title: 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.
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 alpha beta (96%). Concanavalin-A-induced production of interleukin-2 (IL-2), IL-6, interferon gamma and tumour necrosis factor alpha 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.
authors.affiliation: Institute for Immunology and Genetics, German Cancer Research Centre, Heidelberg.
authors.forename: M
authors.initials: M
authors.lastname: Stoeck
authors.forename: C
authors.initials: C
authors.lastname: Marland-Noske
authors.forename: M
authors.initials: M
authors.lastname: Manasterski
authors.forename: R
authors.initials: R
authors.lastname: Zawatzky
authors.forename: S
authors.initials: S
authors.lastname: Horn
authors.forename: V
authors.initials: V
authors.lastname: Möbus
authors.forename: P
authors.initials: P
authors.lastname: Schlag
authors.forename: V
authors.initials: V
authors.lastname: Schirrmacher
mesh_terms: D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis; D003602:Cytotoxicity, Immunologic; D006801:Humans; D006968:Hypersensitivity, Delayed / Q000276:immunology; D016130:Immunophenotyping; D009369:Neoplasms / Q000276:immunology*; D009522:Newcastle disease virus; D013601:T-Lymphocytes / Q000276:immunology*; D014611:Vaccination / Q000379:methods*
chemical_list: D016207:Cytokines