Construction and molecular characterization of a T-cell receptor-like antibody and CAR-T cells specific for minor histocompatibility antigen HA-1H.
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.
Department of Hematology, School of Medicine, Fujita Health University, Toyoake, Japan.
Y
Y
Inaguma
Division of Antibody Project, Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Japan.
Y
Y
Akahori
Department of Hematology, School of Medicine, Fujita Health University, Toyoake, Japan.
Y
Y
Murayama
Division of Immunology, Aichi Cancer Center Research Center, Nagoya, Japan.
K
K
Shiraishi
Department of Hematology, School of Medicine, Fujita Health University, Toyoake, Japan.
S
S
Tsuzuki-Iba
Department of Hematology, School of Medicine, Fujita Health University, Toyoake, Japan.
A
A
Endoh
Department of Hematology, School of Medicine, Fujita Health University, Toyoake, Japan.
J
J
Tsujikawa
Division of Immunology, Aichi Cancer Center Research Center, Nagoya, Japan.
A
A
Demachi-Okamura
Division of Immunology, Aichi Cancer Center Research Center, Nagoya, Japan.
K
K
Hiramatsu
HLA Foundation Laboratory, Kyoto, Japan.
H
H
Saji
Department of Hematology, School of Medicine, Fujita Health University, Toyoake, Japan.
Y
Y
Yamamoto
Laboratory of Molecular Biology and Histochemistry, Fujita Health University Joint Research Laboratory, Toyoake, Japan.
N
N
Yamamoto
Department of Immunogenetics, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan.
Y
Y
Nishimura
Division of Immunology, Aichi Cancer Center Research Center, Nagoya, Japan.
T
T
Takahashi
Division of Immunology, Aichi Cancer Center Research Center, Nagoya, Japan.
K
K
Kuzushima
Department of Hematology, School of Medicine, Fujita Health University, Toyoake, Japan.
N
N
Emi
1] Department of Hematology, School of Medicine, Fujita Health University, Toyoake, Japan [2] Division of Immunology, Aichi Cancer Center Research Center, Nagoya, Japan.
Y
Y
Akatsuka
D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology; D000918:Antibody Specificity; D001483:Base Sequence; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D016827:CD8 Antigens / Q000378:metabolism; D000939:Epitopes / Q000276:immunology; D015789:HLA-A2 Antigen / Q000276:immunology*; D006801:Humans; D018345:Mice, Knockout; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D011948:Receptors, Antigen, T-Cell / Q000276:immunology*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology*; D057127:Single-Chain Antibodies / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000502:physiology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D001613:beta 2-Microglobulin / Q000235:genetics
D000911:Antibodies, Monoclonal; D016827:CD8 Antigens; D000939:Epitopes; D015789:HLA-A2 Antigen; D011948:Receptors, Antigen, T-Cell; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies; D001613:beta 2-Microglobulin