Chimeric antigen receptors (CARs) are used to redirect effector cell specificity to selected cell surface antigens. Using CARs, antitumor activity can be initiated in patients with no prior tumor specific immunity. Although CARs have shown promising clinical results, the technology remains limited by the availability of specific cognate cell target antigens. To increase the repertoire of targetable tumor cell antigens we utilized the immune system of the sea lamprey to generate directed variable lymphocyte receptors (VLRs). VLRs serve as membrane bound and soluble immune effectors analogous but not homologous to immunoglobulins. They have a fundamentally different structure than immunoglobulin (Ig)-based antibodies while still demonstrating high degrees of specificity and affinity. To test the functionality of VLRs as the antigen recognition domain of CARs, two VLR-CARs were created. One contained a VLR specific for a murine B cell leukemia and the other contained a VLR specific for the human T cell surface antigen, CD5. The CAR design consisted of the VLR sequence, myc-epitope tag, CD28 transmembrane domain, and intracellular CD3ζ signaling domain. We demonstrate proof of concept, including gene transfer, biosynthesis, cell surface localization, and effector cell activation for multiple VLR-CAR designs. Therefore, VLRs provide an alternative means of CAR-based cancer recognition.

中文翻译：

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使用七lamp鳗来源的可变淋巴细胞受体进行嵌合抗原受体的基因工程。

嵌合抗原受体（CARs）用于将效应细胞特异性重定向至选定的细胞表面抗原。使用CAR，可以在没有先前肿瘤特异性免疫的患者中启动抗肿瘤活性。尽管CARs已显示出令人鼓舞的临床结果，但该技术仍然受到特定同源细胞靶抗原可用性的限制。为了增加可靶向肿瘤细胞抗原的库，我们利用海七lamp鳗的免疫系统来产生定向可变淋巴细胞受体（VLR）。VLR类似于但不与免疫球蛋白同源，可作为膜结合和可溶性免疫效应子。它们具有与基于免疫球蛋白（Ig）的抗体根本不同的结构，同时仍显示出高度的特异性和亲和力。为了测试VLR作为CAR的抗原识别域的功能，创建了两个VLR-CAR。一个包含对鼠B细胞白血病特异的VLR，另一个包含对人T细胞表面抗原CD5特异的VLR。CAR设计由VLR序列，myc-表位标签，CD28跨膜结构域和细胞内CD3ζ信号结构域组成。我们演示了概念验证，包括用于多个VLR-CAR设计的基因转移，生物合成，细胞表面定位和效应细胞激活。因此，VLR提供了一种基于CAR的癌症识别方法。CD28跨膜结构域和细胞内CD3ζ信号结构域。我们演示了概念验证，包括用于多个VLR-CAR设计的基因转移，生物合成，细胞表面定位和效应细胞激活。因此，VLR提供了一种基于CAR的癌症识别的替代方法。CD28跨膜结构域和细胞内CD3ζ信号结构域。我们演示了概念验证，包括用于多个VLR-CAR设计的基因转移，生物合成，细胞表面定位和效应细胞激活。因此，VLR提供了一种基于CAR的癌症识别的替代方法。