부산대학교 도서관

Byline: Andreas Mackensen (1), Hendrik Veelken (1), Michael Lahn (1), Sebastian Wittnebel (1), Daniel Becker (1), Gabriele Kohler (2), Peter Kulmburg (1), Ulrich Brennscheidt (1), Felicia Rosenthal (1), Brigitte Franke (1), Roland Mertelsmann (1), A. Lindemann (1) Keywords: Key wordsaGene Therapy; Vaccination; Melanoma; Cytotoxic T lymphocytes; Tumor-infiltrating lymphocytes Abstract: aIn 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.sup.+ ]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.sup.+ ]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. Author Affiliation: (1) Department of Hematology/Oncology, University of Freiburg Medical Center, Hugstetterstrasse 55, D-79106 Freiburg, Germany, DE (2) Department of Pathology, University of Freiburg Medical Center, Hugstetterstrasse 55, D-79106 Freiburg, Germany, DE Article note: Received: 1 October 1996 / Accepted: 9 December 1996