Cancer Immunology, Immunotherapy

, Volume 55, Issue 6, pp 708–716 | Cite as

Early-outgrowth of endothelial progenitor cells can function as antigen-presenting cells

  • Masahiro AsakageEmail author
  • Nelson H. Tsuno
  • Joji Kitayama
  • Kazushige Kawai
  • Yurai Okaji
  • Kentaro Yazawa
  • Shoichi Kaisaki
  • Takuya Osada
  • Toshiaki Watanabe
  • Koki Takahashi
  • Hirokazu Nagawa
Original Article


Endothelial progenitor cells (EPCs) have been recently found to exist circulating in peripheral blood of adults, and home to sites of neovascularization in peripheral tissues. They can also be differentiated from peripheral blood mononuclear cells (PBMNCs). In tumor tissues, EPCs are found in highly vascularized lesions. Few reports exist in the literature concerning the characteristics of EPCs, especially related to their surface antigen expressions, except for endothelial markers. Here, we aimed to investigate the surface expression of differentiation markers, and the functional activities of early-outgrowth of EPCs (EO-EPCs), especially focusing on their antigen-presenting ability. EO-EPCs were generated from PBMNCs, by culture in the presence of angiogenic factors. These EO-EPCs had the morphological and functional features of endothelial cells and, additionally, they shared antigen-presenting ability. They induced the proliferation of allogeneic lymphocytes in a mixed-lymphocyte reaction, and could generate cytotoxic lymphocytes, with the ability to lyze tumor cells in an antigen-specific manner. The antigen-presenting ability of EO-EPCs, however, was weaker than that of monocyte-derived dendritic cells, but stronger than peripheral blood monocytes. Since EO-EPCs play an important role in the development of tumor angiogenesis, targeting EPCs would be an effective anti-angiogenic strategy. Alternatively, due to their antigen-presenting ability, EO-EPCs can be used as the effectors of anti-tumor immunotherapy. Since they share endothelial antigens, the activation of a cellular immunity against angiogenic vessels can be expected. In conclusion, EO-EPCs should be an interesting alternative for the development of new therapeutic strategies to combat cancer, either as the effectors or as the targets of cancer immunotherapy.


Early-outgrowth of endothelial progenitor cells Endothelial progenitor cells Angiogenesis Antigen presenting cells 



Endothelial progenitor cells


Early-outgrowth of endothelial progenitor cells


Endothelial cells


Peripheral blood mononuclear cells


Acidic fibroblast growth factor


1, 1′-Dioctadecyl-3, 3, 3′, 3′-tetramethylindocarbocyanine-labeled acetylated low-density lipoprotein


Antigen-presenting cells


Cytotoxic T lymphocytes


Monocyte-derived dendritic cells


Human umbilical vein endothelial cells


Fetal calf serum


Mixed lymphocytes reaction


Fluorescein isothiocyanate



This study was supported partly by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan, partly by a grant from the Ministry of Health, Labour and Welfare of Japan, and partly by a grant from the Sankyo Foundation of Life Science.


