Medical Oncology

, Volume 27, Issue 3, pp 736–742 | Cite as

Tumor cell lysate-pulsed dendritic cells induce a T cell response against colon cancer in vitro and in vivo

Original Paper

Abstract

To investigate whether tumor cell lysate-pulsed (TP) dendritic cells (DCs) induce cytotoxic T lymphocyte (CTL) activity against colon cancer in vitro and in vivo. Hematopoietic progenitor cells were magnetically isolated from BALB/c mice bone marrow cells. These cells were cultured with cytokines GM-CSF, IL-4, and TNFα to induce their maturation. They were analyzed by morphological observation and phenotype analysis. DCs were pulsed with tumor cell lysate obtained by rapid freezing and thawing at a 1:3 DC:tumor cell ratio. CTL activity and interferon gamma (IFNγ) secretion was evaluated ex vivo. In order to determine whether or not vaccination with CT26 TP DCs induce the therapeutic potential in the established colon tumor model, CT26 colon tumor cells were implanted subcutaneously (s.c.) in the midflank of naïve BALB/c mice. Tumor-bearing mice were injected with vaccination with CT26 TP DCs on days 3 and 10. Tumor growth was assessed every 2–3 days. Finally, CTL activity and IFNγ secretion were evaluated in immunized mice. Hematopoietic progenitor cells from mice bone marrow cells cultured with cytokines for 8 days showed the character of typical mature DCs. Morphologically, these cells were large with oval or irregularly shaped nuclei and with many small dendrites. Phenotypically, FACS analysis showed that they expressed high levels of MHC II, CD11b, CD80, and CD86 antigen, and were negative for CD8α. However, immature DCs cultured with cytokines for 5 days did not have typical DCs phenotypic markers. Ex vivo primed T cells with CT26 TP DCs were able to induce effective CTL activity against CT26 tumor cells, but not B16 tumor cells (E:T = 100:1, 60.36 ± 7.11% specific lysis in CT26 group vs. 17.36 ± 4.10% specific lysis in B16 group), and produced higher levels of IFNγ when stimulated with CT26 tumor cells but not when stimulated with B16 tumor cells (1210.33 ± 72.15 pg/ml in CT26 group vs. 182.25 ± 25.51 pg/ml in B16 group, P < 0.01). Vaccination with CT26 TP DCs could induce anti-tumor immunity against CT26 colon tumor in murine therapeutic models (tumor volume on day 19: CT26 TP DCs 342 ± 55 mm3 vs. the other control groups, P < 0.05). In addition, all splenic CD3+ T cells obtained from mice vaccinated with CT26 TP DCs produced high levels of IFNγ and shown specific cytotoxic activity against CT26 tumor cells, but no cytotoxic activity when stimulated with B16 tumor cells. Tumor cell lysate-pulsed DCs can induce tumor-specific CTL activity against colon cancer in vitro and in vivo.

