Anti-Gr-1去除新生小鼠髓源性抑制细胞的动态变化

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(15 μg);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24～48 h,去除率达到64.6％,当48 h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48 h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4～48 h,脾脏的MDSCs去除率为35.1％ ～64.6％,而24 ～48 h后肝脏的MDSCs去除率为42.9％～70.9％.

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

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 cells respectively and MDSCs(CD11b + Gr-1 +) were analyzed through flow cytometry.Results ①Compared with the normal mice,MDSCs in the spleen of the experimental mice,which were inoculated with anti-Gr-1 for 4 hours,were significandy 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％.②Compared 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 ＜ 0.05].The ratio of depleting MDSCs reached 42.9％.After 48 hours the ratio increased to 70.9％.③Compared 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 reacted time of depleting MDSCs with anti-Gr-1 are different in murine liver and spleen.After inoculating with anti-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.