Molecular Medicine

, Volume 17, Issue 9–10, pp 910–916 | Cite as

Effect of Oxygen Levels on the Physiology of Dendritic Cells: Implications for Adoptive Cell Therapy

  • Diahnn Futalan
  • Chien-Tze Huang
  • Ingo G. H. Schmidt-Wolf
  • Marie Larsson
  • Davorka Messmer
Research Article


Dendritic cell (DC)-based adoptive tumor immunotherapy approaches have shown promising results, but the incidence of tumor regression is low and there is an evident call for identifying culture conditions that produce DCs with a more potent Th1 potential. Routinely, DCs are differentiated in CO2 incubators under atmospheric oxygen conditions (21% O2), which differ from physiological oxygen levels of only 3–5% in tissue, where most DCs reside. We investigated whether differentiation and maturation of DCs under physiological oxygen levels could produce more potent T-cell stimulatory DCs for use in adoptive immunotherapy. We found that immature DCs differentiated under physiological oxygen levels showed a small but significant reduction in their endocytic capacity. The different oxygen levels did not influence their stimuli-induced upregulation of cluster of differentiation 54 (CD54), CD40, CD83, CD86, C-C chemokine receptor type 7 (CCR7), C-X-C chemokine receptor type 4 (CXCR4) and human leukocyte antigen (HLA)-DR or the secretion of interleukin (IL)-6, tumor necrosis factor (TNF)-α and IL-10 in response to lipopolysaccharide (LPS) or a cytokine cocktail. However, DCs differentiated under physiological oxygen level secreted higher levels of IL-12(p70) after exposure to LPS or CD40 ligand. Immature DCs differentiated at physiological oxygen levels caused increased T-cell proliferation, but no differences were observed for mature DCs with regard to T-cell activation. In conclusion, we show that although DCs generated under atmospheric or physiological oxygen conditions are mostly similar in function and phenotype, DCs differentiated under physiological oxygen secrete larger amounts of IL-12(p70). This result could have implications for the use of ex vivo-generated DCs for clinical studies, since DCs differentiated at physiological oxygen could induce increased Th1 responses in vivo.



This work was supported by the U.S. Army Medical Research and Materiel Command under agreement number W81XWH-07-1-0412 to D Messmer, the Swedish Research Council AI52731 and the Swedish International Development Cooperation Agency (SIDA and VINNMER [Vinnova]) to M Larsson. The authors thank Jessie F Fecteau and Dan Seible for the critical reading of the manuscript.


