Abstract
In order to understand further the effects of Newcastle-disease-virus(NDV)-modified tumour vaccines we investigated the feasibility of isolating lymphocytes from the site of injection of patients undergoing postoperative active specific immunization (ASI) with autologous NDV-modified tumour cells. Delayed-type-hypersensitivity(DTH)-like reactions from five cancer patients were surgically removed, minced and the tissue particles were digested with collagenase and DNase. Lymphoid cells recovered were expanded in a highly efficient limiting-dilution analysis system optimized for T cell growth [Moretta et al. (1983) J Exp Med 157: 743] and lymphocyte microcultures (clonal probability >0.8) could be grown for up to 1 year. Analysis of the microcultures for phenotype and function showed that the majority were positive for CD4 (92%) and TCRαβ (96%). Concanavalin-A-induced production of interleukin-2 (IL-2), IL-6, interferon γ and tumour necrosis factor α was detected in more than 70% of the microcultures. Lectin-dependent cytotoxicity was only very rarely observed. The general characteristics of the microcultures obtained support the notion of a DTH-like reaction taking place at the site of tumour cell challenge. The possibility of in vitro expansion and cultivation of T lymphocytes from ASI vaccination sites should help to elucidate further the role of these cells in active specific immunization against autologous tumour cells.
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Bier H, Armonat G, Bier J, Schirrmacher V, Ganzer U (1989) Postoperative active-specific immunotherapy of lymph node micrometastasis in a guinea pig tumor model. Otorhinolaryngology 51: 197
Bohle W, Schlag P, Liebrich W, Hohenberger P, Manasterski M, Müller P, Schirrmacher V (1990) Postoperative active specific immunization in colorectal cancer patients with virus-modified autologous tumor-cell vaccine: first clinical results with tumor cell vaccines modified with live but avirulent newcastle disease virus. Cancer 66: 1517
Dvorak HF, Galli SJ, Dvorak AM (1986) Cellular and vascular manifestations of cell-mediated immunity. Hum Pathol 17: 122
Fleischer B (1988) Non-specific propagation of human antigen-dependent T lymphocyte clones. J Immunol Methods 109: 215
Foon KA (1989) Biological response modifiers: the new immunotherapy. Cancer Res 49: 1621
Heicappell R, Schirrmacher V, von Hoegen P, Ahlert T, Appelhans B (1986) Prevention of metastatic spread by postoperative immunotherapy with virally modified autologous tumor cells. I. parameters for optimal therapeutic effect. Int J Cancer 37: 569
Ijzermans JN, Marquet RL (1989) Interferon-gamma: a review. Immunobiology 179: 456
Lehner B, Schlag P, Liebrich W, Schirrmacher V (1990) Postoperative active specific immunization in curatively resected colorectal cancer patients with a virus-modified autologous tumor cell vaccine. Cancer Immunol Immunother 32: 173
Liebrich W, Schlag P, Manasterski M, Lehner B, Stöhr M, Müller P, Schirrmacher V (1991) In vitro and clinical characterisation of a newcastle disease virus-modified autologous tumor cell vaccine for treatment of colorectal cancer patients. Eur J Cancer 27: 703
Männel DN, Falk W (1989) Optimal induction of tumor necrosis factor production in human monocytes requires complete S-form lipopolysaccharide. Infect Immun 57: 1953
Miescher S, Stoeck M, Qiao L, Barras C, Barrelet L, von Fliedner V (1988) Proliferative and cytolytic potentials of purified human tumor-infiltrating T-lymphocytes. Impaired response to mitogendriven stimulation despite T-cell receptor expression. Int J Cancer 42: 659
Moretta A, Pantaleo G, Moretta L, Cerottini JC, Mingari MC (1983) Direct demonstration of the clonogenic potential of every human peripheral blood T cell. Clonal analysis of HLA-DR expression and cytolytic activity. J Exp Med 157: 743
Rosenberg SA, Schwarz SL, Spiess PJ (1988) Combination immunotherapy for cancer: Synergistic antitumor interactions of interleukin-2, alpha interferon and tumor-infiltrating lymphocytes. JNCI 80: 1393
Schirrmacher V (1993) Immunity and metastasis: in situ activation of protective T cells by virus modified cancer vaccines. Cancer Surv (in press)
Schirrmacher V, Heicappel R (1987) Prevention of metastatic spread by postoperative immunotherapy with virally modified autologous tumor cells. II. Establishment of specific systemic anti-tumor immunity. Clin Exp Metastasis 5: 147
Schirrmacher V, Ahlert T, Heicappell R, Appelhans B, von Hoegen P (1986) Successful application of non-oncogenic viruses for antimetastatic cancer immunotherapy. Cancer Rev 5: 19
Schirrmacher V, von Hoegen P, Schlag P, Liebrich W, Lehner B, Schumacher K, Ahlert T, Bastert G (1989) Active specific immunotherapy with autologous tumor cell vaccines modified by Newcastle disease virus: experimental and clinical studies. In: Schirrmacher V, Schwartz-Albiez R (eds) Cancer metastasis. Springer, Berlin Heidelberg New York, pp 157–170
Shoham J, Hirsch R, Zakay-Rones Z, Osband M, Brennert HJ (1990) Augementation of tumor cell immunogenicity by viruses-an approach to specific immunotherapy of cancer. Nat Immun Cell Growth Regul 9: 165
Stevenson HC, Foon KA, Kanapa DJ, Favilla T, Beman I, Oldham RK (1984) The potential value of cytapheresis for adoptive immuntherapy of cancer patients. Plasma Ther Transf Technol 5: 237
Weber E, Kees UR, Krammer P Computer analysis of data from limiting dilution experiments of cells of the immune system. DKFZ Research Monograph
Whiteside TL, Miescher S, Hurlimann J, Moretta L, von Fliedner V (1986) Separation, phenotyping and limiting dilution analysis of T-lymphocytes infiltrating human solid tumors. Int J Cancer 37: 803
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This study was supported by Dr.-Mildred-Scheel-Stiftung and the Tumorzentrum Heidelberg
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Stoeck, M., Marland-Noske, C., Manasterski, M. et al. In vitro expansion and analysis of T lymphocyte microcultures obtained from the vaccination sites of cancer patients undergoing active specific immunization with autologous Newcastle-disease-virus-modified tumour cells. Cancer Immunol Immunother 37, 240–244 (1993). https://doi.org/10.1007/BF01518517
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DOI: https://doi.org/10.1007/BF01518517