雷帕霉素对CAR-T细胞功能的影响

Effects of Rapamycin on the Function of CAR-T cells

为探索雷帕霉素对CAR-T细胞体外杀伤靶细胞、杀伤后的扩增及细胞因子分泌能力的影响,该研究在体外使用靶向CD19的CAR-T细胞与CD19^(+)CD22^(+)Raji靶细胞的T细胞特异性杀伤模型,在不同效靶比、不同时相点、不同雷帕霉素浓度下,通过流式细胞绝对计数检测细胞杀伤率发现雷帕霉素不影响CAR-T细胞对靶细胞的杀伤能力。通过羟基荧光素二醋酸盐琥珀酰亚胺脂(CFSE)检测CAR-T细胞扩增能力,发现雷帕霉素通过抑制CAR-T细胞增殖而非促进凋亡的方式降低其活化后的扩增数量。通过流式细胞术微球阵列法(CBA)检测细胞因子分泌情况,发现雷帕霉素可显著性降低CAR-T细胞被靶细胞活化后的细胞因子分泌水平。综上所述,雷帕霉素可在不影响CAR-T细胞杀伤功能的前提下,降低其炎性因子分泌水平。

To investigate the effects of rapamycin on the specific killing of CAR-T(chimeric antigen recep­tor T)cells,the expansion and cytokine secretion of CAR-T cells in vitro.This study used a T cell specific killing model in which the effect cells were the CAR-T targeting CD19 and the target cells were CD19^(+)CD22^(+)Raji cells.Under different effect-target ratio with various rapamycin concentration at various drug administration time,the specific killing rates of CAR-T were detected by flow cytometry absolute count and it was found that rapamycin did not affect the killing ability of CAR-T cells.Using CFSE(carboxyfluorescein diacetate succinimidyl ester)to detect the expansion ability of CAR-T cells,it was found that rapamycin could slow down the increase rate of CAR-T cells number by inhibiting their proliferation rather than promoting apoptosis.Using flow cytometry CBA(cytometric bead array)to detect cytokine secretion,it was found that rapamycin could significantly inhibited cytokines secre­tion level of CAR-T cells after activation by target cells.In summary,rapamycin can reduce the secretion level of inflammatory cytokines without affecting the killing function of CAR-T cells.