Therapies that bind with immune cells and redirect their cytotoxic activity toward diseased cells represent a promising and versatile approach to immunotherapy with applications in cancer, lupus, and other diseases; traditional methods for discovering these therapies, however, are often time-intensive and lack the throughput of related target-based discovery approaches. Inspired by the observation that the cytokine, IL-12, can enhance antileukemic activity of the clinically approved T cell redirecting therapy, blinatumomab, here we describe the structure and assembly of a chimeric immune cell-redirecting agent which redirects the lytic activity of primary human T cells toward leukemic B cells and simultaneously cotargets the delivery of T cell-stimulating IL-12. We further describe a novel method for the parallel assembly of compositionally diverse libraries of these bispecific T cell engaging cytokines (BiTEokines) and their high-throughput phenotypic screening, requiring just days for hit identification and the analysis of composition-function relationships. Using this approach, we identified CD19 × CD3 × IL12 compounds that exhibit ex vivo lytic activity comparable to current FDA-approved therapies for leukemia and correlated drug treatment with specific cell–cell contact, cytokine delivery, and leukemia cell lysis. Given the modular nature of these multivalent compounds and their rapid assembly/screening, we anticipate facile extension of this therapeutic approach to a wide range of immune cells, diseased cells, and soluble protein combinations in the future.

中文翻译：

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白血病多价免疫细胞重定向疗法的快速组装和筛选。

与免疫细胞结合并将其细胞毒活性重定向到患病细胞的疗法代表了一种有前景且通用的免疫疗法，可应用于癌症、狼疮和其他疾病；然而，发现这些疗法的传统方法通常是时间密集型的，并且缺乏相关的基于目标的发现方法的吞吐量。受观察到细胞因子 IL-12 可以增强临床批准的 T 细胞重定向疗法 blinatumomab 的抗白血病活性的启发，我们在这里描述了一种嵌合免疫细胞重定向剂的结构和组装，该试剂重定向原代人类的裂解活性T 细胞朝向白血病 B 细胞，同时共同靶向 T 细胞刺激性 IL-12 的递送。我们进一步描述了一种平行组装这些双特异性 T 细胞接合细胞因子 (BiTEokines) 及其高通量表型筛选的组成不同文库的新方法，只需几天的时间即可进行命中识别和组成-功能关系的分析。使用这种方法，我们确定了 CD19 × CD3 × IL12 化合物，其表现出与当前 FDA 批准的白血病疗法相媲美的体外裂解活性，以及​​具有特定细胞 - 细胞接触、细胞因子递送和白血病细胞裂解的相关药物治疗。鉴于这些多价化合物的模块化性质及其快速组装/筛选，我们预计未来这种治疗方法可以轻松扩展到广泛的免疫细胞、患病细胞和可溶性蛋白质组合。