Linnemann, Carsten2; Heemskerk, Bianca3; Kvistborg, Pia3; Kluin, Roelof J C3; Bolotin, Dmitriy A3; Chen, Xiaojing3; Bresser, Kaspar3; Nieuwland, Marja3; Schotte, Remko3; Michels, Samira3; Gomez-Eerland, Raquel3; Jahn, Lorenz3; Hombrink, Pleun3; Legrand, Nicolas3; Shu, Chengyi Jenny3; Mamedov, Ilgar Z3; Velds, Arno3; Blank, Christian U3; Haanen, John B A G3; Turchaninova, Maria A3; Kerkhoven, Ron M3; Spits, Hergen3; Hadrup, Sine Reker1; Heemskerk, Mirjam H M3; Blankenstein, Thomas3; Chudakov, Dmitriy M3; Bendle, Gavin M3; Schumacher, Ton N M3
1 Department of Haematology, Herlev and Gentofte Hospital, The Capital Region of Denmark2 Division of Immunology, The Netherlands Cancer Institute, Amsterdam, The Netherlands.3 unknown
The transfer of T cell receptor (TCR) genes into patient T cells is a promising approach for the treatment of both viral infections and cancer. Although efficient methods exist to identify antibodies for the treatment of these diseases, comparable strategies to identify TCRs have been lacking. We have developed a high-throughput DNA-based strategy to identify TCR sequences by the capture and sequencing of genomic DNA fragments encoding the TCR genes. We establish the value of this approach by assembling a large library of cancer germline tumor antigen-reactive TCRs. Furthermore, by exploiting the quantitative nature of TCR gene capture, we show the feasibility of identifying antigen-specific TCRs in oligoclonal T cell populations from either human material or TCR-humanized mice. Finally, we demonstrate the ability to identify tumor-reactive TCRs within intratumoral T cell subsets without knowledge of antigen specificities, which may be the first step toward the development of autologous TCR gene therapy to target patient-specific neoantigens in human cancer.
Nature Medicine, 2013, Vol 19, Issue 11, p. 1534-41