中国药物警戒 ›› 2022, Vol. 19 ›› Issue (8): 823-827.
DOI: 10.19803/j.1672-8629.2022.08.03

• CAR-T 细胞非临床安全性研究与评价专栏 • 上一篇    下一篇

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

侯田田1, 李雪姣2, 孙磊2, 秦超1, 赵晶1, 霍艳1, 王歈2, 耿兴超1#, 黄瑛1*   

  1. 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
  • 收稿日期:2022-04-29 出版日期:2022-08-15 发布日期:2022-08-15
  • 通讯作者: *黄瑛,女,博士,副研究员,药物临床前安全性评价。E-mail：huangying1002@nifdc.org.cn;#为共同通信作者。
  • 作者简介:侯田田,女,硕士,助理研究员,药物临床前安全性评价。
  • 基金资助:
    国家重点研发计划（2021YFA1101602）; 中国科学院战略先导科技专项（XDA1604050202）

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

HOU Tiantian1, LI Xuejiao2, SUN Lei2, QIN Chao1, ZHAO Jing1, HUO Yan1, WANG Yu2, GENG Xingchao1#, HUANG Ying1*   

  1. 1National Center for Safety Evaluation of Drugs, National Institutes for Food and Drug Control, Beijing Key Laboratory for Safety Evaluation of Drugs, Beijing 100176, China;
    2Immunotech Applied Science Limited, Beijing 100176, China
  • Received:2022-04-29 Online:2022-08-15 Published:2022-08-15

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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。

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

Abstract: 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-tumor-bearing mice and tumor-bearing mice, but the distribution trends in both animals are different. The amplification level in tumor-bearing mice was higher than that in non-tumor-bearing mice.

Key words: chimeric antigen receptor T(CAR-T) cells, biodistribution, NSG mice, tumor-bearing mice, non-bearing-tumor mice, severe immunodeficiency

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