文章：

基于CD30 mAb的CAR修饰T细胞疗法

CAR-modified T-cell therapy based on CD30 mAb 

原文发布日期：2021-01-29 

英文摘要：

Chimeric antigen receptor T-cell immunotherapy (CAR-T) has shown remarkable efficacy in treating tumors of lymphopoietic origin. Herein, we demonstrate an effective CAR-T cell treatment for recurrent and malignant CD30-positive peripheral T-cell lymphomas (PTCL) has been demonstrated. The extracellular fragment gene sequences of CD30 were obtained from tumor tissues of PTCL patients and cloned into a plasmid vector to express the CD30 antigen. The CD30 targeting single-chain antibody fragment (scFv) was obtained from CD30-positive monoclonal hybridoma cells, which were obtained from CD30 antigen immunized mice. After a second-generation of CAR lentiviral construction, CD30 CAR T cells were produced and used to determine the cytotoxicity of this construct toward Karpas 299 cells. The results of CD30 CAR T-mediated cell lysis show that 9C11-2 CAR T cells could significantly promote the lysis of CD30-positive Karpas 299 cells in both LDH and real-time cell electronic sensing (RTCA) assays. In vivo data show that 9C11-2 CAR T cells effectively suppress the tumor growth in a Karpas 299 cell xenograft NCG mouse model. The CD30 CAR T cells exhibited an efficient cytotoxic effect after being co-cultured with the target cells and they also exhibited a significant tumor-inhibiting ability after being intravenously injected into PTCL xenograft tumors; these observations suggest that the new CD30 CAR-T cell may be a promising therapeutic candidate for cancer therapy. 

摘要翻译：

嵌合抗原受体T细胞免疫疗法（CAR-T）在治疗淋巴造血来源肿瘤方面已展现出显著疗效。本研究证实了一种针对复发性恶性CD30阳性外周T细胞淋巴瘤（PTCL）的有效CAR-T细胞疗法。我们从PTCL患者肿瘤组织中获取CD30胞外片段基因序列，将其克隆至质粒载体以表达CD30抗原；同时通过CD30抗原免疫小鼠获得单克隆杂交瘤细胞，从中分离出靶向CD30的单链抗体片段（scFv）。成功构建第二代CAR慢病毒载体后，我们制备了CD30 CAR-T细胞，并评估其对Karpas 299细胞的细胞毒性作用。LDH释放实验和实时细胞电子传感系统（RTCA）检测结果显示，9C11-2 CAR-T细胞能显著促进CD30阳性Karpas 299细胞的裂解。体内实验表明，9C11-2 CAR-T细胞在Karpas 299细胞异种移植NCG小鼠模型中有效抑制了肿瘤生长。该CD30 CAR-T细胞与靶细胞共培养后表现出高效细胞毒性，静脉注射至PTCL异种移植瘤模型后亦显示出显著抑瘤能力，表明新型CD30 CAR-T细胞有望成为癌症治疗的候选方案。

原文链接：

CAR-modified T-cell therapy based on CD30 mAb