  1. 1.
    Albert ML, Pearce SF, Francisco LM, Sauter B, Roy P, Silverstein RL, Bhardwaj N (1998) Immature dendritic cells phagocytose apoptotic cells via alphavbeta5 and CD36, and cross-present antigens to cytotoxic T lymphocytes. J Exp Med 188:1359–1368PubMedCrossRefGoogle Scholar
  2. 2.
    Asahara T, Masuda H, Takahashi T, Kalka C, Pastore C, Silver M, Kearne M, Magner M, Isner JM (1999) Bone marrow origin of endothelial progenitor cells responsible for postnatal vasculogenesis in physiological and pathological neovascularization. Circ Res 85:221–228PubMedGoogle Scholar
  3. 3.
    Asahara T, Murohara T, Sullivan A, Silver M, van der Zee R, Li T, Witzenbichler B, Schatteman G, Isner JM (1997) Isolation of putative progenitor endothelial cells for angiogenesis. Science 275:964–967PubMedCrossRefGoogle Scholar
  4. 4.
    Asahara T, Takahashi T, Masuda H, Kalka C, Chen D, Iwaguro H, Inai Y, Silver M, Isner JM (1999) VEGF contributes to postnatal neovascularization by mobilizing bone marrow-derived endothelial progenitor cells. Embo J 18:3964–3972PubMedCrossRefGoogle Scholar
  5. 5.
    Asakage M, Kitayama J, Tsuno NH, Komuro Y, Kaisaki S, Hori N, Nagawa H, Takahashi K (2005) Primary malignant melanoma of the esophagus treated by esophagectomy and adjuvant dendritic-cell therapy. J Gastroenterol 40:545–546PubMedCrossRefGoogle Scholar
  6. 6.
    Asakage M, Tsuno NH, Kitayama J, Kawai K, Okaji Y, Yazawa K, Kaisaki S, Takahashi K, Nagawa H (2004) 3-Hydroxy-3-methylglutaryl-coenzyme A reductase inhibitor (pravastatin) inhibits endothelial cell proliferation dependent on G1 cell cycle arrest. Anticancer Drugs 15:625–632PubMedCrossRefGoogle Scholar
  7. 7.
    Banchereau J, Steinman RM (1998) Dendritic cells and the control of immunity. Nature 392:245–252PubMedCrossRefGoogle Scholar
  8. 8.
    Eggermann J, Kliche S, Jarmy G, Hoffmann K, Mayr-Beyrle U, Debatin KM, Waltenberger J, Beltinger C (2003) Endothelial progenitor cell culture and differentiation in vitro: a methodological comparison using human umbilical cord blood. Cardiovasc Res 58:478–486PubMedCrossRefGoogle Scholar
  9. 9.
    Fernandez Pujol B, Lucibello FC, Gehling UM, Lindemann K, Weidner N, Zuzarte ML, Adamkiewicz J, Elsasser HP, Muller R, Havemann K (2000) Endothelial-like cells derived from human CD14 positive monocytes. Differentiation 65:287–300PubMedCrossRefGoogle Scholar
  10. 10.
    Fernandez Pujol B, Lucibello FC, Zuzarte M, Lutjens P, Muller R, Havemann K (2001) Dendritic cells derived from peripheral monocytes express endothelial markers and in the presence of angiogenic growth factors differentiate into endothelial-like cells. Eur J Cell Biol 80:99–110PubMedCrossRefGoogle Scholar
  11. 11.
    Folkman J (1971) Tumor angiogenesis: therapeutic implications. N Engl J Med 285:1182–1186PubMedCrossRefGoogle Scholar
  12. 12.
    Folkman J, Shing Y (1992) Angiogenesis. J Biol Chem 267:10931–10934PubMedGoogle Scholar
  13. 13.
    Hancock WW, Sayegh MH, Zheng XG, Peach R, Linsley PS, Turka LA (1996) Costimulatory function and expression of CD40 ligand, CD80, and CD86 in vascularized murine cardiac allograft rejection. Proc Natl Acad Sci USA 93:13967–13972PubMedCrossRefGoogle Scholar
  14. 14.
    Hart PH, Whitty GA, Burgess DR, Croatto M, Hamilton JA (1990) Augmentation of glucocorticoid action on human monocytes by interleukin-4. Lymphokine Res 9:147–153PubMedGoogle Scholar
  15. 15.
    Holtl L, Rieser C, Papesh C, Ramoner R, Bartsch G, Thurnher M (1998) CD83+ blood dendritic cells as a vaccine for immunotherapy of metastatic renal-cell cancer. Lancet 352:1358PubMedCrossRefGoogle Scholar
  16. 16.
    Hsu FJ, Benike C, Fagnoni F, Liles TM, Czerwinski D, Taidi B, Engleman EG, Levy R (1996) Vaccination of patients with B-cell lymphoma using autologous antigen-pulsed dendritic cells. Nat Med 2:52–58PubMedCrossRefGoogle Scholar
  17. 17.
