Systematic testing and specificity mapping of alloantigen-specific chimeric antigen receptors in regulatory T cells
Systematic testing and specificity mapping of alloantigen-specific chimeric antigen receptors in regulatory T cells
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DOI：
10.1172/jci.insight.123672
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发表时间：
2019-03-21
期刊：
影响因子：
8
通讯作者：
Levings, Megan K.
中科院分区：
文献类型：
--
作者：
Dawson, Nicholas A. J.;Lamarche, Caroline;Levings, Megan K.
嵌合抗原受体（CAR）技术可用于工程化调节性T细胞（Tcells）的抗原特异性，并提高其在多种疾病模型中作为过继性细胞治疗的效力。作为合成受体，汽车具有免疫原性的风险，特别是当衍生自非人抗体时。使用编码源自小鼠抗体的单链可变片段（Fv）的HLA-A*02：01特异性CAR（A2-CAR），我们开发了一组20个人源化A2-汽车（hA 2-汽车）。系统测试证明了表达的变化，以及结合HLA-A*02：01和刺激体外人Treg抑制的能力。此外，我们开发了一种新的方法来全面绘制汽车的同种异体抗原特异性，揭示了人源化降低了HLA-A交叉反应性。体内生物发光成像显示hA 2-CAR T细胞在表达A2的同种异体移植物中的快速运输和持久性，最终迁移到引流淋巴结。连续转移hA 2-CAR T细胞可抑制HLA-A2（+）细胞介导的异种移植物抗宿主病，并减少人类HLA-A2（+）皮肤同种异体移植物的排斥反应。这些数据提供了用于系统开发和特异性测试人源化同种异体抗原特异性汽车的平台，所述CAR可用于工程化特异性和治疗性T细胞的归巢。
Chimeric antigen receptor (CAR) technology can be used to engineer the antigen specificity of regulatory T cells (Tregs) and improve their potency as an adoptive cell therapy in multiple disease models. As synthetic receptors, CARs carry the risk of immunogenicity, particularly when derived from nonhuman antibodies. Using an HLA-A*02:01-specific CAR (A2-CAR) encoding a single-chain variable fragment (Fv) derived from a mouse antibody, we developed a panel of 20 humanized A2-CARs (hA2-CARs). Systematic testing demonstrated variations in expression, and ability to bind HLA-A*02:01 and stimulate human Treg suppression in vitro. In addition, we developed a new method to comprehensively map the alloantigen specificity of CARs, revealing that humanization reduced HLA-A cross-reactivity. In vivo bioluminescence imaging showed rapid trafficking and persistence of hA2-CAR Tregs in A2-expressing allografts, with eventual migration to draining lymph nodes. Adoptive transfer of hA2-CAR Tregs suppressed HLA-A2(+) cell-mediated xenogeneic graft-versus-host disease and diminished rejection of human HLA-A2(+) skin allografts. These data provide a platform for systematic development and specificity testing of humanized alloantigen-specific CARs that can be used to engineer specificity and homing of therapeutic Tregs.