Generation of human tumor-reactive cytotoxic T cells against peptides presented by non-self HLA class I molecules.
The cyclin-D1 protein, which was found to be overexpressed in various human tumors, promotes cell cycle progression from the G1 into the S phase. It is normally expressed at low levels in several tissues and is likely to induce immunological tolerance. We have recently shown in a murine system that T cell tolerance to a widely expressed protein was circumvented by raising cytotoxic T lymphocytes (CTL) from MHC-mismatched donors. In this study, we tested whether it is possible to raise human allo-restricted CTL against the cyclin-D1 protein. The human cell line T2 is deficient in the genes encoding the transporter associated with antigen processing (TAP), resulting in inefficient loading of HLA-A2 class I molecules with endogenous peptides. Thus, a large number of A2 molecules can bind exogenously supplied synthetic peptides. Peripheral blood mononuclear cells from HLA-A2-negative donors were stimulated with T2 cells presenting cyclin-D1-derived synthetic peptides. Cloning of bulk cultures revealed that a large proportion of CTL clones were peptide specific. One peptide induced CTL which lysed cyclin-D1-expressing breast cancer cells, but not control Epstein-Barr virus-transformed B lymphoid cells. The results show that HLA-A2-negative donors can be used to isolate tumor-reactive CTL specific for cyclin-D1 peptides presented by HLA-A2 class I molecules.
Sadovnikova
E
E
Imperial College School of Medicine, Department of Immunology, Hammersmith Hospital, London, GB.
Jopling
L A
LA
Soo
K S
KS
Stauss
H J
HJ
D017951:Antigen Presentation; D001943:Breast Neoplasms / Q000473:pathology; D002461:Cell Line, Transformed; D019938:Cyclin D1 / Q000276:immunology*; D005260:Female; D015789:HLA-A2 Antigen / Q000276:immunology*; D006801:Humans; D006825:Hybridomas; D007519:Isoantigens / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology*; D011485:Protein Binding; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014407:Tumor Cells, Cultured
D015789:HLA-A2 Antigen; D007519:Isoantigens; D010446:Peptide Fragments; D019938:Cyclin D1