Generating De Novo Antigen-specific Human T Cell Receptors by Retroviral Transduction of Centric Hemichain.
T cell receptors (TCRs) are used clinically to direct the specificity of T cells to target tumors as a promising modality of immunotherapy. Therefore, cloning TCRs specific for various tumor-associated antigens has been the goal of many studies. To elicit an effective T cell response, the TCR must recognize the target antigen with optimal affinity. However, cloning such TCRs has been a challenge and many available TCRs possess sub-optimal affinity for the cognate antigen. In this protocol, we describe a method of cloning de novo high affinity antigen-specific TCRs using existing TCRs by exploiting hemichain centricity. It is known that for some TCRs, each TCRα or TCRβ hemichain do not contribute equally to antigen recognition, and the dominant hemichain is referred to as the centric hemichain. We have shown that by pairing the centric hemichain with counter-chains differing from the original counter-chain, we are able to maintain the antigen specificity, while modulating its interaction strength for the cognate antigen. Thus, the therapeutic potential of a given TCR can be improved by optimizing the pairing between the centric and counter hemichains.
Department of Immunology, University of Toronto; Princess Margaret Cancer Centre, University Health Network.
Tingxi
T
Guo
Princess Margaret Cancer Centre, University Health Network.
Toshiki
T
Ochi
Princess Margaret Cancer Centre, University Health Network.
Munehide
M
Nakatsugawa
Princess Margaret Cancer Centre, University Health Network.
Yuki
Y
Kagoya
Department of Immunology, University of Toronto; Princess Margaret Cancer Centre, University Health Network.
Mark
M
Anczurowski
Department of Immunology, University of Toronto; Princess Margaret Cancer Centre, University Health Network.
Chung-Hsi
CH
Wang
Princess Margaret Cancer Centre, University Health Network.
Muhammed A
MA
Rahman
Princess Margaret Cancer Centre, University Health Network.
Kayoko
K
Saso
Department of Immunology, University of Toronto; Princess Margaret Cancer Centre, University Health Network.
Marcus O
MO
Butler
Department of Immunology, University of Toronto; Princess Margaret Cancer Centre, University Health Network; naoto.hirano@utoronto.ca.
Naoto
N
Hirano
D000951:Antigens, Neoplasm; D000939:Epitopes; D006801:Humans; D007167:Immunotherapy; D011948:Receptors, Antigen, T-Cell; D012190:Retroviridae; D013601:T-Lymphocytes
D000951:Antigens, Neoplasm; D000939:Epitopes; D011948:Receptors, Antigen, T-Cell