Generation of peptide-specific cytotoxic T lymphocytes using allogeneic dendritic cells capable of lysing human pancreatic cancer cells.
BACKGROUND
Dendritic cells (DCs) are potent antigen presenting cells (APCs), able to efficiently induce primary T cell-mediated responses to foreign antigens. In earlier studies we were able to identify a histocompatibility antigen (HLA)-A 2-restricted nine amino acid peptide (GP2, peptide 654-662) from the transmembrane portion of the protooncogene HER2/neu as a tumor-associated antigen (TAA) in human pancreatic cancer.


METHODS
Peripheral blood mononuclear cells (PBMCs) of HLA-A2+ and HLA-A2 healthy volunteers were isolated. PBMCs were grown with initial anti-CD3, low-dose interleukin-2 (IL-2), and peptide-pulsed DC stimulation. T-cell lines were analyzed in functional studies.


RESULTS
After 4 weeks, T-cell cultures were more than 50% CD8+. All peptide-pulsed T cells significantly lysed APC pulsed with the immunizing antigen in an HLA-A2 restricted fashion. Furthermore, HLA-A2+,HER2/neu+ human pancreatic cancer cells were lysed significantly higher than HLA-A2 HER2/neu+ pancreatic cancer cells. Transfection of an HLA-A2 pancreatic cancer cell line with the HLA-A2 gene resulted in a significantly higher lysis of the transfected cell line compared to the wild type. In HLA-A2+ pancreatic cancer targets, specific lysis was HLA-A2 restricted.


CONCLUSIONS
The ability to use DCs for presentation of either tumor or peptide antigen in an HLA-restricted fashion to stimulate T-cell proliferation, as well as cytotoxicity, demonstrates the potential of this technology for future development of a pancreatic cancer vaccine.
Peiper
M
M
Department of Surgery, Brigham and Women's Hospital, Harvard Medical School, Boston, Mass. 02115, USA.
Goedegebuure
P S
PS
Eberlein
T J
TJ
D000595:Amino Acid Sequence; D017252:CD3 Complex / Q000276:immunology; D016516:CD4-CD8 Ratio; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology*; D015789:HLA-A2 Antigen / Q000097:blood / Q000276:immunology; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology; D010190:Pancreatic Neoplasms / Q000276:immunology* / Q000473:pathology*; D010446:Peptide Fragments / Q000494:pharmacology*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000737:chemistry / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects / Q000276:immunology*; D014407:Tumor Cells, Cultured
D017252:CD3 Complex; D015789:HLA-A2 Antigen; D007376:Interleukin-2; D010446:Peptide Fragments; D018719:Erb-b2 Receptor Tyrosine Kinases