摘要
目的:研究扩增活化的淋巴细胞在重度免疫缺陷小鼠体内的分布情况。方法:使用96只NPG小鼠,随机分为溶媒对照组和给药组,溶媒对照组动物尾静脉注射给予溶媒1次;给药组动物尾静脉注射给予DiR标记的活化扩增的淋巴细胞(expanded activated lymphocytes,EAL)细胞1次,于给药后不同时间点采血,并使用活体成像的方法检测EAL细胞。在给药后1 h,3 h,2 d,7 d,14 d,28 d,42 d,56 d共8个时间点小鼠实施麻醉后,解剖取血液、心脏、肝脏、脾脏、肺脏、肾脏、脑、睾丸、附睾、子宫、卵巢、胃、十二指肠、结肠、骨髓、脂肪、骨骼肌,冻存。采用流式细胞术、活体成像方法和qPCR方法,研究EAL细胞在外周血以及上述组织器官中的分布情况。结果:给药后2 d外周血中CD_(3)^(+)T细胞、CD_(3)^(+)CD_(4)^(+)T细胞和CD_(3)^(+)CD_(8)^(+)T细胞数量较高,随后呈下降趋势,14 d时最低,其后一直维持较低水平。由动物活体成像结果可知,细胞在小鼠体内主要分布在肺脏、肝脏脾脏部位和双侧腿骨,给药后1 d内主要分布在肺脏和肝脏。由qPCR检测结果可知,细胞主要在肺脏和血液分布最多,其次为脾脏、肝脏和骨髓,其他组织仅有较少分布。给药后1 h,肺脏中细胞基因拷贝数较高,给药后2 d,血液和脾脏中细胞基因拷贝数较高,后都逐渐下降,14 d最低,至56 d时仅有1只动物的个别脏器检测到细胞。结论:EAL细胞尾静脉注射给予NPG小鼠后,主要分布在肺脏、血液和脾脏。给药后56 d时,细胞在动物体内基本消除。
Objective:To study the distribution of expanded activated lymphocytes in severe immunodeficient mice.Methods:Ninety⁃six NPG mice were used and randomly divided into vehicle control group and administration group.The animals in the vehicle control group were given vehicle by tail vein injection once;the animals in the administration group were given DiR⁃labeled EAL cells by tail vein injection once.Blood was collected at different time points after administration,and EAL cells were detected by in vivo imaging system.At 1 h,3 h,2 d,7 d,14 d,28 d,42 d,and 56 d after administration,the mice were anesthetized,and blood,heart,liver,spleen,lung,kidney,brain,testis,epididymis,uterus,ovary,stomach,duodenum,colon,bone marrow,fat,and skeletal muscle were taken for cryopreservation.The distribution of EAL cells in peripheral blood and the above⁃mentioned tissues was studied by flow cytometry,in vivo imaging method and qPCR method.Results:The numbers of CD_(3)^(+)T cells,CD_(3)^(+)CD_(4)^(+)T cells and CD_(3)^(+)CD_(8)^(+) T cells in peripheral blood were higher at 2 d after administration,and then showed a downward trend,reaching the lowest at 14 d,and maintained a low level thereafter.The results of in vivo imaging showed that the cells were mainly distributed in the lungs,liver,spleen and bilateral leg bones,and were mainly distributed in the lungs and liver within 1 d after administration.The results of gene copy number detection showed that the cells were mainly distributed in the lungs and blood,followed by the spleen,liver and bone marrow,and only a little in other tissues.At 1 h after administration,the gene copy number of cells in the lungs was relatively high.At 2 d,the number of gene copies in the blood and spleen was relatively high,and then gradually decreased,reaching the lowest at 14 d,and at 56 d cells were only detected in individual organs of one mice.Conclusion:EAL cells were administered to NPG mice,the cells were mainly distributed in lung,blood and spleen.At 56d after administration,the cells were
作者
侯田田
李雪娇
姜华
秦超
姚志伟
霍艳
耿兴超
黄瑛
HOU Tian-tian;LI Xue-jiao;JIANG Hua;QIN Chao;YAO Zhi-wei;HUO Yan;GENG Xing-chao;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;SanQ(Beijing)Biotechnology Co.,Ltd.,Beijing 100176,China)
出处
《中国新药杂志》
CAS
CSCD
北大核心
2023年第20期2051-2057,共7页
Chinese Journal of New Drugs
基金
国家重点研发计划课题:干细胞产品非临床有效性和安全性评价新技术及规范研究(2021YFA1101602)
中国科学院战略先导科技专项(XDA1604050202)
中国食品药品检定研究院关键技术研究基金资助项目(GJJS⁃2022⁃6⁃1)。
