粒细胞集落刺激因子诱导的供者外周血中两种不同亚群树突状细胞免疫生物学特性的研究

目的 从粒细胞集落刺激因子(G-CSF)诱导的供者外周血中分离出CD1c+的髓细胞性DC(MDC)、CD304+类浆细胞样DC(PDC)两类DC亚群,并对其生物学特性进行分析和研究.方法 免疫磁珠法分离出MDC、PDC,分别用TNF-α、CpG2006OND诱导成熟,检测各DC亚群的表型、对同种异基因淋巴细胞的刺激反应.结果 分选的DC纯度均＞95%,MDC表达HLA-DR、CD11c、CD33、CD4,PDC表达HLA-DR、CD4、CD45RA,诱导成熟的两类DC的CD40、CD80表达均明显增高;混合淋巴细胞反应(MLR)显示:MDC具有很强的刺激淋巴细胞增殖能力,PDC刺激能力很弱;各组上清液中IFN-r均增加,MDC、mMDC组增高最为明显;PDC、mPDC刺激组上清液中IL-10明显增高;MDC、mMDC刺激后,胞浆内表达IFN-r的CD4+T细胞均明显增高;PDC、mPDC有上调CIM+CD25high调节性T细胞(Treg)作用;Foxp3 mRNA的表达在各组间无明显的差异.结论 采集的供者外周血中两类DC亚群虽然HLA-DR高表达,但仍处于非成熟状态,在合适的条件下分化成熟;无论MDC成熟与否均表现出很强的刺激T细胞增殖能力,使T细胞分泌IFN-r增加,其分泌的增加可能是促进向TH1极化的结果.PDC可能并不像MDC一样有效地捕获、处理和负载抗原到MHC分子上,因此提呈抗原效率低,表现出对T细胞的弱刺激增殖特性;PDC虽然也可以促进T细胞分泌IFN-r,但似乎并非是通过TH1细胞;PDC并不能促进TH2类因子IL-4的分泌,对TH2的极化作用可能表现在IL-10的分泌上;PDC有诱导CD4+CD25highTreg的作用.

Immunobiologic study of two different isolated DC subsets from the donons' peripheral blood induced by G-CSF

Objective To separate the CD1c+ MDC and CD304 PDC subsets and analyze their immunobiologic properties.Methods MDC or PDC were separated by immunomagnetic beads and then induced by TNF-α or CpG2006 OND.The phenotype of different DC subsets and allogeneic T-cell-stimulating capacity were detected,respectively.Results The purlty of separated DC was about ninety-five percent.MDC expressed HLA-DR,CD11c,CD4 and CD33,PDC expressed HLA-DR,CD4 and CD45RA,and the percentage of CD40,CD80 were up-regulation after induced to mature.The results of mixed lymphocyte reaction(MLR):MDC had strong potential to stimulate allogeneic T lymphocytes proliferation,while PDC had the weak stimslate potential.The supematant IFN-r in all groups was increased.which the groups of MDC and mMDC were more obviously.The supematant IL-10 was increased obviously in groups of PDC and mPDC.The expression of IFN-r in CD4+ cells were increased obviously after stimulated by MDC or mMDC.PDC and mPDC had the effect of inducing of CIM+ CD25high regulatory T lymphocyte in MLR.The expressing of Foxp3 mRNA had no evident difference in all groups.Conclusion Two different DC subsets in the donors peripheral blood which mobilized with G-CSF were in the state of immature and could be induced to mature in the suitable condition.although expressed high constitutive levels of HLA,DR.No matter how MDC was mature or not,they had strong potential to stimulate allogeneic T lymphocytes proliferation.MDC enhanced the production of IFN-r among T lymphocytes,and the increase of IFN-r was probably the result of TH1 polarization.Unlike MDC,PDC was less efficient present antigens and had poor T-cell-stimulating capacity,probably because of their lesser ability to phagocytose,process and present antigens to the MHC molecules.Although PDC could promote the production of IFN-r,it seemed that did not produced by TH 1.PDC could not promote TH2 cells to product IL-4,and the polarization of TH2 probably reflect the production of IL-10.PDC could induce the generation of CD4+ CD25high regulatory T cells.