CAJ | 학술논문

目的研究程序性死亡配体2(programmed death ligand 2,PDL2)在人胎盘源间充质干细胞(human placenta mesenchymal stem cell,hPMSCs)对外周血T细胞活化、增殖及周期免疫调节中的作用.方法 RT-PCR、激光共聚焦(laser scanning confocal microscopy,LSCM)及流式细胞术(flow cytometry,FCM)检测PDL2在hPMSCs上的表达；应用PDL2 siRNA阻断PDL2在hPMSCs上的表达；密度梯度离心法分离纯化T细胞；FCM分析阻断PDL2后,hPMSCs对植物血凝素(PHA)刺激下T细胞活化及波佛脂(PMA)活化下T细胞增殖和周期的影响.结果 hPMSCs高表达PDL2分子,PDL2siRNA能有效阻断PDL2在hPMSCs上的表达；FCM分析结果显示,hPMSCs能够抑制CD69在T细胞上的表达,但阻断PDL2后,CD69的表达与未阻断组相比无明显变化；hPMSCs能够显著抑制PMA活化的T细胞的增殖,且阻断PDL2后,其增殖指数被进一步上调；与未阻断组相比,处于G0/G1期的T细胞数量明显减少,处于S期的细胞数量明显增加.结论 PDL2在hPMSCs上表达能够协同hPMSCs对外周血T细胞周期的抑制,进而抑制T细胞的增殖.
목적 연구정서성사망배체2(programmed death ligand 2,PDL2)재인태반원간충질간세포(human placenta mesenchymal stem cell,hPMSCs)대외주혈T세포활화、증식급주기면역조절중적작용.방법 RT-PCR、격광공취초(laser scanning confocal microscopy,LSCM)급류식세포술(flow cytometry,FCM)검측PDL2재hPMSCs상적표체；응용PDL2 siRNA조단PDL2재hPMSCs상적표체；밀도제도리심법분리순화T세포；FCM분석조단PDL2후,hPMSCs대식물혈응소(PHA)자격하T세포활화급파불지(PMA)활화하T세포증식화주기적영향.결과 hPMSCs고표체PDL2분자,PDL2siRNA능유효조단PDL2재hPMSCs상적표체；FCM분석결과현시,hPMSCs능구억제CD69재T세포상적표체,단조단PDL2후,CD69적표체여미조단조상비무명현변화；hPMSCs능구현저억제PMA활화적T세포적증식,차조단PDL2후,기증식지수피진일보상조；여미조단조상비,처우G0/G1기적T세포수량명현감소,처우S기적세포수량명현증가.결론 PDL2재hPMSCs상표체능구협동hPMSCs대외주혈T세포주기적억제,진이억제T세포적증식.
Objective To investigate the effect of programmed death ligand 2 (PDL2) in human placenta derived mesenchymal stem cells(hPMSCs) mediated immunoregulation on peripheral blood T cells activation,proliferation and cell cycle.Methods The expression of the PDL2 on hPMSCs was detected by RT-PCR,LSCM and FCM,respectively.Specific PDL2 siRNAs were transfected into hPMSCs via cathodolyte liposome transfection method.T cells were sorted from healthy peripheral blood by gradient centrifugation.The expression of early activation phenotype,proliferation and cell cycle of T cells were analyzed by FCM.Results PDL2 siRNA could effectively block the expression of PDL2 which was highly expressed on hPMSCs.The expression of CD69 on T cells had no significantly difference in blocking groups compared with unblocking groups.hPMSCs could inhibit the proliferation of T cells induced by PMA,compared with that of unblocking groups,the number of the T cells in G0/G1 phase was decreased while the number of the T cells in S phase was increased in the blocking groups.Conclusion PDL2 expressed on hPMSCs could promote the inhibitory effect of hPMSCs on T cell cycle and proliferation.