Reprogramming CD19-specific T cells with IL-21 signaling can improve adoptive immunotherapy of B-lineage malignancies.
Reprogramming CD19-specific T cells with IL-21 signaling can improve adoptive immunotherapy of B-lineage malignancies.
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DOI：
10.1158/0008-5472.can-10-3843
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发表时间：
2011-05-15
期刊：
影响因子：
11.2
通讯作者：
Cooper LJ
中科院分区：
文献类型：
--
作者：
Singh H;Figliola MJ;Dawson MJ;Huls H;Olivares S;Switzer K;Mi T;Maiti S;Kebriaei P;Lee DA;Champlin RE;Cooper LJ
提高表达嵌合抗原受体(CAR)的T细胞的治疗效果是控制B细胞恶性肿瘤的一个重要目标。最近，一种内在的策略已经被开发出来，通过修改CAR本身来改善T细胞信号。在这里，我们报告了第二种外在方法，基于改变培养环境，以数字方式扩增具有所需表型的CAR+T细胞。因为，在组织培养中加入IL-21可以改善CAR依赖的T细胞效应器功能。我们利用电转移睡美人系统引入CAR转座子，并在CD19+人工抗原提呈细胞(AAPC)上选择性地扩增CAR+T细胞。当IL-21存在时，CD19特异性T细胞优先数扩增，其裂解并产生干扰素-γ以响应CD19。这些数量扩增的CAR+T细胞群体表现出早期的记忆表面表型，其特征是CD62L+CD28+，以及天真T细胞的转录特征。相反，仅用外源性IL-2培养的T细胞往往会导致CD19特异性的CD4+T细胞过度生长。此外，用IL-21培养的CAR+T细胞过继转移可改善对小鼠CD19+B细胞恶性的控制。为了提供协同信号来繁殖CAR+T细胞，我们开发了一种新的IL-21突变体，它结合到AAPC的细胞表面，取代了对可溶性IL-21的需要。我们的发现表明，IL-21可以提供外在的重编程信号，以产生所需的CAR+T细胞，用于有效的免疫治疗。
Improving the therapeutic efficacy of T cells expressing a chimeric antigen receptor (CAR) represents an important goal in efforts to control B-cell malignancies. Recently an intrinsic strategy has been developed to modify the CAR itself to improve T-cell signaling. Here we report a second extrinsic approach based on altering the culture milieu to numerically expand CAR+ T cells with a desired phenotype. For, the addition of IL-21 to tissue culture improves CAR-dependent T-cell effector functions. We used electrotransfer of Sleeping Beauty (SB) system to introduce a CAR transposon and selectively propagate CAR+ T cells on CD19+ artificial antigen-presenting cells (aAPC). When IL-21 was present, there was preferential numeric expansion of CD19-specific T cells which lysed and produced IFN-γ in response to CD19. Populations of these numerically expanded CAR+ T cells displayed an early memory surface phenotype characterized as CD62L+CD28+ and a transcriptional profile of naïve T cells. In contrast, T cells propagated with only exogenous IL-2 tended to result in an overgrowth of CD19-specific CD4+ T cells. Furthermore, adoptive transfer of CAR+ T cells cultured with IL-21 exhibited improved control of CD19+ B-cell malignancy in mice. To provide coordinated signaling to propagate CAR+ T cells, we developed a novel mutein of IL-21 bound to the cell surface of aAPC that replaced the need for soluble IL-21. Our findings demonstrate that IL-21 can provide an extrinsic reprogramming signal to generate desired CAR+ T cells for effective immunotherapy.