Because of tolerance mechanisms, it has been hard to identify the T cell receptors (TCRs) of high-avidity T cells against self (for example, tumor) antigens. TCRs that are specific for foreign human antigens from the nontolerant T cell repertoire can be identified in mice. Moreover, if mice are constructed to express the human TCR repertoire, they can be used to analyze the unskewed repertoire against human self antigens. Here we generated transgenic mice with the entire human TCRαβ gene loci (1.1 and 0.7 Mb), whose T cells express a diverse human TCR repertoire that compensates for mouse TCR deficiency. A human major histocompatibility class I transgene increases the generation of CD8+ T cells with human compared to mouse TCRs. Functional CD8+ T cells against several human tumor antigens were induced, and those against the Melan-A melanoma antigen used similar TCRs to those that have been detected in T cell clones from individuals with autoimmune vitiligo or melanoma. These mice will allow researchers to identify pathogenic and therapeutic human TCRs.
Access optionsAccess options
Subscribe to Journal
Get full journal access for 1 year
only $18.75 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Hedrick, S.M., Cohen, D.I., Nielsen, E.A. & Davis, M.M. Isolation of cDNA clones encoding T cell-specific membrane-associated proteins. Nature 308, 149–153 (1984).
Yanagi, Y. et al. A human T cell–specific cDNA clone encodes a protein having extensive homology to immunoglobulin chains. Nature 308, 145–149 (1984).
Chien, Y.H., Gascoigne, N.R.J., Kavaler, J., Lee, N.E. & Davis, M.M. Somatic recombination in a murine T cell receptor gene. Nature 309, 322–326 (1984).
Kisielow, P., Teh, H.S., Bluthmann, H. & von Boehmer, H. Positive selection of antigen-specific T cells in thymus by restricting MHC molecules. Nature 335, 730–733 (1988).
Sha, W.C. et al. Positive and negative selection of an antigen receptor on T cells in transgenic mice. Nature 336, 73–76 (1988).
Jameson, S.C., Hogquist, K.A. & Bevan, M.J. Positive selection of thymocytes. Annu. Rev. Immunol. 13, 93–126 (1995).
Tanchot, C., Lemonnier, F.A., Pérarnau, B., Freitas, A.A. & Rocha, B. Differential requirements for survival and proliferation of CD8 naive or memory T cells. Science 276, 2057–2062 (1997).
Zinkernagel, R.M. & Doherty, P.C. Restriction of in vitro T cell-mediated cytotoxicity in lymphocytic choriomeningitis within a syngeneic or semiallogeneic system. Nature 248, 701–702 (1974).
Zinkernagel, R.M. et al. On the thymus in the differentiation of “H-2 self-recognition” by T cells: evidence for dual recognition? J. Exp. Med. 147, 882–896 (1978).
Blackman, M. et al. The T cell repertoire may be biased in favor of MHC recognition. Cell 47, 349–357 (1986).
Zerrahn, J., Held, W. & Raulet, D.H. The MHC reactivity of the T cell repertoire prior to positive and negative selection. Cell 88, 627–636 (1997).
Marrack, P., Scott-Browne, J.P., Dai, S., Gapin, L. & Kappler, J.W. Evolutionarily conserved amino acids that control TCR-MHC interaction. Annu. Rev. Immunol. 26, 171–203 (2008).
Scott-Browne, J.P., White, J., Kappler, J.W., Gapin, L. & Marrack, P. Germline-encoded amino acids in the αβ T cell receptor control thymic selection. Nature 458, 1043–1046 (2009).
Kreslavsky, T., Gleimer, M. & von Boehmer, H. αβ versus γδ lineage choice at the first TCR-controlled checkpoint. Curr. Opin. Immunol. 22, 185–192 (2010).
Sensi, M. et al. Cytotoxic T lymphocyte clones from different patients display limited T-cell-receptor variable-region gene usage in HLA-A2–restricted recognition of the melanoma antigen Melan-A/MART-1. Proc. Natl. Acad. Sci. USA 92, 5674–5678 (1995).
Dietrich, P.-Y. et al. Melanoma patients respond to a cytotoxic T lymphocyte–defined self-peptide with diverse and nonoverlapping T-cell receptor repertoires. Cancer Res. 61, 2047–2054 (2001).
Mantovani, S. et al. Dominant TCR-α requirements for a self antigen recognition in humans. J. Immunol. 169, 6253–6260 (2002).
Trautmann, L. et al. Dominant TCR Vα usage by virus and tumor-reactive T cells with wide affinity ranges for their specific antigens. Eur. J. Immunol. 32, 3181–3190 (2002).
