Identifying T-cell receptors (TCRs) that bind tumor-associated antigens (TAAs) with optimal affinity is a key bottleneck in the development of adoptive T-cell therapy of cancer1. TAAs are unmutated self proteins, and T cells bearing high-affinity TCRs specific for such antigens are commonly deleted in the thymus2. To identify optimal-affinity TCRs, we generated antigen-negative humanized mice with a diverse human TCR repertoire restricted to the human leukocyte antigen (HLA) A*02:01 (ref. 3). These mice were immunized with human TAAs, for which they are not tolerant, allowing induction of CD8+ T cells with optimal-affinity TCRs. We isolate TCRs specific for the cancer/testis (CT) antigen MAGE-A1 (ref. 4) and show that two of them have an anti-tumor effect in vivo. By comparison, human-derived TCRs have lower affinity and do not mediate substantial therapeutic effects. We also identify optimal-affinity TCRs specific for the CT antigen NY-ESO. Our humanized mouse model provides a useful tool for the generation of optimal-affinity TCRs for T-cell therapy.
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June, C.H. Adoptive T cell therapy for cancer in the clinic. J. Clin. Invest. 117, 1466–1476 (2007).
Theobald, M. et al. Tolerance to p53 by A2.1-restricted cytotoxic T lymphocytes. J. Exp. Med. 185, 833–841 (1997).
Li, L.-P. et al. Transgenic mice with a diverse human T cell antigen receptor repertoire. Nat. Med. 16, 1029–1034 (2010).
van der Bruggen, P. et al. A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma. Science 254, 1643–1647 (1991).
Holler, P.D., Chlewicki, L.K. & Kranz, D.M. TCRs with high affinity for foreign pMHC show self-reactivity. Nat. Immunol. 4, 55–62 (2003).
Linette, G.P. et al. Cardiovascular toxicity and titin cross-reactivity of affinity-enhanced T cells in myeloma and melanoma. Blood 122, 863–871 (2013).
Engels, B., Chervin, A.S., Sant, A.J., Kranz, D.M. & Schreiber, H. Long-term persistence of CD4(+) but rapid disappearance of CD8(+) T cells expressing an MHC class I-restricted TCR of nanomolar affinity. Mol. Ther. 20, 652–660 (2012).
Sadovnikova, E. & Stauss, H.J. Peptide-specific cytotoxic T lymphocytes restricted by nonself major histocompatibility complex class I molecules: reagents for tumor immunotherapy. Proc. Natl. Acad. Sci. USA 93, 13114–13118 (1996).
Johnson, L.A. et al. Gene therapy with human and mouse T-cell receptors mediates cancer regression and targets normal tissues expressing cognate antigen. Blood 114, 535–546 (2009).
Parkhurst, M.R. et al. Characterization of genetically modified T-cell receptors that recognize the CEA:691–699 peptide in the context of HLA-A2.1 on human colorectal cancer cells. Clin. Cancer Res. 15, 169–180 (2009).
Chinnasamy, N. et al. A TCR targeting the HLA-A*0201-restricted epitope of MAGE-A3 recognizes multiple epitopes of the MAGE-A antigen superfamily in several types of cancer. J. Immunol. 186, 685–696 (2011).
Davis, J.L. et al. Development of human anti-murine T-cell receptor antibodies in both responding and nonresponding patients enrolled in TCR gene therapy trials. Clin. Cancer Res. 16, 5852–5861 (2010).
Offringa, R. Antigen choice in adoptive T-cell therapy of cancer. Curr. Opin. Immunol. 21, 190–199 (2009).
Pascolo, S. et al. A MAGE-A1 HLA-A A*0201 epitope identified by mass spectrometry. Cancer Res. 61, 4072–4077 (2001).
Toso, J.F. et al. MAGE-1-specific precursor cytotoxic T-lymphocytes present among tumor-infiltrating lymphocytes from a patient with breast cancer: characterization and antigen-specific activation. Cancer Res. 56, 16–20 (1996).
Ottaviani, S., Zhang, Y., Boon, T. & van der Bruggen, P.A. MAGE-1 antigenic peptide recognized by human cytolytic T lymphocytes on HLA-A2 tumor cells. Cancer Immunol. Immunother. 54, 1214–1220 (2005).
Huijbers, I.J. et al. Minimal tolerance to a tumor antigen encoded by a cancer-germline gene. J. Immunol. 188, 111–121 (2012).
Lee, E.-C. et al. Complete humanization of the mouse immunoglobulin loci enables efficient therapeutic antibody discovery. Nat. Biotechnol. 32, 356–363 (2014).
Sommermeyer, D. & Uckert, W. Minimal amino acid exchange in human TCR constant regions fosters improved function of TCR gene-modified T cells. J. Immunol. 184, 6223–6231 (2010).
Morgan, R.A. et al. Cancer regression and neurological toxicity following anti-MAGE-A3 TCR gene therapy. J. Immunother. 36, 133–151 (2013).
Linnemann, C. et al. High-throughput identification of antigen-specific TCRs by TCR gene capture. Nat. Med. 19, 1534–1541 (2013).
Chen, J.-L. et al. Structural and kinetic basis for heightened immunogenicity of T cell vaccines. J. Exp. Med. 201, 1243–1255 (2005).
