Anti—Gr-1去除新生小鼠髓源性抑制细胞的动态变化  

The dynamic changes of anti-Gr-1 to deplete myeloid-derived suppressor cells in newborn mice

作　　者：林泽锋[1] 童燕陆 王贺珍[1] 付铭[1] 朱德力[1] 张锐忠[1] 夏慧敏[1] 

机构地区：[1]广州医科大学附属广州市妇女儿童医疗中心小儿外科,510120

出　　处：《国际免疫学杂志》2017年第2期117-122,共6页International Journal of Immunology

基　　金：国家自然科学基金（81671498、81600399）

摘　　要：目的分析anti-Gr-1去除新生小鼠髓源性抑制细胞（MDSCs）的动态变化，探讨其去除MDSCs的效率和反应时间。方法将50只BALB／c新生小鼠随机分成5组，每组10只，根据不同时间点腹腔注射anti—Gr-1（15μg）；于出生后第9天收取肝脏、脾脏标本，并将各自充分研磨后的组织悬液通过Percoll液进行梯度离心，分离出其中的单个核细胞；利用流式细胞仪分别检测肝、脾组织的髓源性抑制细胞（CD11b＋Gr-1＋）数量。结果①经anti—Gr-1注射4h后，实验组脾脏中的MDSCs百分比明显低于正常对照组[（9．19±1．40）％比（26．01±2．26）％，t=20．266，P〈0．05]，并能有效维持24～48h，去除率达到64．6％，当48h后MDSCs百分比（16．87±3．49）％，去除率下降至35．1％；②经anti-Gr-1注射24h后，实验组肝脏中的MDSCs百分比与正常对照组比较[（8．98±1．03）％比（15．75±1．10）％，t=11．12，P〈0．05]，具有统计学意义，去除率为42．9％，至48h后MDSCs百分比（4．57±0．95）％，去除率达到70．9％；③同一实验组，脾脏和肝脏各自的M-MDSCs／G—MDSCs比值区别较大。结论Anti—Gr-1对肝脏、脾脏的MDSCs去除效率和反应时间不同：经anti-Gr-1注射后4—48h，脾脏的MDSCs去除率为35．1％～64．6％，而24—48h后肝脏的MDSCs去除率为42．9％-70．9％。Objective To analyze the dynamic changes of myeloid-derived suppressor cells （MDSCs）in newborn mice after the inoculation of anti-Gr-1 antibody and to discuss the efficiency and reaction time of depleting MDSCs. Methods Neonatal BALB/c mice（ n = 50） were randomly separated into 5 groups and were intraperitoneally inoculated with anti-Gr-1 （ 15 μg）. All mice were sacrificed at day 9 after birth, liver and spleen were collected to centrifuge for the mononuclear ceils respectively and MDSCs（ CD11b ＋ Gr-1 ＋） were analyzed through flow cytometry. Results （1）Compared with the normal mice, MDSCs in the spleen of the experimental mice, which were inoculated with anti-Gr-1 for 4 hours, were significantly reduced [ （9.19 ± 1.40）% vs. （26.01 ± 2.26）% ,t = 20. 266, P 〈 0.05 ] and still remained for 24 - 48 hours. The ratio of depleting MDSCs reached at 64.6% but after 48 hours the ratio declined to 35.1%. （2）Compared with the normal mice, MDSCs in liver of the experimental mice which were inoculated with anti-Gr-1 for 24 hours, were significantly reduced[ （8.98 ±1.03） % vs. （15.75 ± 1.10）% ,t = 11.12 ,P 〈0.051. The ratio of depleting MDSCs reached 42.9%. After 48 hours the ratio increased to 70.9%. （3）Compared with the same experimental gourp,the ratio of MDSCs subsets （M-MDSCs/G-MDSCs） in murine liver and spleen were obviously different. Conclusion The efficient and re- acted time of depleting MDSCs with anti-Gr-1 are different in murine liver and spleen. After inoculating with an- ti-Gr-1 ,it will retain the levels of 35.1% - 64.6% MDSCs in murine spleen between 4 and 48 hours, but it will retain the levels of 42.9% - 70.9% MDSCs in murine liver between 24 and 48 hours.

关 键 词：髓源性抑制细胞 固有免疫 Anti-Gr-1 新生小鼠 流式细胞术 

分 类 号：R392[医药卫生—免疫学]