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Construction and molecular characterization of a T-cell receptor-like antibody and CAR-T cells specific for minor histocompatibility antigen HA-1H

Gene Therapy volume 21, pages 575–584 (2014)Cite this article

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Abstract

The genetic transfer of T-cell receptors (TCRs) directed toward target antigens into T lymphocytes has been used to generate antitumor T cells efficiently without the need for the in vitro induction and expansion of T cells with cognate specificity. Alternatively, T cells have been gene-modified with a TCR-like antibody or chimeric antigen receptor (CAR). We show that immunization of HLA-A2 transgenic mice with tetramerized recombinant HLA-A2 incorporating HA-1 H minor histocompatibility antigen (mHag) peptides and β2-microglobulin (HA-1 H/HLA-A2) generate highly specific antibodies. One single-chain variable region moiety (scFv) antibody, #131, demonstrated high affinity (KD=14.9 nM) for the HA-1 H/HLA-A2 complex. Primary human T cells transduced with #131 scFV coupled to CD28 transmembrane and CD3ζ domains were stained with HA-1 H/HLA-A2 tetramers slightly more intensely than a cytotoxic T lymphocyte (CTL) clone specific for endogenously HLA-A2- and HA-1 H-positive cells. Although #131 scFv CAR-T cells required >100-fold higher antigen density to exert cytotoxicity compared with the cognate CTL clone, they could produce inflammatory cytokines against cells expressing HLA-A2 and HA-1 H transgenes. These data implicate that T cells with high-affinity antigen receptors reduce the ability to lyse targets with low-density peptide/MHC complexes (~100 per cell), while they could respond at cytokine production level.

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Acknowledgements

We thank Dr W Ho for critically reading the manuscript; Ms Sayoko Ogata and Ms Hiromi Tamaki for their technical expertise. This study was supported in part by Grant-in-Aid for Scientific Research (C)(24591435), from the Ministry of Education, Culture, Science, Sports and Technology, Japan; Grants for Research on the Human Genome, Tissue Engineering Food Biotechnology and the Second and Third Team Comprehensive 10-year Strategy for Cancer Control, from the Ministry of Health, Labour and Welfare, Japan; and a grant from the Japan Leukemia Research Fund (2013). This study was supported in part by Grant-in-Aid for Scientific Research (C)(24591435), from the Ministry of Education, Culture, Science, Sports and Technology, Japan; Grants for Research on the Human Genome, Tissue Engineering Food Biotechnology and the Second and Third Team Comprehensive 10-year Strategy for Cancer Control, from the Ministry of Health, Labour and Welfare, Japan; and a grant from the Japan Leukemia Research Fund (2013).

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Authors and Affiliations

  1. Department of Hematology, School of Medicine, Fujita Health University, Toyoake, Japan

    Y Inaguma, Y Murayama, S Tsuzuki-Iba, A Endoh, J Tsujikawa, Y Yamamoto, N Emi & Y Akatsuka

  2. Division of Antibody Project, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan

    Y Akahori

  3. Division of Immunology, Aichi Cancer Center Research Center, Nagoya, Japan

    K Shiraishi, A Demachi-Okamura, K Hiramatsu, T Takahashi, K Kuzushima & Y Akatsuka

  4. HLA Foundation Laboratory, Kyoto, Japan

    H Saji

  5. Laboratory of Molecular Biology and Histochemistry, Fujita Health University Joint Research Laboratory, Toyoake, Japan

    N Yamamoto

  6. Department of Immunogenetics, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan

    Y Nishimura

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Correspondence to Y Akatsuka.

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Inaguma, Y., Akahori, Y., Murayama, Y. et al. Construction and molecular characterization of a T-cell receptor-like antibody and CAR-T cells specific for minor histocompatibility antigen HA-1H. Gene Ther 21, 575–584 (2014). https://doi.org/10.1038/gt.2014.30

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