Vierboom, M.P. et al. Tumor eradication by wild-type p53-specific cytotoxic T lymphocytes. J. Exp. Med. 186, 695–704 (1997).
Scott-Browne, J.P., White, J., Kappler, J.W., Gapin, L. & Marrack, P. Germline-encoded amino acids in the alphabeta T-cell receptor control thymic selection. Nature 458, 1043–1046 (2009).
Aleksic, M. et al. Different affinity windows for virus and cancer-specific T-cell receptors: implications for therapeutic strategies. Eur. J. Immunol. 42, 3174–3179 (2012).
Schmid, D.A. et al. Evidence for a TCR affinity threshold delimiting maximal CD8 T cell function. J. Immunol. 184, 4936–4946 (2010).
Wang, B. et al. A single peptide-MHC complex positively selects a diverse and specific CD8 T cell repertoire. Science 326, 871–874 (2009).
Birnbaum, M.E. et al. Deconstructing the peptide-MHC specificity of T cell recognition. Cell 157, 1073–1087 (2014).
Gotter, J., Brors, B., Hergenhahn, M. & Kyewski, B. Medullary epithelial cells of the human thymus express a highly diverse selection of tissue-specific genes colocalized in chromosomal clusters. J. Exp. Med. 199, 155–166 (2004).
Kammertoens, T. & Blankenstein, T. It's the peptide-MHC affinity, stupid. Cancer Cell 23, 429–431 (2013).
Huang, L.Q. et al. Cytolytic T lymphocytes recognize an antigen encoded by MAGE-A10 on a human melanoma. J. Immunol. 162, 6849–6854 (1999).
Zarour, H. et al. The majority of autologous cytolytic T-lymphocyte clones derived from peripheral blood lymphocytes of a melanoma patient recognize an antigenic peptide derived from Gene Pmel17/gp100. J. Invest. Dermatol. 107, 63–67 (1996).
Hérin, M. et al. Production of stable cytolytic T-cell clones directed against autologous human melanoma. Int. J. Cancer 39, 390–396 (1987).
Weynants, P. et al. Derivation of tumor-specific cytolytic T-cell clones from two lung cancer patients with long survival. Am. J. Respir. Crit. Care Med. 159, 55–62 (1999).
Pellat-Deceunynck, C. et al. The cancer germ-line genes MAGE-1, MAGE-3 and PRAME are commonly expressed by human myeloma cells. Eur. J. Immunol. 30, 803–809 (2000).
Bredenbeck, A. et al. Coordinated expression of clustered cancer/testis genes encoded in a large inverted repeat DNA structure. Gene 415, 68–73 (2008).
Sommermeyer, D. et al. NY-ESO-1 antigen-reactive T cell receptors exhibit diverse therapeutic capability. Int. J. Cancer 132, 1360–1367 (2013).
Morita, S., Kojima, T. & Kitamura, T. Plat-E: an efficient and stable system for transient packaging of retroviruses. Gene Ther. 7, 1063–1066 (2000).
Leisegang, M. et al. Enhanced functionality of T cell receptor-redirected T cells is defined by the transgene cassette. J. Mol. Med. (Berl) 86, 573–583 (2008).
Kuball, J. et al. Facilitating matched pairing and expression of TCR chains introduced into human T cells. Blood 109, 2331–2338 (2007).
Cohen, C.J. et al. Enhanced antitumor activity of T cells engineered to express T-cell receptors with a second disulfide bond. Cancer Res. 67, 3898–3903 (2007).
Engels, B. et al. Retroviral vectors for high-level transgene expression in T lymphocytes. Hum. Gene Ther. 14, 1155–1168 (2003).
Uckert, W. et al. Efficient gene transfer into primary human CD8+ T lymphocytes by MuLV-10A1 retrovirus pseudotype. Hum. Gene Ther. 11, 1005–1014 (2000).
Heemskerk, M.H.M. et al. Redirection of antileukemic reactivity of peripheral T lymphocytes using gene transfer of minor histocompatibility antigen HA-2-specific T-cell receptor complexes expressing a conserved alpha joining region. Blood 102, 3530–3540 (2003).
Wilde, S. et al. Dendritic cells pulsed with RNA encoding allogeneic MHC and antigen induce T cells with superior antitumor activity and higher TCR functional avidity. Blood 114, 2131–2139 (2009).
de Castro, E. et al. ScanProsite: detection of PROSITE signature matches and ProRule-associated functional and structural residues in proteins. Nucleic Acids Res. 34, W362–W365 (2006).
The authors thank G. Willimsky for discussion, S. Kupsch and S. Fürl for technical assistance, and I. Hoeft for animal caretaking. C. Linnemann and T. Schumacher kindly provided the NY-ESO-1157/HLA-A2 multimer. This work was supported by the Deutsche Forschungsgemeinschaft (Sonderforschungsbereich TR36) and the Berlin Institute of Health (BIH).
The Max-Delbrück-Center for Molecular Medicine (T.B., M.O., C.L.) applied for a patent on the MAGE-A1-specific TCRs.
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Obenaus, M., Leitão, C., Leisegang, M. et al. Identification of human T-cell receptors with optimal affinity to cancer antigens using antigen-negative humanized mice. Nat Biotechnol 33, 402–407 (2015). https://doi.org/10.1038/nbt.3147
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