    Hur J, Yoon CH, Kim HS, Choi JH, Kang HJ, Hwang KK, Oh BH, Lee MM, Park YB (2004) Characterization of two types of endothelial progenitor cells and their different contributions to neovasculogenesis. Arterioscler Thromb Vasc Biol 24:288–293PubMedCrossRefGoogle Scholar
  18. 18.
    Inaba K, Inaba M, Naito M, Steinman RM (1993) Dendritic cell progenitors phagocytose particulates, including bacillus Calmette-Guerin organisms, and sensitize mice to mycobacterial antigens in vivo. J Exp Med 178:479–488PubMedCrossRefGoogle Scholar
  19. 19.
    Kalka C, Masuda H, Takahashi T, Kalka-Moll WM, Silver M, Kearney M, Li T, Isner JM, Asahara T (2000) Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization. Proc Natl Acad Sci USA 97:3422–3427PubMedCrossRefGoogle Scholar
  20. 20.
    Levin D, Constant S, Pasqualini T, Flavell R, Bottomly K (1993) Role of dendritic cells in the priming of CD4+ T lymphocytes to peptide antigen in vivo. J Immunol 151:6742–6750PubMedGoogle Scholar
  21. 21.
    Nestle FO, Alijagic S, Gilliet M, Sun Y, Grabbe S, Dummer R, Burg G, Schadendorf D (1998) Vaccination of melanoma patients with peptide- or tumor lysate-pulsed dendritic cells. Nat Med 4:328–332PubMedCrossRefGoogle Scholar
  22. 22.
    Page C, Rose M, Yacoub M, Pigott R (1992) Antigenic heterogeneity of vascular endothelium. Am J Pathol 141:673–683PubMedGoogle Scholar
  23. 23.
    Rehman J, Li J, Orschell CM, March KL (2003) Peripheral blood “endothelial progenitor cells” are derived from monocyte/macrophages and secrete angiogenic growth factors. Circulation 107:1164–1169PubMedCrossRefGoogle Scholar
  24. 24.
    Risau W, Sariola H, Zerwes HG, Sasse J, Ekblom P, Kemler R, Doetschman T (1988) Vasculogenesis and angiogenesis in embryonic-stem-cell-derived embryoid bodies. Development 102:471–478PubMedGoogle Scholar
  25. 25.
    Salgaller ML, Lodge PA, McLean JG, Tjoa BA, Loftus DJ, Ragde H, Kenny GM, Rogers M, Boynton AL, Murphy GP (1998) Report of immune monitoring of prostate cancer patients undergoing T-cell therapy using dendritic cells pulsed with HLA-A2-specific peptides from prostate-specific membrane antigen (PSMA). Prostate 35:144–151PubMedCrossRefGoogle Scholar
  26. 26.
    Seino K, Azuma M, Bashuda H, Fukao K, Yagita H, Okumura K (1995) CD86 (B70/B7-2) on endothelial cells co-stimulates allogeneic CD4+ T cells. Int Immunol 7:1331–1337PubMedCrossRefGoogle Scholar
  27. 27.
    Steinman RM (1991) The dendritic cell system and its role in immunogenicity. Annu Rev Immunol 9:271–296PubMedCrossRefGoogle Scholar
  28. 28.
    Urbich C, Heeschen C, Aicher A, Dernbach E, Zeiher AM, Dimmeler S (2003) Relevance of monocytic features for neovascularization capacity of circulating endothelial progenitor cells. Circulation 108:2511–2516PubMedCrossRefGoogle Scholar
  29. 29.
    Vallee I, Guillaumin JM, Thibault G, Gruel Y, Lebranchu Y, Bardos P, Watier H (1998) Human T lymphocyte proliferative response to resting porcine endothelial cells results from an HLA-restricted, IL-10-sensitive, indirect presentation pathway but also depends on endothelial-specific costimulatory factors. J Immunol 161:1652–1658PubMedGoogle Scholar
  30. 30.
    Vasa M, Fichtlscherer S, Adler K, Aicher A, Martin H, Zeiher AM, Dimmeler S (2001) Increase in circulating endothelial progenitor cells by statin therapy in patients with stable coronary artery disease. Circulation 103:2885–2890PubMedCrossRefGoogle Scholar
  31. 31.
    Zetter BR (1998) Angiogenesis and tumor metastasis. Annu Rev Med 49:407–424PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Masahiro Asakage
    • 1
    Email author
  • Nelson H. Tsuno
    • 1
    • 2
  • Joji Kitayama
    • 1
  • Kazushige Kawai
    • 1
  • Yurai Okaji
    • 1
  • Kentaro Yazawa
    • 1
  • Shoichi Kaisaki
    • 1
  • Takuya Osada
    • 1
  • Toshiaki Watanabe
    • 1
  • Koki Takahashi
    • 2
  • Hirokazu Nagawa
    • 1
  1. 1.Department of Surgical OncologyThe University of TokyoTokyoJapan
  2. 2.Department of Transfusion Medicine, Faculty of MedicineThe University of TokyoTokyoJapan

Personalised recommendations