  1. 1.
    Steinman RM, Banchereau J. (2007) Taking dendritic cells into medicine. Nature. 449:419–26.CrossRefPubMedGoogle Scholar
  2. 2.
    Steinman RM. (2007) Dendritic cells: understanding immunogenicity. Eur. J. Immunol. 37 Suppl 1:S53–60.CrossRefPubMedGoogle Scholar
  3. 3.
    Steinman RM, Mellman I. (2004) Immunotherapy: bewitched, bothered, and bewildered no more. Science. 305:197–200.CrossRefPubMedGoogle Scholar
  4. 4.
    Dhodapkar KM, Banerjee D, Steinman RM. (2005) Harnessing the immune system against human glioma. Ann. N. Y. Acad. Sci. 1062:13–21.CrossRefPubMedGoogle Scholar
  5. 5.
    Schuler G, Schuler-Thurner B, Steinman RM. (2003) The use of dendritic cells in cancer immunotherapy. Curr. Opin. Immunol. 15:138–47.CrossRefPubMedGoogle Scholar
  6. 6.
    Gilboa E. (2004) The promise of cancer vaccines. Nat. Rev. Cancer. 4:401–11.CrossRefPubMedGoogle Scholar
  7. 7.
    Gilboa E. (2007) DC-based cancer vaccines. J. Clin. Invest. 117:1195–203.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Banchereau J, Palucka AK. (2005) Dendritic cells as therapeutic vaccines against cancer. Nat. Rev. Immunol. 5:296–306.CrossRefPubMedGoogle Scholar
  9. 9.
    Nestle FO, Farkas A, Conrad C. (2005) Dendritic-cell-based therapeutic vaccination against cancer. Curr. Opin. Immunol. 17:163–9.CrossRefPubMedGoogle Scholar
  10. 10.
    Steinman RM, Pope M. (2002) Exploiting dendritic cells to improve vaccine efficacy. J. Clin. Invest. 109:1519–26.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Palucka AK, et al. (2005) Boosting vaccinations with peptide-pulsed CD34+ progenitor-derived dendritic cells can expand long-lived melanoma peptide-specific CD8+ T cells in patients with metastatic melanoma. J. Immunother. 28:158–68.CrossRefPubMedGoogle Scholar
  12. 12.
    Dhodapkar MV, Steinman RM, Krasovsky J, Munz C, Bhardwaj N. (2001) Antigen-specific inhibition of effector T cell function in humans after injection of immature dendritic cells. J. Exp. Med. 193:233–8.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Palucka AK, et al. (2003) Single injection of CD34+ progenitor-derived dendritic cell vaccine can lead to induction of T-cell immunity in patients with stage IV melanoma. J. Immunother. 26:432–9.CrossRefPubMedGoogle Scholar
  14. 14.
    Dhodapkar MV, et al. (1999) Rapid generation of broad T-cell immunity in humans after a single injection of mature dendritic cells. J. Clin. Invest. 104:173–80.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Yu JS, et al. (2004) Vaccination with tumor lysatepulsed dendritic cells elicits antigen-specific, cytotoxic T-cells in patients with malignant glioma. Cancer Res. 64:4973–9.CrossRefPubMedGoogle Scholar
  16. 16.
    Lee WC, et al. (2005) Vaccination of advanced hepatocellular carcinoma patients with tumor lysate-pulsed dendritic cells: a clinical trial. J. Immunother. 28:496–504.CrossRefPubMedGoogle Scholar
  17. 17.
    Chang AE, et al. (2002) A phase I trial of tumor lysate-pulsed dendritic cells in the treatment of advanced cancer. Clin. Cancer Res. 8:1021–32.PubMedGoogle Scholar
  18. 18.
    Su Z, et al. (2005) Telomerase mRNA-transfected dendritic cells stimulate antigen-specific CD8+ and CD4+ T cell responses in patients with metastatic prostate cancer. J. Immunol. 174:3798–807.CrossRefPubMedGoogle Scholar
  19. 19.
    Van Tendeloo VF, et al. (2001) Highly efficient gene delivery by mRNA electroporation in human hematopoietic cells: superiority to lipofection and passive pulsing of mRNA and to electroporation of plasmid cDNA for tumor antigen loading of dendritic cells. Blood. 98:49–56.CrossRefPubMedGoogle Scholar
  20. 20.
    Nair SK, et al. (1998) Induction of primary carcinoembryonic antigen (CEA)-specific cytotoxic T lymphocytes in vitro using human dendritic cells transfected with RNA. Nat. Biotechnol. 16:364–9.CrossRefPubMedGoogle Scholar
  21. 21.
    Gilboa E, Vieweg J. (2004) Cancer immunotherapy with mRNA-transfected dendritic cells. Immunol. Rev. 199:251–63.CrossRefPubMedGoogle Scholar
  22. 22.
    Bubenik J. (2001) Genetically engineered dendritic cell-based cancer vaccines (Review). Int. J. Oncol. 18:475–8.PubMedGoogle Scholar
  23. 23.
    Steinman RM. (2003) The control of immunity and tolerance by dendritic cell. Pathol. Biol. (Paris). 51:59–60.CrossRefGoogle Scholar
