Department of Transplantation-Immunology, Institute of Immunology, University of Heidelberg, Im Neuenheimer Feld 305, 69120 Heidelberg, Germany.
Transpl Immunol. 2012 Oct;27(2-3):114-21. doi: 10.1016/j.trim.2012.08.003. Epub 2012 Aug 27.
IFNγ-producing CD3(+)CD4(+)CD25(+)Foxp3(+) induced Treg are more frequently detectable in patients with good than in patients with impaired long-term kidney graft function. We investigated the in-vitro function of separated CD3(+)CD4(+)CD25(+)Foxp3(+)IFNγ(+) PBL that were induced by phorbol-12-myristate-13-acetate(PMA)/Ionomycin or alloantigenic stimulation. Additionally, we studied iTreg induction and cell proliferation in MLC with pretransplant obtained PBL. CD4(+)CD25(+)IFNγ(+) PBL separated from PMA/Ionomycin-stimulated PBL of healthy controls inhibited secondary cell cultures of autologous PBL. Furthermore, CD4(+)CD25(+)IFNγ(+) PBL separated from primary MLC and added to secondary MLC suppressed allogeneic T-cell activation in secondary MLC unspecifically, irrespective of the stimulator cell. However, the strongest suppression was observed in specific MLC. Patients with poor long-term graft outcome were able to form IFNγ(+) iTreg in pretransplant MLC. Eight patients with a serum creatinine level ranging from 0.9 to 14 mg/dl 18-29 years posttransplant were studied. In MLC with pretransplant obtained recipient and donor cells, strong IFNγ(+) iTreg (p=0.007) and strong blast induction (p=0.047) were associated with impaired long-term graft outcome. Long-term graft outcome was not associated with cell proliferation and iTreg induction in unspecific MLC with third-party cells as stimulator. The data indicate that patients with impaired long-term graft outcome are able to form high numbers of IFNγ(+) iTreg in specific pretransplant MLC. Quantity of induced IFNγ(+) iTreg depends on the strength of the alloresponse and both parameters are inversely associated with long-term graft outcome.
IFNγ 产生的 CD3(+)CD4(+)CD25(+)Foxp3(+)诱导的 Treg 在长期肾功能良好的患者中比在受损的患者中更频繁地检测到。我们研究了通过佛波醇-12-肉豆蔻酸-13-乙酸酯（PMA）/离子霉素或同种抗原刺激分离的 CD3(+)CD4(+)CD25(+)Foxp3(+)IFNγ(+)PBL 的体外功能。此外，我们研究了在同种异体反应中使用移植前获得的 PBL 进行的 MLC 中的 iTreg 诱导和细胞增殖。从健康对照者的 PMA/Ionomycin 刺激的 PBL 中分离出的 CD4(+)CD25(+)IFNγ(+)PBL 抑制了自身 PBL 的次级细胞培养。此外，从原发性 MLC 中分离出的 CD4(+)CD25(+)IFNγ(+)PBL 并添加到次级 MLC 中，可在次级 MLC 中特异性地抑制同种异体 T 细胞的激活，而与刺激细胞无关。然而，在特异性 MLC 中观察到最强的抑制作用。长期移植物预后不良的患者能够在移植前的 MLC 中形成 IFNγ(+)iTreg。研究了 18-29 年移植后血清肌酐水平在 0.9 至 14mg/dl 之间的 8 名患者。在使用移植前获得的受者和供体细胞的 MLC 中，强烈的 IFNγ(+)iTreg（p=0.007）和强烈的 blast 诱导（p=0.047）与长期移植物预后不良相关。长期移植物预后与使用第三方细胞作为刺激物的非特异性 MLC 中的细胞增殖和 iTreg 诱导无关。这些数据表明，长期移植物预后不良的患者能够在特定的移植前 MLC 中形成大量的 IFNγ(+)iTreg。诱导的 IFNγ(+)iTreg 的数量取决于同种异体反应的强度，这两个参数与长期移植物预后呈负相关。
