Function of Novel Anti-CD19 Chimeric Antigen Receptors with Human Variable Regions Is Affected by Hinge and Transmembrane Domains
Function of Novel Anti-CD19 Chimeric Antigen Receptors with Human Variable Regions Is Affected by Hinge and Transmembrane Domains
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DOI：
10.1016/j.ymthe.2017.07.013
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发表时间：
2017-11-01
影响因子：
12.4
通讯作者：
Kochenderfer, James N.
中科院分区：
文献类型：
--
作者：
Alabanza, Leah;Pegues, Melissa;Kochenderfer, James N.
抗CD19嵌合抗原受体（CAR）T细胞已使B细胞恶性肿瘤缓解，但包括细胞因子介导的毒性以及CAR T细胞在体内持续时间短等问题可能会限制抗CD19 CAR - T细胞的有效性。临床上已测试的抗CD19 CARs含有源自鼠抗体的单链可变片段（scFvs）。我们设计并构建了新型抗CD19 CARs，其包含具有完全人源可变区的scFv。表达这些CARs的T细胞能特异性识别CD19⁺靶细胞，并执行包括脱颗粒、细胞因子释放和增殖等功能。我们比较了具有CD28共刺激结构域以及来自人CD28分子或人CD8α分子的铰链和跨膜结构域的CARs。与表达具有CD28铰链和跨膜结构域的CARs的T细胞相比，表达具有CD8α铰链和跨膜结构域的CARs的T细胞产生较低水平的细胞因子，并表现出较低水平的激活诱导的细胞死亡（AICD）。重要的是，具有来自CD8α或CD28的铰链和跨膜区域的CARs在消除小鼠体内已形成的肿瘤方面具有相似的能力。在具有CD28共刺激结构域的抗CD19 CARs中，较低水平的炎性细胞因子产生和AICD是CD8α铰链和跨膜结构域相较于CD28铰链和跨膜结构域潜在的临床优势。
Anti-CD19 chimeric antigen receptor (CAR) T cells have caused remissions of B cell malignancies, but problems including cytokine-mediated toxicity and short persistence of CAR T cells in vivo might limit the effectiveness of anti-CD19 CART cells. Anti-CD19 CARs that have been tested clinically had single-chain variable fragments (scFvs) derived from murine antibodies. We have designed and constructed novel anti-CD19 CARs containing a scFv with fully human variable regions. T cells expressing these CARs specifically recognized CD19(+) target cells and carried out functions including degranulation, cytokine release, and proliferation. We compared CARs with CD28 costimulatory moieties along with hinge and transmembrane domains from either the human CD28 molecule or the human CD8 alpha molecule. Compared with T cells expressing CARs with CD28 hinge and transmembrane domains, T cells expressing CARs with CD8 alpha hinge and trans membrane domains produced lower levels of cytokines and exhibited lower levels of activation-induced cell death (AICD). Importantly, CARs with hinge and transmembrane regions from either CD8 alpha or CD28 had similar abilities to eliminate established tumors in mice. In anti-CD19 CARs with CD28 costimulatory moieties, lower levels of inflammatory cytokine production and AICD are potential clinical advantages of CD8 alpha hinge and transmembrane domains over CD28 hinge and transmembrane domains.