimulation of in vitro murine lymphocyte proliferation by bacterial DNA J Immunol 1991 147: 1759–1764
Munoz J . Effect of bacteria and bacterial products on antibody response Adv Immunol 1964 4: 397–440
Stacey KJ, Sweet MJ, Hume DA . Macrophages ingest and are activated by bacterial DNA J Immunol 1996 157: 2116–2122
Sweet MJ et al. IFN-gamma primes macrophage responses to bacterial DNA J Interferon Cytok Res 1998 18: 263–271
Lamont AG, Adorini L . IL-12: a key cytokine in immune regulation Immunol Today 1996 17: 214–217
Eigler A, Sinha B, Hartmann G, Endres S . Taming TNF: strategies to restrain this proinflammatory cytokine Immunol Today 1997 18: 487–492
Chace JH et al. Bacterial DNA-induced NK cell IFN-gamma production is dependent on macrophage secretion of IL-12 Clin Immunol Immunopathol 1997 84: 185–193
Cowdery JS, Chace JH, Yi AK, Krieg AM . Bacterial DNA induces NK cells to produce IFN-gamma in vivo and increases the toxicity of lipopolysaccharides J Immunol 1996 156: 4570–4575
Ballas ZK, Rasmussen WL, Krieg AM . Induction of NK activity in murine and human cells by CpG motifs in oligodeoxynucleotides and bacterial DNA J Immunol 1996 157: 1840–1845
Yi AK, Chace JH, Cowdery JS, Krieg AM . IFN-gamma promotes IL-6 and IgM secretion in response to CpG motifs in bacterial DNA and oligodeoxynucleotides J Immunol 1996 156: 558–564
Yi AK et al. Rapid immune activation by CpG motifs in bacterial DNA. Systemic induction of IL-6 transcription through an antioxidant-sensitive pathway J Immunol 1996 157: 5394–5402
Klinman DM et al. CpG motifs present in bacteria DNA rapidly induce lymphocytes to secrete interleukin 6, interleukin 12, and interferon gamma Proc Natl Acad Sci USA 1996 93: 2879–2883
Macfarlane DE, Manzel L . Antagonism of immunostimulatory CpG-oligodeoxynucleotides by quinacrine, chloroquine, and structurally related compounds J Immunol 1998 160: 1122–1131
Yamamoto T et al. Synthetic oligonucleotides with certain palindromes stimulate interferon production of human peripheral blood lymphocytes in vitro Jpn J Cancer Res 1994 85: 775–779
Liang H et al. Activation of human B cells by phosphorothioate oligodeoxynucleotides J Clin Invest 1996 98: 1119–1129
Branda RF et al. B-cell proliferation and differentiation in common variable immunodeficiency patients produced by an antisense oligomer to the rev gene of HIV-1 Clin Immunol Immunopathol 1996 79: 115–121
Branda RF et al. Amplification of antibody production by phosphorothioate oligodeoxynucleotides J Lab Clin Med 1996 128: 329–338
Yi AK et al. CpG motifs in bacterial DNA activate leukocytes through the pH-dependent generation of reactive oxygen species J Immunol 1998 160: 4755–4761
Gao JX et al. Generation of dendritic cell-like antigen-presenting cells in long-term mixed leucocyte culture: phenotypic and functional studies Immunology 1997 91: 135–144
Yacyshyn BR et al. A placebo-controlled trial of ICAM-1 antisense oligonucleotide in the treatment of Crohn’s disease Gastroenterology 1998 114: 1133–1142
Ohta Y et al. Direct antigen presentation through binding of donor intercellular adhesion molecule-1 to recipient lymphocyte function-associated antigen-1 molecules in xenograft rejection Transplantation 1998 65: 1094–1100
Taguchi M et al. Patterns for RANTES secretion and intercellular adhesion molecule 1 expression mediate transepithelial T cell traffic based on analyses in vitro and in vivo J Exp Med 1998 187: 1927–1940
Sparwasser T et al. Macrophages sense pathogens via DNA motifs: induction of tumor necrosis factor-alpha-mediated shock Eur J Immunol 1997 27: 1671–1679
Schwartz DA et al. CpG motifs in bacterial DNA cause inflammation in the lower respiratory tract J Clin Invest 1997 100: 68–73
Sartor RB . Review article: how relevant to human inflammatory bowel disease are current animal models of intestinal inflammation? Aliment Pharmacol Ther 1997 3: 89–96
Elson CO, Sartor RB, Tennyson GS, Riddel RH . Experimental models of inflammatory bowel disease Gastroenterology 1995 109: 1344–1367
Sun S, Kishimoto H, Sprent J . DNA as an adjuvant: capacity of insect DNA and synthetic oligodeoxynucleotides to augment T cell responses to specific antigen J Exp Med 1998 187: 1145–1150
Hartmann G, Krug A, Waller-Fontaine K, Endres S . Oligodeoxynucleotides enhance lipopolysaccharide-stimulated synthesis of tumor necrosis factor: dependence on phosphorothioate modification and reversal by heparin Mol Med 1996 2: 429–438
Hartmann G et al. Spontaneous and cationic lipid-mediated uptake of antisense oligonucleotides in human monocytes and lymphocytes J Pharmacol Exp Ther 1998 285: 920–928
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hartmann, G., Krieg, A. CpG DNA and LPS induce distinct patterns of activation in human monocytes. Gene Ther 6, 893–903 (1999). https://doi.org/10.1038/sj.gt.3300880
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.gt.3300880
Keywords
This article is cited by
-
Multi-step screening of DNA/lipid nanoparticles and co-delivery with siRNA to enhance and prolong gene expression
Nature Communications (2022)
-
Cleavable conjugation of CpG oligodeoxynucleotides onto microparticles for facile release and cytokine induction in macrophages
Applied Biological Chemistry (2017)
-
Expansion and preferential activation of the CD14+CD16+ monocyte subset during multiple sclerosis
Immunology & Cell Biology (2014)
-
R-848 triggers the expression of TLR7/8 and suppresses HIV replication in monocytes
BMC Infectious Diseases (2012)
-
Listeria-derived ActA is an effective adjuvant for primary and metastatic tumor immunotherapy
Cancer Immunology, Immunotherapy (2010)