Keywords

Vaccination Dendritic cells Cytotoxic T lymphocyte Colon cancer Immunotherapy 

References

  1. 1.
    Rougier P, Andre T, Panis Y, Colin P, Stremsdoerfer N, Laurent-Puig P. Colon cancer. Gastroenterol Clin Biol. 2006;30(2):2S24–9. PMID: 17151560.PubMedGoogle Scholar
  2. 2.
    Mano MS, Duhoux F. Colon cancer: update on adjuvant therapy. Clin Colorectal Cancer. 2008;7(3):178–83. doi: 10.3816/CCC.2008.n.023. PMID: 18621635.CrossRefPubMedGoogle Scholar
  3. 3.
    Xu R, Zhou B, Fung PC, Li X. Recent advances in the treatment of colon cancer. Histol Histopathol. 2006;21(8):867–72. PMID: 16691539.PubMedGoogle Scholar
  4. 4.
    Morse M, Langer L, Starodub A, Hobeika A, Clay T, Lyerly HK. Current immunotherapeutic strategies in colon cancer. Surg Oncol Clin N Am. 2007;16(4):873–900. doi: 10.1016/j.soc.2007.07.005. PMID: 18022550.CrossRefPubMedGoogle Scholar
  5. 5.
    Giboa E. DC-based cancer vaccines. J Clin Invest. 2007;117:1195–203. doi: 10.1172/JCI31205. PMID: 17476349.CrossRefGoogle Scholar
  6. 6.
    Wu Y, Wang L, Zhang Y. Dendritic cells as vectors for immunotherapy of tumor and its application for gastric cancer therapy. Cell Mol Immunol. 2004;1:351–6. PMID:16285894.PubMedGoogle Scholar
  7. 7.
    Aarntzen EH, Figdor CG, Adema GJ, Punt CJ, de Vries IJ. Dendritic cell vaccination and immune monitoring. Cancer Immunol Immunother. 2008;57(10):1559–68. doi: 10.1007/s00262-008-0553-y. PMID: 18618110.CrossRefPubMedGoogle Scholar
  8. 8.
    Nouri-Shirazi M, Banchereau J, Fay J, Palucka K. Dendritic cell based tumor vaccines. Immunol Lett. 2000;74(1):5–10. doi: 10.1016/S0165-2478(00)00243-1. PMID: 10996622.CrossRefPubMedGoogle Scholar
  9. 9.
    Li YL, Wu YG, Wang YQ, Li Z, Wang RC, Wang L, et al. Bone marrow-derived dendritic cells pulsed with tumor lysates induce anti-tumor immunity against gastric cancer ex vivo. World J Gastroenterol. 2008;14(46):7127–32. doi: 10.3748/wjg.14.7127. PMID: 19084922.CrossRefPubMedGoogle Scholar
  10. 10.
    Bachleitner-Hofmann T, Strohschneider M, Krieger P, Sachet M, Dubsky P, Hayden H, et al. Heat shock treatment of tumor lysate-pulsed dendritic cells enhances their capacity to elicit antitumor T cell responses against medullary thyroid carcinoma. J Clin Endocrinol Metab. 2006;91(11):4571–7. doi: 10.1210/jc.2006-0971. PMID: 16954161.CrossRefPubMedGoogle Scholar
  11. 11.
    Zhang Y, Harada A, Wang JB, Zhang YY, Hashimoto S, Naito M, et al. Bifurcated dendritic cell differentiation in vitro from murine lineage phenoltype-negative c-kit+ bone marrow hematopoitetic progenitor cells. Blood. 1998;92:118–28. PMID: 9639507.PubMedGoogle Scholar
  12. 12.
    Zhang Y, Zhang YY, Ogata M, Chen P, Harada A, Hashimoto S, et al. Transforming growth factor-betal polarizes murine hematopoietic progenitor cells to generate langerhans cell-like dendritic cells through a monocyte/macrophage differentiation pathway. Blood. 1999;93:1208–20. PMID: 9949163.PubMedGoogle Scholar
  13. 13.
    Lin KW, Jacek T, Jacek R. Dendritic cells heterogeneity and its role in cancer immunity. J Cancer Res Ther. 2006;2(2):35–40. PMID: 17998672.CrossRefPubMedGoogle Scholar
  14. 14.
    Ardavín C. Origin precursors and differentiation of mouse dendritic cells. Nat Rev Immunol. 2003;3(7):582–90. doi: 10.1038/nri1127. PMID: 12876560.CrossRefPubMedGoogle Scholar
  15. 15.
    Jacobs B, Wuttke M, Papewalis C, Seissler J, Schott M. Dendritic cell subtypes and in vitro generation of dendritic cells. Horm Metab Res. 2008;40(2):99–107. doi: 10.1055/s-2007-1022561. PMID: 18283627.CrossRefPubMedGoogle Scholar