  24. 24.
    Steinman RM, Hawiger D, Nussenzweig MC. (2003) Tolerogenic dendritic cells. Annu. Rev. Immunol. 21:685–711.CrossRefPubMedGoogle Scholar
  25. 25.
    De Vries IJ, et al. (2003) Effective migration of antigen-pulsed dendritic cells to lymph nodes in melanoma patients is determined by their maturation state. Cancer Res. 63:12–7.PubMedGoogle Scholar
  26. 26.
    Schuler-Thurner B, et al. (2002) Rapid induction of tumor-specific type 1 T helper cells in metastatic melanoma patients by vaccination with mature, cryopreserved, peptide-loaded monocyte-derived dendritic cells. J. Exp. Med. 195:1279–88.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Jonuleit H, et al. (1997) Pro-inflammatory cytokines and prostaglandins induce maturation of potent immunostimulatory dendritic cells under fetal calf serum-free conditions. Eur. J. Immunol. 27:3135–42.CrossRefPubMedGoogle Scholar
  28. 28.
    Palucka AK, et al. (2006) Dendritic cells loaded with killed allogeneic melanoma cells can induce objective clinical responses and MART-1 specific CD8+ T-cell immunity. J. Immunother. 29:545–57.CrossRefPubMedGoogle Scholar
  29. 29.
    O’Rourke MG, et al. (2003) Durable complete clinical responses in a phase I/II trial using an autologous melanoma cell/dendritic cell vaccine. Cancer Immunol. Immunother. 52:387–95.PubMedGoogle Scholar
  30. 30.
    Campbell JA. (1925) The influence of O(2)-tension in the inspired air upon the O(2)-tension in the tissues. J. Physiol. 60:20–9.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Laser H. (1937) Tissue metabolism under the influence of low oxygen tension. Biochem. J. 31:1671–6.CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Caldwell CC, et al. (2001) Differential effects of physiologically relevant hypoxic conditions on T lymphocyte development and effector functions. J. Immunol. 167:6140–9.CrossRefPubMedGoogle Scholar
  33. 33.
    Atkuri KR, Herzenberg LA, Niemi AK, Cowan T. (2007) Importance of culturing primary lymphocytes at physiological oxygen levels. Proc. Natl. Acad. Sci. U. S. A. 104:4547–52.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Atkuri KR, Herzenberg LA. (2005) Culturing at atmospheric oxygen levels impacts lymphocyte function. Proc. Natl. Acad. Sci. U. S. A. 102:3756–9.CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Yang M, et al. (2009) Hypoxia skews dendritic cells to a T helper type 2-stimulating phenotype and promotes tumour cell migration by dendritic cell-derived osteopontin. Immunology. 128:e237–49.CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Wang Q, et al. (2010) Reoxygenation of hypoxia-differentiated dentritic cells induces Th1 and Th17 cell differentiation. Mol. Immunol. 47:922–31.CrossRefPubMedGoogle Scholar
  37. 37.
    Chu P, Wierda WG, Kipps TJ. (2000) CD40 activation does not protect chronic lymphocytic leukemia B cells from apoptosis induced by cytotoxic T lymphocytes. Blood. 95:3853–8.PubMedGoogle Scholar
  38. 38.
    Sinistro A, et al. (2007) Lipopolysaccharide desensitizes monocytes-macrophages to CD40 ligand stimulation. Immunology 122:362–70.CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Banchereau J, Steinman RM. (1998) Dendritic cells and the control of immunity. Nature. 392:245–52.CrossRefGoogle Scholar
  40. 40.
    Osada T, Clay TM, Woo CY, Morse MA, Lyerly HK. (2006) Dendritic cell-based immunotherapy. Int. Rev. Immunol. 25:377–413.CrossRefPubMedGoogle Scholar
  41. 41.
    Banerjee DK, Dhodapkar MV, Matayeva E, Steinman RM, Dhodapkar KM. (2006) Expansion of FOXP3high regulatory T cells by human dendritic cells (DCs) in vitro and after injection of cytokine-matured DCs in myeloma patients. Blood. 108:2655–61.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© The Feinstein Institute for Medical Research 2011

Authors and Affiliations

  • Diahnn Futalan
    • 1
  • Chien-Tze Huang
    • 1
  • Ingo G. H. Schmidt-Wolf
    • 2
  • Marie Larsson
    • 3
  • Davorka Messmer
    • 1
  1. 1.Moores Cancer CenterUniversity of California San DiegoLa JollaUSA
  2. 2.Department of Internal MedicineRheinische Friedrich-Wilhelms UniversitaetBonnGermany
  3. 3.Division of Molecular Virology, Department of Clinical and Experimental MedicineLinköping UniversityLinköpingSweden

Personalised recommendations