摘要
目的:研究PD-L1在人胎盘源间充质干细胞(Human placenta mesenchymal stem cell,hPMSCs)介导的对脐血CD8+T细胞活化、周期及对IL-17分泌免疫调节中的作用。方法:RT-PCR及FCM检测hPMSCs对PD-L1的表达;应用化学合成的PD-L1 siRNA阻断PD-L1在hPMSCs上的表达;免疫磁珠分选脐血CD8+T细胞;FCM分析阻断PD-L1后,hPMSCs对PHA刺激下CD8+T细胞活化、周期及PMA活化下CD8+T细胞分泌IL-17的影响。结果:hPMSCs高表达PD-L1分子,PD-L1 siRNA能有效阻断hPMSCs对PD-L1的表达;FCM分析结果显示,hPMSCs能够抑制CD8+T细胞对CD69的表达,但阻断PD-L1后,CD69的表达与未阻断组相比无明显变化;与未阻断组相比,处于G0/G1期的CD8+T细胞数量明显减少,处于S期的细胞数量明显增加;在hPMSCs存在条件下,脐血CD8+T细胞对IL-17的分泌明显增加,阻断PD-L1的表达后,IL-17的分泌被进一步上调。结论:PD-L1在hPMSCs上表达能够协同hPMSCs对脐血CD8+T细胞周期的抑制,并且能够抑制hPMSCs上调CD8+T细胞对IL-17的分泌。
Objective：To investigate the effect of PD-L1 in human placenta derived mesenchymal stem cells（hPMSCs） mediating immunoregulation on cord blood CD8＋T cell activation,cell cycle and secretion of IL-17.Methods：The expression of the PD-L1 on hPMSCs was detected by RT-PCR and FCM respectively.Specific PD-L1 siRNAs were transfected into hPMSCs via cathodolyte liposome transfection method.CD8＋T cells were sorted from cord blood with immunomagetic beads.The expression of early activation phenotype,cell cycle and cytokine secretion of cord blood CD8＋T cells were analyzed by FCM.Results：PD-L1 siRNA could effectively block the expression of PD-L1 which was highly expressed on hPMSCs.The expression of CD69 on cord blood CD8＋T cells had no significantly difference between the blocking groups and the unblocking groups.Compared with the unblocking groups,the number of cord blood CD8＋T cells in G0/G1 phase was decreased while the number of cord blood CD8＋T cells in S phase was increased in the blocking groups.hPMSCs caused a sharp increase of IL-17 secretion which was further up-regulated in blocking group.Conclusion：PD-L1 expressed on hPMSCs could promote the inhibitory effect of hPMSCs on cord blood CD8＋T cell cycle,and inhibit the up-regulation of hPMSCs mediated on the expression level of IL-17 secreted by cord blood CD8＋T cells.
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2012年第3期221-225,230,共6页
Chinese Journal of Immunology
基金
山东省科技发展计划资助(2011GGH21818)
山东省自然科学基金资助(Y2006C2)
山东省医药卫生发展计划资助(2007HZ039)
滨州医学院科研启动基金资助(BY2007KYQD09)
