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FLAG 标记的 CD19 特异性 CAR-T 细胞消除了携带 CD19 的实体瘤细胞。

FLAG-tagged CD19-specific CAR-T cells eliminate CD19-bearing solid tumor cells and .

机构信息

ProMab Biotechnologies, Richmond, CA.

Department of Infectious Diseases, University of California, Berkeley, CA.

出版信息

Front Biosci (Landmark Ed). 2017 Jun 1;22(10):1644-1654. doi: 10.2741/4563.

Abstract

Autologous T cells expressing chimeric antigen receptors (CARs) specific for CD19 have demonstrated remarkable efficacy as therapeutics for B cell malignancies. In the present study, we generated FLAG-tagged CD19-specific CAR-T cells (CD19-FLAG) and compared them to their non-tagged counterparts for their effects on solid and hematological cancer cells and . For solid tumors, we used HeLa cervical carcinoma cells engineered to overexpress CD19 (HeLa-CD19), and for hematological cancer we used Raji Burkitt's lymphoma cells, which endogenously express CD19. Like non-tagged CD19 CAR-T cells, CD19-FLAG CAR-T cells expanded in culture >100-fold and exhibited potent cytolytic activity against both HeLa-CD19 and Raji cells . CD19-FLAG CAR-T cells also secreted significantly more IFN-gamma and IL-2 than the control T cells. , CD19-FLAG CAR-T cells significantly blocked the growth of HeLa-CD19 solid tumors, increased tumor cleaved caspase-3 levels, and expanded systemically. CD19-FLAG CAR-T cells also significantly reduced Raji tumor burden and extended mouse survival. These results demonstrate the strong efficacy of FLAG-tagged CD19 CAR-T cells in solid and hematological cancer models.

摘要

表达嵌合抗原受体(CAR)的自体 T 细胞特异性针对 CD19,已被证明是治疗 B 细胞恶性肿瘤的有效方法。在本研究中,我们生成了 FLAG 标记的 CD19 特异性 CAR-T 细胞(CD19-FLAG),并将其与非标记的 CAR-T 细胞进行比较,以评估它们对实体瘤和血液系统恶性肿瘤细胞的作用。对于实体瘤,我们使用过表达 CD19 的 HeLa 宫颈癌细胞(HeLa-CD19),对于血液系统恶性肿瘤,我们使用 Raji 伯基特淋巴瘤细胞,其内源表达 CD19。与非标记的 CD19 CAR-T 细胞一样,CD19-FLAG CAR-T 细胞在培养中扩增超过 100 倍,并对 HeLa-CD19 和 Raji 细胞表现出强大的细胞毒性作用。CD19-FLAG CAR-T 细胞还分泌了明显更多的 IFN-γ和 IL-2。此外,CD19-FLAG CAR-T 细胞显著抑制了 HeLa-CD19 实体瘤的生长,增加了肿瘤裂解的 caspase-3 水平,并在体内扩增。CD19-FLAG CAR-T 细胞还显著减少了 Raji 肿瘤负担并延长了小鼠的存活时间。这些结果表明,FLAG 标记的 CD19 CAR-T 细胞在实体瘤和血液系统恶性肿瘤模型中具有强大的疗效。

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