嵌合抗原受体T细胞在重度免疫缺陷鼠体内的生物分布研究  

Biodistribution study of CAR-T cells in non-bearing-tumor and tumor-bearing severe immunodeficient mice

作　　者：侯田田 李雪姣 孙磊 秦超 赵晶 霍艳[1] 王歈 耿兴超[1] 黄瑛[1] HOU Tiantian;LI Xuejiao;SUN Lei;QIN Chao;ZHAO Jing;HUO Yan;WANG Yu;GENG Xingchao;HUANG Ying(National Center for Safety Evaluation of Drugs,National Institutes for Food and Drug Control,Beijing Key Laboratory for Safety Evaluation of Drugs,Beijing 100176,China;Immunotech Applied Science Limited,Beijing 100176,China)

机构地区：[1]中国食品药品检定研究院国家药物安全评价监测中心,药物非临床安全评价研究北京市重点实验室,北京100176 [2]北京永泰生物制品有限公司,北京100176

出　　处：《中国药物警戒》2022年第8期823-827,844,共6页Chinese Journal of Pharmacovigilance

基　　金：国家重点研发计划(2021YFA1101602);中国科学院战略先导科技专项(XDA1604050202)。

摘　　要：目的研究嵌合抗原受体T(CAR-T)细胞在非荷瘤和荷瘤重度免疫缺陷小鼠体内的增殖、分布和存续时间。方法非荷瘤小鼠分布研究:使用转染荧光素酶的CAR-T(CAR-T-FLuc)细胞于小鼠尾静脉给药1次,于给药后不同时间点采用活体成像的方法检测CAR-T-FLuc细胞分布情况,并在给药后3 h和2、7、14、28、56、84 d解剖动物,取血液、心脏、肝脏、脾脏、肺脏、肾脏、脑、睾丸、附睾、子宫、卵巢、胃、十二指肠、骨髓、脂肪、骨骼肌,QPCR方法检测CAR-T-FLuc细胞在外周血及上述组织器官中的分布情况。荷瘤小鼠分布研究:小鼠尾静脉注射Raji细胞,建立肿瘤模型,之后给予CAR-T-FLuc细胞,并采用与非荷瘤小鼠相同的检测方法考察CAR-T-FLuc细胞在活体和各组织脏器不同时间点的表达。结果非荷瘤小鼠CAR-T-FLuc细胞在脾脏和肺脏中分布最高,其次是血液;给药后3 h,细胞主要分布在肺脏,后转移到其他脏器;各脏器给药后2、14、56 d是CAR表达较高时间点,84 d时CAR表达接近为零。荷瘤小鼠CAR-T-FLuc细胞在脾脏中分布最高,其次是肺和血液。给药后5 min外周血中即可检测到CAR表达,1、6、27 d为CAR表达高峰时间点;大多组织于给药后3 h可检测到CAR-T细胞,以肺中含量最高,随后呈下降趋势,15 d时最低,之后各组织CAR-T细胞含量持续升高,55 d达高峰。结论CAR-T-FLuc细胞在非荷瘤鼠和荷瘤鼠中主要分布在脾脏、肺脏和血液,但在2种动物体内分布具有不同变化趋势,且在荷瘤鼠体内扩增水平高于非荷瘤鼠。Objective To investigate the proliferation,distribution and persistance of chimeric antigen receptor T(CAR-T)cells in non-bearing-tumor and tumor-bearing severe immunodeficient mice.Methods Non-bearing-tumor mice:The mice received intravenous infusions of CAR-T-FLuc cells that were transduced with firefly luciferase.CAR-T-FLuc cells were monitored by in vivo imaging system at different time points.The animals were sacrificed at 3 hours,2 days,7 days,14 days,28 days,56 days,84 days respectively.Blood,heart,liver,spleen,lung,kidney,brain,testis,epididymis,uterus,ovary,stomach,duodenum,bone marrow,fat,skeletal muscle were taken successively.By QPCR,the distribution of CAR-T-FLuc cells were detected in these apparatuses and peripheral blood.Tumor-bearing mice:A tumor model was established,mice were injected with Raji cells via the tail vein,and then the mice were intravenous injected with CAR-T-FLuc.The expression of CAR-T-FLuc cells in vivo and at different time points in various tissues were investigated by the same detection method as in non-tumor-bearing mice.Results In non-bearing-tumor mice:CAR-T-FLuc cells were mainly distributed in the lung and spleen,followed by blood.At 3 hours after administration,the cells were mainly distributed in the lung and then transferred to other organs.The expression of CAR of each organ was higher at 2 days,14 days and 56 days.And CAR expression was close to zero at 84 days.In tumor-bearing mice:CAR-T-FLuc cells were mainly distributed in the spleen,followed by the lung and blood.CAR expression was detected in peripheral blood 5 minutes after administration,and 1 day,6 days and 27 days were the peak time of CAR expression.In most tissues,CAR-T cells were detected 3 hours after drug administration,with the highest content in the lungs,followed by a downward trend,and the lowest at 15 days.After that,the CAR-T cell content of each tissue continued to rise,reaching a peak at 55 days.Conclusion CAR-T-FLuc cells are mainly distributed in the spleen,lung and blood in non-tumorbearing

关 键 词：嵌合抗原受体T细胞(CAR-T细胞) 生物分布 NSG小鼠 荷瘤鼠 非荷瘤鼠 重度免疫缺陷 

分 类 号：R979.1[医药卫生—药品]