人羊膜间充质干细胞与骨髓间充质干细胞对外周血淋巴细胞的免疫调节作用比较

Immunomodulatory effects of human amniotic versus bone marrow-derived mesenchymal stem cells on peripheral blood T lymphocytes in vitro

目的比较人羊膜间充质干细胞(h AMSC)与骨髓间充质干细胞(h BMSC)对异体外周血淋巴细胞的免疫调节功能,为临床应用h AMSC治疗移植物抗宿主病提供实验依据。方法通过酶消化法和Ficoll-Hypaque密度梯度离心法分别分离出人羊膜间充质干细胞和骨髓间充质干细胞,体外扩增培养间充质干细胞(MSC),获取第4代细胞,建立MSC与经植物血凝素(PHA)刺激的异体人外周血单个核细胞(PBMC)共培养体系,采用流式细胞术分析比较两种不同共培养体系中T淋巴细胞亚群的变化情况,采用ELISA法比较两种MSC共培养体系中细胞因子白介素2(IL-2)及白介素10(IL-10)的分泌水平。结果 (1)h AMSC+PBMC+PHA、h BMSC+PBMC+PHA共培养组Treg、Th2、Tc2细胞亚群的比例均较PBMC+PHA组明显上升(P<0.05),Th1、Tc1细胞亚群的比例均较PBMC+PHA组明显下降(P<0.05),且h AMSC+PBMC+PHA、h BMSC+PBMC+PHA共培养组T细胞亚群的变化差异无统计学意义(P>0.05);(2)ELISA结果提示,与PBMC+PHA组相比,h AMSC+PBMC+PHA、h BMSC+PHA+PBMC共培养组上清液中IL-2的水平均明显下降(P<0.05),h AMSC+PBMC+PHA、h BMSC+PHA+PBMC共培养组上清液中IL-10水平均明显上升(P<0.05),h AMSC+PBMC+PHA、h BMSC+PBMC+PHA共培养组上清液中IL-2、IL-10含量的变化无统计学差异(P>0.05)。结论 h AMSC、h BMSC对外周血淋巴细胞具有相似的免疫调节功能,提示h AMSC可能为移植物抗宿主病的预防和治疗提供一种新的选择。

Objective To compare the immunomodulatory effects of human amniotic mesenchymal stem cell （hAMSCs） and human bone marrow mesenchymal stem cells （hBMSCs） on peripheral blood T lymphocytes in an in vitro co-culture system. Methods hAMSCs and hBMSCs isolated using enzymatic digestion and Ficoll-Hypaque density gradient centrifugation, respectively, were culture-expanded in vitro to obtain the 4th-generation cells. The two MSCs were co-cultured separately with human peripheral blood mononuclear cells stimulated with phytohemagglutinin （PHA-PBMSC） to investigate the changes in T lymphocyte subsets using flow cytomety and the production of interleukin-2 （IL-2） and IL-10 by the T lymphocytes using enzyme-linked immunosorbent assay （ELISA）. Results Co-culture with either hAMSCs or hBMSCs significantly increased the proportions of Treg, Th2 and Tc2 and decreased Th1 and Tc1 cell subsets in the PBMCs as compared with the PBMCs cultured alone （P〈0.05）, and the changes in the PBMCs were similar between the two co-culture systems （P〉0.05）. In both of the two co-culture systems, IL-2 production by the lymphocytes was significantly lowered （P〈0.05） and IL-10 production was significantly increased （P〈0.05） as compared with their levels in the PBMCs cultured alone;no significant difference was found in IL-2 or IL-10 levels between the two co-culture systems （P〉0.05）. Conclusion The MSCs derived from human amnion and bone marrow have similar immunomodulatory effects on the T lymphocytes, suggesting the possibility of using hAMSCs in the treatment of graft-versus-host disease after allogeneic hematopoietic stem cell transplantation.