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 |
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Authors: | M. Stoeck C. Marland-Noske M. Manasterski R. Zawatzky S. Horn V. Möbus P. Schlag V. Schirrmacher |
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Affiliation: | (1) German Cancer Research Centre, Institut für Immunologie und Genetics, Heidelberg, Germany;(2) Institute for Virus Research, German Cancer Research Centre, Heidelberg, Germany;(3) Section of Surgical Oncology, Department of Surgery, University of Heidelberg, Germany;(4) Department of Gynaecology and Obstetrics, University of Mainz, Germany;(5) Institut für Immunologie und Genetik/710, DKFZ, Im Neuenheimer Feld 280, D-6900 Heidelberg, Germany |
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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 TCR (96%). Concanavalin-A-induced production of interleukin-2 (IL-2), IL-6, interferon and tumour necrosis factor 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.This study was supported by Dr.-Mildred-Scheel-Stiftung and the Tumorzentrum Heidelberg |
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Keywords: | Active specific immunization NDV-modified tumour cells Microcultures Tumour vaccines |
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