ScFv为基础的嵌合抗原受体诱导T细胞攻击IL13Rα2阳性神经胶...
Characterization and Functional Analysis of scFv-based Chimeric Antigen Receptors to Redirect T Cells to IL13Rα2-positive Glioma AbstractImmunotherapy with T cells expressing chimeric antigen receptors (CARs) is an attractive approach to improve outcomes for patients with glioblastoma (GBM). IL13Rα2 is expressed at a high frequency in GBM but not in normal brain, making it a promising CAR T-cell therapy target. IL13Rα2-specific CARs generated up to date contain mutated forms of IL13 as an antigen-binding domain. While these CARs target IL13Rα2, they also recognize IL13Rα1, which is broadly expressed. To overcome this limitation, we constructed a panel of IL13Rα2-specific CARs that contain the IL13Rα2-specific single-chain variable fragment (scFv) 47 as an antigen binding domain, short or long spacer regions, a transmembrane domain, and endodomains derived from costimulatory molecules and CD3.ζ (IL13Rα2-CARs). IL13Rα2-CAR T cells recognized IL13Rα2-positive target cells in coculture and cytotoxicity assays with no cross-reactivity to IL13Rα1. However, only IL13Rα2-CAR T cells with a short spacer region produced IL2 in an antigen-dependent fashion. In vivo, T cells expressing IL13Rα2-CARs with short spacer regions and CD28.ζ, 41BB.ζ, and CD28.OX40.ζ endodomains had potent anti-glioma activity conferring a significant survival advantage in comparison to mice that received control T cells. Thus, IL13Rα2-CAR T cells hold the promise to improve current IL13Rα2-targeted immunotherapy approaches for GBM and other IL13Rα2-positive malignancies. Molecular Therapy (2015);ScFv为基础的嵌合抗原受体诱导T细胞攻击IL13Rα2阳性神经胶质瘤的特征和功能分析摘要:嵌合抗原受体修饰的T细胞免疫治疗对于神经胶质母细胞瘤病人而言，是一个非常吸引人的治疗途径。IL13Rα2分子在胶质母细胞瘤细胞表面高度表达，但是在正常脑细胞表面不怎么表达，从而成为一个理想的CAR-T细胞针对的靶点。IL13Rα2修饰的CARs到目前为止的设计包含变异形式的IL13作为一个抗原结合区域。当这些CARs靶向IL13Rα2时，它们同样靶向广泛表达的IL13Rα1分子。为了克服这样的限制，我们设计了一个特异性的IL13Rα2 CARs，它包含了IL13Rα2特异性的单链可变区域（scFv）47作为一个抗原结合区域，或长或短的间隔区、一个跨膜结构域、来自共刺激分子和CD3的内部区域。在共培养和细胞毒性试验中，IL13Rα2 CAR-T细胞识别IL13Rα2阳性细胞，与IL13Rα1没有交叉反应。然而，只有IL13Rα2-CAR 短间隔区的T细胞产生抗原依赖性。在体内，具有更短间隔区和CD28.ζ、41BB.ζ、CD28、OX40.ζ内域的IL13Rα2-CARsT细胞具有很强的抗胶质瘤活性。因此，IL13Rα2-CAR T细胞有希望改进目前IL13Rα2靶向免疫治疗神经胶质目细胞瘤和其他IL13Rα2阳性肿瘤。出自爱康得
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