In vitro analysis of T cell responses induced by breast tumor cell lysate pulsed with autologous dendritic cells

Abstract

In this In vitro study, T cell responses induced by breast tumor cell lysate pulsed monocyte-derived DCs were analyzed in terms of proliferation, specific cytotoxicity and cytokine-release in order to use in immunotherapeutic settings. Nylon wool enriched T lymphocytes from 5 patients with breast cancer stimulated In vitro with tumor cell lysate pulsed monocyte-derived DCs and their proliferation response were analyzed by [3H] thymidine uptake test. Specific cytotoxic activity of tumor antigen primed T cells after three rounds weekly stimulation was evaluated by flow cytometry, and interferon-γ (IFN-γ) and interleukin-4 (IL-4) cytokines release assay was carried out 24 hours after last stimulation in the supernatant of primed T cells using commercially available ELI-SA kits. T cell proliferation assay revealed that tumor cell lysate pulsed DCs could stimulate autologous T cell proliferation response with stimulation indices 4.9 - 30. T cell mediated cytotoxicity assay demonstrated that tumor antigen primed T cells could significantly kill autologous tumor cells more than normal cells (P < 0.05). These cells had variable amounts of cytotoxic activity against K562 cells as well. Primed T cells were released both IFN-γ and IL-4 in response to restimulation by antigen pulsed DCs which were dominated by IFN-γ production in 2 and IL-4 production in 3 out of 5 patients. Our result suggested that breast tumor antigen pulsed DCs could elicit effective specific antitumor T cell responses In vitro, therefore, tumor antigen pulsed DC vaccination may be considered as a novel strategy for immunotherapy of patients with breast cancer refractory to standard modalities.

Share and Cite:

Delirezh, N. , Moazzeni, S. , Shokri, F. , Shokrgozar, M. , Morteza Atri, M. and Karbassian, H. (2012) In vitro analysis of T cell responses induced by breast tumor cell lysate pulsed with autologous dendritic cells. Advances in Bioscience and Biotechnology, 3, 126-136. doi: 10.4236/abb.2012.32019.