Mantovani, S. et al. Molecular and functional bases of self-antigen recognition in long-term persistent melanocyte-specific CD8+ T cells in one vitiligo patient. J. Invest. Dermatol. 121, 308–314 (2003).
Sherman, L.A. et al. Strategies for tumor elimination by cytotoxic T lymphocytes. Crit. Rev. Immunol. 18, 47–54 (1998).
Schumacher, T.N. T-cell-receptor gene therapy. Nat. Rev. Immunol. 2, 512–519 (2002).
Pascolo, S. et al. HLA-A2.1-restricted education and cytolytic activity of CD8+ T lymphocytes from β2 microglobulin (β2m) HLA-A2.1 monochain transgenic H-2Db β2m double knockout mice. J. Exp. Med. 185, 2043–2051 (1997).
Mombaerts, P. et al. Mutations in T cell antigen receptor genes α and β block thymocyte development at different stages. Nature 360, 225–231 (1992).
Dudley, E.C., Girardi, M., Owen, M.J. & Hayday, A.C. αβ and γ∂ T cells can share a late common precursor. Curr. Biol. 5, 659–669 (1995).
Kang, J., Volkmann, A. & Raulet, D.H. Evidence that γδ versus αβ T cell fate determination is initiated independently of T cell signaling. J. Exp. Med. 193, 689–698 (2001).
Joachims, M.L., Chain, J.L., Hooker, S.W., Knott-Craig, C.J. & Thompson, L.F. Human αβ and γδ thymocyte development: TCR gene rearrangements, intracellular TCRβ expression and γδ developmental potential—differences between men and mice. J. Immunol. 176, 1543–1552 (2006).
Kreslavsky, T., Garbe, A.I., Krueger, A. & von Boehmer, H. T cell receptor–instructed αβ versus γδ lineage commitment revealed by single-cell analysis. J. Exp. Med. 205, 1173–1186 (2008).
Burtrum, D.B., Kim, S., Dudley, E.C., Hayday, A.C. & Petrie, H.T. TCR gene recombination and αβ-γδ lineage divergence: productive TCR-β rearrangement is neither exclusive nor preclusive of γδ cell development. J. Immunol. 157, 4293–4296 (1996).
Almeida, A.R.M., Borghans, J.A.M. & Freitas, A.A. T cell homeostasis: Thymus regeneration and peripheral T cell restoration in mice with a reduced fraction of competent precursors. J. Exp. Med. 194, 591–599 (2001).
Lonberg, N. et al. Antigen-specific human antibodies from mice comprising four distinct genetic modifications. Nature 368, 856–859 (1994).
Arstila, T.P. et al. A direct estimate of the human αβ T cell receptor diversity. Science 286, 958–961 (1999).
Casrouge, A. et al. Size estimate of the αβ TCR repertoire of naïve mouse splenocytes. J. Immunol. 164, 5782–5787 (2000).
Alexander, J. et al. Derivation of HLA-A11/Kb transgenic mice: functional CTL repertoire and recognition of human A11-restricted CTL epitopes. J. Immunol. 159, 4753–4761 (1997).
Markie, D. (ed.) YAC Protocols. Meth. Mol. Biol. 54 (Humana, 1995).
Engels, B. et al. Retroviral vectors for high-level transgene expression in T lymphocytes. Hum. Gene Ther. 14, 1155–1168 (2003).
We thank M. Textor, M. Rösch, A. Gärtner and C. Westen for technical assistance; A. Garratt, E. Rohde and A. Samuels for help with ES cell culture; M. Hafner and M. Ebel for ES cell injection; H.-P. Rahn for FACS sorting; M. Gong for help with spectratyping and statistical analysis; F. Lemonier (Institut Pasteur) for HHDII mice; E. Mcintyre (Hôpital Necker) and E. Green (US National Institutes of Health) for YACs; and D. Schendel and D. Busch for critically reading the manuscript. This work was supported by grants from the Deutsche Forschungsgemeinschaft (Sonderforschungsbereich TR36) and the Helmholtz-Gemeinschaft Deutscher Forschungszentren (HA-202). W.M. was supported by the European Commission program MUGEN LSHG-CT-2005-005203 at the Helmholtz Centre for Infection Research, Braunschweig, Germany.
The authors declare no competing financial interests.
About this article
Cancer immunotherapy with lymphocytes genetically engineered with T cell receptors for solid cancers
Immunology Letters (2019)
Journal of Cellular Biochemistry (2019)
Advanced Drug Delivery Reviews (2019)
In-vitro blockade of the CD4 receptor co-signal in antigen-specific T-cell stimulation cultures induces the outgrowth of potent CD4 independent T-cell effectors
Journal of Immunological Methods (2018)