title: Induction of tumor-specific cytotoxic T lymphocytes by immunization with autologous tumor cells and interleukin-2 gene transfected fibroblasts.
abstract: In a phase I trial designed to study a vaccine composed of autologous tumor cells and interleukin-2 gene transfected fibroblasts we analyzed lymphocytes infiltrating the vaccination site (VIL) in two melanoma patients. Functional studies demonstrated that numbers of MHC class I restricted cytotoxic T cells directed against the autologous tumor had increased at the immunization site in both cases. Analysis of the variability of T cell receptors (TCR) in the VIL of one patient revealed that the cytotoxic T lymphocytes consisted of a predominant population of TCRBV21S3+ T cells. Enrichment of this subpopulation to more than 99% by specific anti-TCRBV21S3 monoclonal antibody linked immunomagnetic beads and sequencing of the TCR-beta chain disclosed exactly the same V-D-J junctional sequence in all eight TCRBV21 transcripts from these VIL. The identical sequence was also detected in all eight TCRBV21 transcripts from the patient's tumor-infiltrating lymphocytes, indicating that the same CTL clone had infiltrated the tumor, circulated in the peripheral blood, and was amplified at the vaccination site. The TCRBV21S3+ T cells were also found to display an MHC class I restricted cytotoxic activity specifically directed against the autologous tumor cells. At the beginning of treatment these cells were undetectable at the vaccination site and delayed-type hypersensitivity testing was negative, contrasting with the positive results after therapy. Thus it is likely that vaccination with autologous tumor cells plus interleukin-2 gene transfected allogeneic fibroblasts had induced not only local accumulation but also an increase in the frequency of circulating tumor specific CTL.
authors.affiliation: Department of Hematology/Oncology, University of Freiburg Medical Center, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Mackensen
authors.forename: H
authors.initials: H
authors.lastname: Veelken
authors.forename: M
authors.initials: M
authors.lastname: Lahn
authors.forename: S
authors.initials: S
authors.lastname: Wittnebel
authors.forename: D
authors.initials: D
authors.lastname: Becker
authors.forename: G
authors.initials: G
authors.lastname: Köhler
authors.forename: P
authors.initials: P
authors.lastname: Kulmburg
authors.forename: U
authors.initials: U
authors.lastname: Brennscheidt
authors.forename: F
authors.initials: F
authors.lastname: Rosenthal
authors.forename: B
authors.initials: B
authors.lastname: Franke
authors.forename: R
authors.initials: R
authors.lastname: Mertelsmann
authors.forename: A
authors.initials: A
authors.lastname: Lindemann
mesh_terms: D000595:Amino Acid Sequence; D019496:Cancer Vaccines / Q000276:immunology / Q000627:therapeutic use*; D002460:Cell Line; D003602:Cytotoxicity, Immunologic; D005260:Female; D005347:Fibroblasts / Q000637:transplantation; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods; D007376:Interleukin-2 / Q000235:genetics*; D016246:Lymphocytes, Tumor-Infiltrating; D008545:Melanoma / Q000628:therapy*; D008875:Middle Aged; D009368:Neoplasm Transplantation; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000097:blood / Q000235:genetics; D012867:Skin / Q000276:immunology; D013601:T-Lymphocytes; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014611:Vaccination
chemical_list: D019496:Cancer Vaccines; D007376:Interleukin-2; D016693:Receptors, Antigen, T-Cell, alpha-beta