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1] Ries, L.A.G., Kosary, C.L., Hankey, B.F. and Miller, B.A. (1998) SEER cancer statistics review, 1973-1995. National Cancer Institute, Bethesda.
[2] Berinstein, N. (2003) Overview of therapeutic vaccination approaches for cancer. Seminars in Oncology, 30, 1-8.
[3] Steinman, R.M. (1991) The dendritic cell system and its role in immunogenicity. Annual Review of Immunology, 9, 271-296. doi:10.1146/annurev.iy.09.040191.001415
[4] Cella, M., Sallusto, F. and Lanzavecchia, A. (1997) Origin, maturation and antigen presenting function of dendritic cells. Current Opinion in Immunology, 9, 10-16. doi:10.1016/S0952-7915(97)80153-7
[5] Brossart, P., Zobywalski, A., Grunebach, F., et al. (2000) Tumor necrosis factor alpha and CD40 ligand antagonize the inhibitory effects of interleukin-10 on T-cell stimulatory capacity of dendritic cells. Cancer Research, 60, 4485-4492.
[6] Labeur, M.S., Roters, B., Pers, B., Mehling, A., Luger, T.A., Schwan, T. and Grabbe, S. (1999) Generation of tumor immunity by bone marrow-derived dendritic cells correlated with dendritic cell maturation stage. Journal of Immunology, 162, 168-175.
[7] McDermott, R.S., Beuvon, F., Pauly, M., et al. (2002) Tumor antigen and antigen-presenting capacity in breast cancer. Pathobiology, 3, 324-332. doi:10.1159/000071272
[8] Tsuge, T., Yamakawa, M. and Tsukamoto, M. (2000) Infiltrating dendritic/Langerhans cells in primary breast cancer. Breast Cancer Research and Treatment, 59, 141-152. doi:10.1023/A:1006396216933
[9] Fadul, C.E., Fisher, J.L., Hampton, T.H., et al. (2011) Immune response in patients with newly diagnosed glioblastoma multiforme treated with intranodal autologous tumor lysate-dendritic cell vaccination after radiation chemotherapy. Journal of Immunotherapy, 34, 382- 389. doi:10.1097/CJI.0b013e318215e300
[10] Soleimani, A., Berntsen, A., Svane, I.M. and Pedersen, A.E. (2009) Immune responses in patients with metastatic renal cell carcinoma treated with dendritic cells pulsed with tumor lysate. Scandinavian Journal of Immunology, 70, 481-489. doi:10.1111/j.1365-3083.2009.02322.x
[11] Schwaab, T., Schwarzer, A., Wolf, B., et al. (2009) Clinical and immunologic effects of intranodal autologous tumor lysate-dendritic cell vaccine with Aldesleukin (Interleukin 2) and IFN-{alpha} 2a therapy in metastatic renal cell carcinoma patients. Clinical Cancer Research, 15, 4986-4992.
[12] Friedl, J., Stift, A., Poalini, P., et al. (2000) Tumor antigen pulsed dendritic cells enhance the cytolytic activity of tumor infilterating lymphocytes in human hepatocellular cancer. Cancer Biotherapy and Radiopharmaceuticals, 15, 477-486. doi:10.1089/cbr.2000.15.477
[13] Gilboa, E., Nair, S.K. and Lyerly, H.K. (1998) Immunotherapy of cancer with dendritic cell-based vaccine. Cancer Immunology, Immunotherapy, 46, 82-87. doi:10.1007/s002620050465
[14] Thurner, B., Roder, C., Dieckmann, D., et al. (1999) Generation of large numbers of fully mature and stable dendritic cells from leukaphersis product for clinical application. Journal of Immunological Methods, 223, 1-15. doi:10.1016/S0022-1759(98)00208-7
[15] Romani, N., Gruner S., Brang, D., et al. (1994) Proliferating dendritic cell progenitors in human blood. Journal of Experimental Medicine, 180, 83-93. doi:10.1084/jem.180.1.83
[16] Vilella, R., Benitez, D., Mila, J., et al. (2004) Pilot study of treatment of biochemotherapy—Refractory stage IV melanoma patients with autologous dendritic cells pulsed with a heterologous melanoma cell line lysate. Cancer Immunology, Immunotherapy, 53, 651-665. doi:10.1007/s00262-003-0495-3
[17] Paglia, P., Chiodoni, C., Rodolfo, M. and Colombo, M.P. (1996) Murine dendritic cells loaded in vitro with soluble protein prime cytotoxic T lymphocytes against tumor antigen in vivo. Journal of Experimental Medicine, 183, 317-322. doi:10.1084/jem.183.1.317
[18] Goldsmid, R.S., Idoyaga, J., Bravo, A.L., Steinman, R., Mordoh, J. and Wainstok, R. (2003) Dendritic cells charged with apoptotic tumor cells induce long-lived protective CD4+ and T cell immunity against B16 melanoma. Journal of Immunology, 171, 5940-5947.
[19] Zitvogel, L., Regnault, A., Lozier, A., et al. (1998) Eradicatian of established murine tumors using a novel cell free vaccine, dendritic cell derived exosomes. Nature, 4, 594-600. doi:10.1038/nm0598-594
[20] Galea-Lauri, J., Darling, D., Mufti, G., Harrison, P. and Farzaneh, F. (2002) Eliciting cytotoxic T lymphocytes against acute myeloid leukemia-derived antigen: Evaluation of dendritic cell-leukemia cell hybrids and other antigen-loading strategies for dendritic cell-based vaccination. Cancer Immunology, Immunotherapy, 51, 299-310. doi:10.1007/s00262-002-0284-4
[21] Boczkowski, D., Nair, S.K., Snyder, D. and Gilboa, E. (1996) Dendritic cell pulsed with RNA are potent antigen presenting cells in vitro and in vivo. Journal of Experimental Medicine, 184, 465-472. doi:10.1084/jem.184.2.465
[22] Inaba, K., Metlay, T.P., Crowely, M.T. and Steinman, R.M. (1990) Dendritic cells pulsed with protein antigen in vitro can prime antigen specific MHC-restricted T cells in situ. Journal of Experimental Medicine, 172, 631-640. doi:10.1084/jem.172.2.631
