Construction and molecular characterization of a T-cell receptor-like antibody and CAR-T cells specific for minor histocompatibility antigen HA-1H
Construction and molecular characterization of a T-cell receptor-like antibody and CAR-T cells specific for minor histocompatibility antigen HA-1H
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DOI：
10.1038/gt.2014.30
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发表时间：
2014-06-01
期刊：
影响因子：
5.1
通讯作者：
Akatsuka, Y.
中科院分区：
文献类型：
--
作者：
Inaguma, Y.;Akahori, Y.;Akatsuka, Y.
针对靶抗原的 T 细胞受体 (TCR) 基因转移到 T 淋巴细胞中，已被用于有效生成抗肿瘤 T 细胞，而无需体外诱导和扩增具有同源特异性的 T 细胞。或者，T 细胞已用 TCR 样抗体或嵌合抗原受体 (CAR) 进行基因修饰。我们发现，用掺入HA-1 H次要组织相容性抗原（mHag）肽和β2-微球蛋白（HA-1 H/HLA-A2）的四聚化重组HLA-A2免疫HLA-A2转基因小鼠可产生高度特异性的抗体。一种单链可变区部分 (scFv) 抗体 #131 对 HA-1 H/HLA-A2 复合物表现出高亲和力 (K-D=14.9 nm)。用与 CD28 跨膜和 CD3 zeta 结构域偶联的 #131 scFV 转导的原代人 T 细胞被 HA-1 H/HLA-A2 四聚体染色，其染色强度略强于内源性 HLA-A2 和 HA-1 H 阳性细胞特异性的细胞毒性 T 淋巴细胞 (CTL) 克隆。尽管与同源 CTL 克隆相比，#131 scFv CAR-T 细胞需要高 100 倍以上的抗原密度才能发挥细胞毒性，但它们可以针对表达 HLA-A2 和 HA-1 H 转基因的细胞产生炎症细胞因子。这些数据表明，具有高亲和力抗原受体的 T 细胞降低了用低密度肽/MHC 复合物（类似于每个细胞 100 个）裂解靶标的能力，尽管它们可以在细胞因子产生水平上做出反应。
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 beta 2-microglobulin (HA-1 H/HLA-A2) generate highly specific antibodies. One single-chain variable region moiety (scFv) antibody, #131, demonstrated high affinity (K-D=14.9 nm) for the HA-1 H/HLA-A2 complex. Primary human T cells transduced with #131 scFV coupled to CD28 transmembrane and CD3 zeta 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 (similar to 100 per cell), while they could respond, at cytokine production level.