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Characterization of novel dual tandem CD19/BCMA chimeric antigen receptor T cells to potentially treat multiple myeloma

机译:新型双串联CD19 / BCMA嵌合抗原受体T细胞的表征可潜在治疗多发性骨髓瘤

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摘要

Generation and characterization of tan-CAR T cells. ( ) Schematic representation of the plasmid design used to construct single and tandem scFv-CAR-modified T cells. The tan-CAR construct encodes a tandem fusion of the BCMA-scFv and CD19-scFv domains, a CD28 costimulatory domain, and a CD3 zeta-mediated activation signaling domain. ( ) Analysis of transduction efficiency. Isolated and activated human T cells were transduced with the lentiviral vectors CD19-CAR, BCMA-CAR, or tan-CAR. Transduction efficiency was evaluated by flow cytometry using fluorescent-tagged protein L to detect variable light chain Ig together with human CD19 and human BCMA protein. ( ) Quantitative evaluation of CD4 /CD8 T-cell ratios. CAR-positive CD4 T cells and CAR-positive CD8 T cells were detected by flow cytometry. Data are representative of three donors
机译:tan-CAR T细胞的生成和表征。 ()用于构建单个和串联scFv-CAR修饰的T细胞的质粒设计的示意图。 tan-CAR构建体编码BCMA-scFv和CD19-scFv结构域,CD28共刺激结构域和CD3 Zeta介导的激活信号结构域的串联融合。 ()转导效率分析。用慢病毒载体CD19-CAR,BCMA-CAR或tan-CAR转导分离的和活化的人T细胞。使用荧光标记的蛋白L通过流式细胞术评估转导效率,以检测可变轻链Ig以及人CD19和人BCMA蛋白。 ()定量评估CD4 / CD8 T细胞比率。通过流式细胞术检测CAR阳性CD4 T细胞和CAR阳性CD8 T细胞。数据代表了三个捐助者

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