A novel TCR-like CAR with specificity for PR1/HLA-A2 effectively targets myeloid leukemia in vitro when expressed in human adult peripheral blood and cord blood T cells.
A novel TCR-like CAR with specificity for PR1/HLA-A2 effectively targets myeloid leukemia in vitro when expressed in human adult peripheral blood and cord blood T cells.
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DOI：
10.1016/j.jcyt.2016.05.001
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发表时间：
2016-08
期刊：
影响因子：
4.5
通讯作者：
Molldrem JJ
中科院分区：
文献类型：
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作者：
Ma Q;Garber HR;Lu S;He H;Tallis E;Ding X;Sergeeva A;Wood MS;Dotti G;Salvado B;Ruisaard K;Clise-Dwyer K;John LS;Rezvani K;Alatrash G;Shpall EJ;Molldrem JJ
PR 1肽来源于白血病相关抗原蛋白酶3和中性粒细胞弹性蛋白酶，在急性髓性白血病（AML）中在HLA-A2上过表达。我们开发了一种T细胞受体（TCR）样单克隆抗体（8 F4），其结合AML细胞表面上的PR 1/HLA-A2复合物，在体外有效地杀死它们并在临床前模型中消除它们。使用对PR 1/HLA-A2表位具有高亲和力的人源化8 F4（h8 F4）来构建h8 F4-嵌合抗原受体（CAR），其被转导到T细胞中以介导抗白血病活性。人T细胞被转导以表达PR 1/HLA-A2特异性CAR（h8 F4-CAR-T细胞），该CAR含有h8 F4的单链抗体，通过CD 28的跨膜和细胞内共刺激结构域与CD 3 zeta链的细胞内信号传导内域融合。成年人正常外周血（PB）T细胞用h8 F4-CAR构建体有效转导，并且在体外主要显示效应记忆表型，具有少量（12%）的中枢记忆细胞。脐带血（UCB）T细胞也可以用h8 F4-CAR有效地转导。PB和UCB衍生的h8 F4-CAR-T细胞特异性识别PR 1/HLA-A2复合物，并能够以HLA-A2依赖性方式杀死白血病细胞系和原代AML母细胞。表达源自TCR样8 F4抗体的CAR的人成人PB和UCB衍生的T细胞在体外快速有效地杀死AML。我们的数据可能会导致AML的新治疗模式，其中靶向白血病干细胞可以转移长期免疫力以防止复发。
The PR1 peptide, derived from the leukemia-associated antigens proteinase 3 and neutrophil elastase, is overexpressed on HLA-A2 in acute myeloid leukemia (AML). We developed a T cell receptor (TCR)-like monoclonal antibody (8F4) that binds the PR1/HLA-A2 complex on the surface of AML cells efficiently killing them in vitro and eliminating them in preclinical models. Humanized 8F4 (h8F4) with high affinity for the PR1/HLA-A2 epitope was used to construct an h8F4- chimeric antigen receptor (CAR) that was transduced into T-cells to mediate anti-leukemia activity. Human T cells were transduced to express the PR1/HLA-A2-specific CAR (h8F4-CAR-T cells) containing the scFv of h8F4 fused to the intracellular signaling endodomain of CD3 zeta chain through the transmembrane and intracellular costimulatory domain of CD28. Adult human normal peripheral blood (PB) T cells were efficiently transduced with the h8F4-CAR construct and predominantly displayed an effector memory phenotype with a minor population (12%) of central memory cells in vitro. Umbilical cord blood (UCB) T cells could also be efficiently transduced with the h8F4-CAR. The PB and UCB-derived h8F4-CAR-T cells specifically recognized the PR1/HLA-A2 complex and were capable of killing leukemia cell lines and primary AML blasts in an HLA-A2-dependent manner. Human adult PB and UCB-derived T cells expressing a CAR derived from the TCR-like 8F4 antibody rapidly and efficiently kill AML in vitro. Our data could lead to a new treatment paradigm for AML in which targeting leukemia stem cells could transfer long-term immunity to protect against relapse.