A TGFβRII frameshift-mutation-derived CTL epitope recognised by HLA-A2-restricted CD8⁺ T cells

Abstract.

Microsatellite instability (MSI) is recognised as genome-wide alterations in repetitive DNA sequences caused by defects in the DNA mismatch repair machinery. Such mutation patterns have been found in almost all analysed malignancies from patients with hereditary non-polyposis colorectal cancer, and in approximately 15% of sporadic colorectal cancers. In cancers with the MSI phenotype, microsatellite-like sequences in coding regions of various cancer-related genes, including transforming growth factor β receptor type II (TGFβRII), are targets for mutations. The TGFβRII gene harbours a 10-bp polyadenine tract, and mutations within this region are found in 90% of colorectal cancers with MSI. The frameshift mutations result in new amino acid sequences in the C-terminal part of the proteins, prematurely terminating where a novel stop codon appears. In this study we have defined a new cytotoxic T lymphocyte (CTL) epitope (RLSSCVPVA ), carrying a good HLA-A*0201 binding motif, and resulting from the most common frameshift mutation in TGFβRII. A CTL line and several CTL clones were generated from an HLA-A2⁺ normal donor by repeated stimulation of T cells with dendritic cells pulsed with the peptide. One of the CTL clones was able to kill an HLA-A2⁺ colon cancer cell line harbouring mutant TGFβRII. This epitope may be a valuable component in cancer vaccines directed at MSI-positive carcinomas.