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Lysis of human pancreatic adenocarcinoma cells by autologous hla‐class I‐restricted cytolytic T‐lymphocyte (CTL) clones
Author(s) -
Wölfel Thomas,
Herr Wolfgang,
Couue Pierre,
Schmitt Ute,
Zum Büschenfelde KarlHermann Meyer,
Knuth Alexander
Publication year - 1993
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.2910540419
Subject(s) - ctl* , cd8 , immunology , biology , t lymphocyte , cytolysis , antigen , lymphocyte , cytotoxic t cell , cytotoxicity , cd3 , tumor infiltrating lymphocytes , microbiology and biotechnology , in vitro , biochemistry
From the primary site of a pancreatic adenocarcinoma (patient BE) a permanent cell line (MZ‐PC‐2) was established in tissue culture. In the course of mixed lymphocyte‐tumor‐cell cultures (MLTC) with autologous blood‐derived lymphocytes, we isolated CTL clones that lysed autologous tumor cells but not autologous EBV‐transformed B cells (EBV‐B) and not K562. Pre‐treatment of MZ‐PC‐2 cells with IFN‐γ was required to obtain significant lysis in 4‐hr cytotoxicity assays. IFN‐γ was superior to IFN‐α in that respect. Among MLTC responder lymphocytes, tumor‐reactive CTL proliferated more strongly in response to MZ‐PC‐2 cells treated with IFN‐γ than to untreated tumor cells. Three CTL clones derived from MLTC were chosen for further analysis. They were CD3+, CD8+, TCR‐α/β + and behaved identically in all functional aspects tested. They all expressed the same TCR‐β chain, indicating that they descended from a common precursor lymphocyte and were directed against the same antigen. According to antibody‐inhibition experiments, BE‐CTL recognized their targets via an HLA‐B molecule carrying the Bw6 supertypic determinant. Irrespective of pre‐incubation with IFN‐γ, low levels of tumor‐cell lysis, or none, were seen when MZ‐PC‐2 cells were kept in medium supplemented with autologous serum or serum pooled from healthy volunteers instead of FCS. Lysability was restored when TNF‐α was added to human serum. Serum‐free medium was found to enhance the susceptibility of MZ‐PC‐2 cells to lysis by autologous CTL.