Department of Internal Medicine, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands.
Transplantation. 2010 Mar 27;89(6):655-66. doi: 10.1097/TP.0b013e3181c9cc7a.
BACKGROUND.: Rabbit antithymocyte globulins (rATGs) are known to convert CD4CD25FoxP3 T cells from healthy individuals to CD4CD25FoxP3 T cells. In this study, we investigated the effect of rATG on the induction of regulatory T cells (Tregs) from blood cells of patients with end-stage renal disease who are candidates for transplantation and rATG-induction therapy. The induced Tregs were analyzed and compared with naturally occurring CD4CD25FoxP3T cells. METHODS.: The CD25 T cells of pretransplant patients (n=7) and healthy controls (n=4) were stimulated with rATG or control rabbit immunoglobulins for 24 hr. The phenotype of induced Tregs was examined by flow cytometry, and their function was studied in the conventional suppression assay. Further characterization was performed by mRNA analyses. RESULTS.: After 24 hr, the percentage of CD4CD25FoxP3CD127 T cells and CD8CD25FoxP3CD127 T cells became higher in the rATG-treated samples compared with the rabbit immunoglobulin-treated samples (P<0.01). The rATG-induced CD25T cells, whether CD4 or CD8 inhibited the allogeneic responses of CD25 effector T cells as vigorously as natural CD25T cells. However, the proportion of FoxP3 within the top 2% rATG-induced CD4CD25T-cells was lower than within the natural CD4CD25T-cells (11%+/-2% vs. 95%+/-5%, P<0.01). The mRNA-expression levels of interleukin-27, interleukin-10, interferon-gamma, perforin, and granzyme B were markedly higher compared with natural CD25T-cells (all P=0.03), whereas CTLA4 (P=0.03), transforming growth factor-beta (P=0.02), and RORgammat (P=0.04) were lower. CONCLUSION.: rATG allows the induction of Tregs from patient peripheral blood mononuclear cell in vitro. In comparison with natural Tregs, the rATG-induced Tregs are phenotypically distinct but have similar regulatory activities. rATG may beneficially contribute to the mechanisms that control alloreactivity.
兔抗胸腺细胞球蛋白（rATG）已知可将健康个体的 CD4CD25FoxP3 T 细胞转化为 CD4CD25FoxP3 T 细胞。在这项研究中，我们研究了 rATG 对接受移植和 rATG 诱导治疗的终末期肾病患者血细胞中诱导调节性 T 细胞（Tregs）的影响。分析了诱导的 Tregs，并与天然存在的 CD4CD25FoxP3T 细胞进行了比较。方法：在移植前患者（n=7）和健康对照者（n=4）的 CD25 T 细胞中，用 rATG 或对照兔免疫球蛋白刺激 24 小时。通过流式细胞术检测诱导 Tregs 的表型，并在常规抑制试验中研究其功能。通过 mRNA 分析进行进一步表征。结果：24 小时后，与兔免疫球蛋白处理的样本相比，rATG 处理的样本中 CD4CD25FoxP3CD127 T 细胞和 CD8CD25FoxP3CD127 T 细胞的比例更高（P<0.01）。rATG 诱导的 CD25T 细胞，无论是 CD4 还是 CD8，都像天然 CD25T 细胞一样强烈抑制同种异体反应性 CD25 效应 T 细胞。然而，rATG 诱导的 CD4CD25T 细胞中 FoxP3 在前 2%的比例低于天然 CD4CD25T 细胞（11%+/-2%比 95%+/-5%，P<0.01）。与天然 CD25T 细胞相比，白细胞介素-27、白细胞介素-10、干扰素-γ、穿孔素和颗粒酶 B 的 mRNA 表达水平明显升高（均 P=0.03），而 CTLA4（P=0.03）、转化生长因子-β（P=0.02）和 RORgammat（P=0.04）则较低。结论：rATG 可在体外诱导患者外周血单个核细胞中的 Tregs。与天然 Tregs 相比，rATG 诱导的 Tregs 表型不同，但具有相似的调节活性。rATG 可能有助于控制同种异体反应的机制。
