摘要目的 探讨人羊膜间充质细胞(human amniotic mesenchymal cells,hAMCs)对异体外周血淋巴细胞的免疫抑制作用,阐明人羊膜间充质细胞调节移植免疫排斥的作用机制.方法 从新鲜足月剖宫产废弃的羊膜组织分离出hAMCs并进行鉴定.采用密度梯度离心从健康志愿者外周血中分离出单个核淋巴细胞(PBMLs),在植物血凝素(PHA)刺激下,分别按1∶0.05,1∶0.10,1∶0.20的比例以hAMCs处理,MTS法检测淋巴细胞的增殖,ELISA法检测细胞因子IFN-γ、TNF-α的分泌.结果 hAMCs能明显抑制异体外周血淋巴细胞的增殖,hAMCs与PBMLs比值为0.05∶1,0.10∶1,0.20∶1时的增殖抑制率分别为16.91％、20.83％、28.19％.hAMCs还可显著抑制异体淋巴细胞IFN-γ和TNF-α的分泌(P＜0.01),并且在一定范围内随着hAMCs数量的增加抑制作用更加明显.结论 hAMCs能抑制异体淋巴细胞增殖并降低淋巴细胞IFN-γ和TNF-α的分泌,这可能是其预防和减轻移植排斥反应发生的机制之一.

abstractsObjective To explore the immunoregulatory effects of human amniotic mesenchymal cells (hAMCs) on allogeneic peripheral blood lymphocytes.Methods The hAMCs were isolated from abandoned human amnion. Peripheral blood mononuclear lymphocytes (PBMLs) were separated from healthy donors by density gradient centrifugation. Then,PBMLs were treated with phytohemagglutinin (PHA) and different concentrations of hAMCs.Proliferation effect of PBMLs was tested using MTS assay,and production of IFN-γ and TNF-α by PBMLs was detected by ELISA. Results hAMCs could remarkably inhibit the lymphocytes proliferation.When the ratios of hAMCs to PBMLs were 0.05∶ 1,0.10∶ 1,0.20∶ 1,.the inhibitory rates of PBMLs proliferation were 16.91％,20.83％ and 28.19％,respectively.HAMCs also decreased the production of IFN-γ and TNF-α by PBMLs in a dose-dependent manner (P ＜0.01).Conclusions HAMCs could inhibit the proliferation of allogeneic lymphocytes and reduce secretion of IFN-γ and TNF-α,which might be one of the mechanism for prevention and remission of transplant rejection.