  16. 16.
    Zou GM, Tam YK. Cytokines in the generation and maturation of dendritic cells: recent advances. Eur Cytokine Netw. 2002;13(2):186–99. PMID: 12101074.PubMedGoogle Scholar
  17. 17.
    Kondo H, Hazama S, Kawaoka T, Yoshino S, Yoshida S, Tokuno K, et al. Adoptive immunotherapy for pancreatic cancer using MUC1 peptide-pulsed dendritic cells and activated T lymphocytes. Anticancer Res. 2008;28(1B):379–87. PMID: 18383873.PubMedGoogle Scholar
  18. 18.
    Sas S, Chan T, Sami A, El-Gayed A, El-Gayed A, Xiang J. Vaccination of fiber-modified adenovirus-transfected dendritic cells to express HER-2/neu stimulates efficient HER-2/neu-specific humoral and CTL responses and reduces breast carcinogenesis in transgenic mice. Cancer Gene Ther. 2008;15(10):655–66. doi: 10.1038/cgt.2008.18. PMID: 18421311.CrossRefPubMedGoogle Scholar
  19. 19.
    Fukui M, Nakano-Hashimoto T, Okano K, Maruta Y, Suehiro Y, Hamanaka Y, et al. Therapeutic effect of dendritic cells loaded with a fusion mRNA encoding tyrosinase-related protein 2 and enhanced green fluorescence protein on B16 melanoma. Tumour Biol. 2004;25(5–6):252–7. doi: 10.1159/000081388. PMID: 15627888.CrossRefPubMedGoogle Scholar
  20. 20.
    Tamir A, Basagila E, Kagahzian A, Jiao L, Jensen S, Nicholls J, et al. Induction of tumor-specific T-cell responses by vaccination with tumor lysate-loaded dendritic cells in colorectal cancer patients with carcinoembryonic-antigen positive tumors. Cancer Immunol Immunother. 2007;56(12):2003–16. doi: 10.1007/s00262-007-0299-y. PMID: 17333181.CrossRefPubMedGoogle Scholar
  21. 21.
    Rosenberg SA, Marincola FM. Dendritic cells infected with poxviruses encoding MART-1/Melan A sensitize T lymphocytes in vitro. J Immunother. 1997;20(4):276–86. doi: 10.1097/00002371-199707000-00004. PMID: 9220317.CrossRefPubMedGoogle Scholar
  22. 22.
    Nair SK. Immunotherapy of cancer with dendritic cell-based vaccines. Gene Ther. 1998;5(11):1445–56. doi: 10.1038/sj.gt.3300790. PMID: 9930296.CrossRefPubMedGoogle Scholar
  23. 23.
    Gurnani K, Kennedy J, Sad S, Sprott GD, Krishnan L. Phosphatidylserine receptor-mediated recognition of archaeosome adjuvant promotes endocytosis and MHC class I cross-presentation of the entrapped antigen by phagosome-to-cytosol transport and classical processing. J Immunol. 2004;173(1):566–78. PMID: 15210818.PubMedGoogle Scholar
  24. 24.
    Palmer DH, Midgley RS, Mirza N, Torr EE, Ahmed F, Steele JC, et al. A phase II study of adoptive immunotherapy using dendritic cells pulsed with tumor lysate in patients with hepatocellular carcinoma. Hepatology. 2009;49(1):124–32. doi: 10.1002/hep.22626. PMID: 18980227.CrossRefPubMedGoogle Scholar
  25. 25.
    Chang AE, Redman BG, Whitfield JR, Nickoloff BJ, Braun TM, Lee PP, et al. A phase I trial of tumor lysate-pulsed dendritic cells in the treatment of advanced cancer. Clin Cancer Res. 2002;8(4):1021–32. PMID: 11948109.PubMedGoogle Scholar

Copyright information

© Humana Press Inc. 2009

Authors and Affiliations

  1. 1.Department of SurgeryThe First People Hospital of Changzhou and the Third Affiliated Hospital of Soochow UniversityChangzhouChina
  2. 2.Department of SurgeryThe First People Hospital of YanchengYanchengChina
  3. 3.Department of SurgeryThe First Affiliated Hospital of Soochow UniversitySuzhouChina
  4. 4.Institute of Health Sciences and Shanghai Institute of ImmunologyShanghai Institutes for Biological Sciences, Chinese Academy of Sciences and Shanghai Jiao Tong University School of MedicineShanghaiChina

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