[23] Speirs, A.R., Waiton, D.S. and Atkin, S.L. (1998) Short- Term primary culture of epithelial cells derived from human breast tumors. British Journal of Cancer, 78, 1421-1429. doi:10.1038/bjc.1998.702
[24] Gong, J., Avigan, D., Chen, D., Wu, Z., Koido, S., Kashiwaba, M. and Kufe, D. (1999) Activation of antitumor cytotoxic T lymphocytes by fusion of human dendritic cells and breast carcinoma cells. Proceedings of the National Academy of Sciences USA, 97, 2715-2718. doi:10.1073/pnas.050587197
[25] Fischer, K. and Mackensen, A. (2003) The flow cytometric PKH-26 assay for the determination of T cell mediated cytotoxic activity. Methods, 31, 135-142. doi:10.1016/S1046-2023(03)00123-3
[26] Huang, A.Y., Golumbek, P., Ahmadzadeh, M., Jaffee, E., Paradoll, D. and Levitsky, H. (1994) Role of bone marrow-derived cells in presenting MHC class I-restricted tumor antigens. Science, 264, 961-965. doi:10.1126/science.7513904
[27] Banchereau, J., Schuler, B., Thurner, A., Palucka, K. and Schuler, G. (2001) Dendritic cells as vectors for therapy. Cell, 106, 271-274. doi:10.1016/S0092-8674(01)00448-2
[28] Wu, Y.G., Wu, G.Z., Wang, L., Zhang, Y.Y., Li, Z. and Li, D.C. (2010) Tumor cell lysate-pulsed dendritic cells induce a T cell response against colon cancer in vitro and in vivo. Medical Oncology, 27, 736-742. doi:10.1007/s12032-009-9277-x
[29] Peoples, G.E., Goedegeboure, P.S., Smith, R., Linehan, D.C., Yoshimo, I. and Eberlein, T.J. (1995) Breast and ovarian cancer specific cytotoxic T lymphocytes recognize the same HER 2/neu-derived peptide. Proceedings of the National Academy of Sciences USA, 92, 432-436. doi:10.1073/pnas.92.2.432
[30] Kass, R., Agha, J., Bellone, S., et al. (2003) In vitro induction of tumor specific HLA class I-restricted CD8+ cytotoxic T lymphocytes from patients with locally advanced breast cancer by antigen pulsed autologous dendritic cells. Journal of Surgical Research, 112, 189-197. doi:10.1016/S0022-4804(03)00147-1
[31] Kobayashi, T., Shinohara, H., Toyoda, M., Iwamoto, S. and Tanigawa, N. (2001) Regression of lymph node metastases by immunotherapy using autologous breast tumor lysate pulsed dendritic cells, report of a case. Surgery Today, 31, 513-516. doi:10.1007/s005950170112
[32] Kokhaei, P., Rezvany, M.R., Virving, L., Choudhury, A., Rabbani, H., Osterborg, A. and Mellstedt, H. (2003) Dendritic cells loaded with apoptotic tumor cells induce a stronger T cell response that dendritic cell tumor hybrids in B-CLL. Leukemia, 17, 894-899. doi:10.1038/sj.leu.2402913
[33] Gutza, E. and Okada, C.Y. (2002) Tumor cell lysate pulsed dendritic cells are more effective than Id protein vaccines for active immunotherapy of T cell lymphoma. Journal of Immunology, 169, 5227-5235.
[34] Schnurr, M., Scholz, C., Rothenfusser, Galambos, P., Duuer, M., Robe, J. and Endres, S. (2002) Apoptotic pancreatic tumor cells are superior to cell lysate in promoting cross priming of cytotoxic T cells and active NK and γ δ T cells. Cancer Research, 62, 2347-2352.
[35] Hayashi, T., Hideshima, T., Akiyama, M., Raje, N., Richardson, P., Chauhan, D. and Anderson, K.C. (2003) Ex vivo induction of multiple myeloma-specific cytotoxic T lymphocytes. Journal of Experimental Medicine, 102, 1435-1442.
[36] Fields, R.C., Shimizu, K. and Mule, J.J. (1998) Murine dendrtic cell pulsed with whole tumor lysates mediate potent antitumor immune responses in vitro and in vivo. Proceedings of the National Academy of Sciences USA, 95, 9482-9487. doi:10.1073/pnas.95.16.9482
[37] Asavaroengchai, W., Kotera, Y. and Mule, J.J. (2002) Tumor lysate pulsed dendritic cells can elicit an effective antitumor immune response during early lymphoid recovery. Proceedings of the National Academy of Sciences USA, 99, 931-936. doi:10.1073/pnas.022634999
[38] Ghosh, P., Komschlies, K.L., Cippitelli, M., Longo, D.L. and Sublesli, J. (1995) Gradual loss of T-helper 1 populations in spleen of mice during progressive tumor growth. Journal of the National Cancer Institute, 87, 1478-1483. doi:10.1093/jnci/87.19.1478
[39] Santin, A.D., Bellone, S., Ravaggi, A., Pecorelli, S., Cannon, M.J. and Parham, G.P. (2000) Induction of ovarian tumor specific CD8+ cytotoxic T lymphocytes by acideluted peptide-pulsed autologous dendritic cells. Obstetrics & Gynecology, 96, 422-430. doi:10.1016/S0029-7844(00)00916-9
[40] Rieser, C., Romaner, R., Holtl, L. and Rogtsch, H. (1999) Mature dendritic cells induce T helper type 1 dominant immune response in patients with renal cell carcinoma. Urologia Internationalis, 63, 151-159. doi:10.1159/000030438
[41] Bachleitner-Hofmann, T., Friedl, J., Hassler, M., et al. (2009) Pilot trial of autologous dendritic cells loaded with tumor lysate(s) from allogeneic tumor cell lines in patients with metastatic medullary thyroid carcinoma. Oncology Reports, 21, 1585-1592. doi:10.3892/or_00000391
[42] Kim, J.H., Lee, Y., Bae, Y.S., et al. (2007) Phase I/II study of immunotherapy using autologous tumor lysate-pulsed dendritic cells in patients with metastatic renal cell carcinoma. Clinical Immunology, 125, 257-267. doi:10.1016/j.clim.2007.07.014

Copyright © 2024 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.