Department of Pediatrics, Division of Hematology and Oncology, Faculty of Medicine Ramathibodi Hospital.
Department of Pediatrics, Pediatric Translational Research Unit, Faculty of Medicine Ramathibodi Hospital.
J Immunother. 2019 May;42(4):110-118. doi: 10.1097/CJI.0000000000000262.
The third-party umbilical cord blood (UCB)-derived regulatory T cells (Treg) are an alternative to donor-derived Treg as cellular therapy of graft-versus-host disease following hematopoietic stem cell transplantation. However, their suppressive characteristics against autologous and allogeneic T effector cells (Teff) have rarely been documented. The exact role of UCB-Treg in hematologic malignancies is also uncertain. Here, we investigated the direct effects of UCB-Treg on the proliferation of autologous Teff, as compared with allogeneic Teff, and also determined cellular fates of lymphoblasts after UCB-Treg co-culture. UCB-Treg were isolated from 8 UCB samples using 2-step immunomagnetic bead sorting. After 10-day ex vivo expansion, up to 60-fold increase in cell number with 76.7%±4.9% of CD4CD25CD127FoxP UCB-Treg was obtained. Further characterization showed that ex vivo-expanded UCB-Treg contained a higher proportion of CD95CD45RACCR4Treg-B subpopulation compared with the CD95CD45RACCR4Treg-A subpopulation (13.0%±4.8% vs. 0.8%±0.7%; P<0.05), along with the detecting of substantial amounts of secretory IL-10 (57.7±17.8 pg/mL) and TGF-β1 (196.5±29.7 pg/mL) in culture supernatants. After 4 days co-culture with UCB-Treg (at the ratio of 1:1), the proliferation of autologous and allogeneic Teff was decreased comparably (43.6%±17.5% vs. 37.6±17.7%; P=0.437). Suppression was independent of HLA-A, B, and DRB1 compatibility between UCB-Treg and Teff. UCB-Treg co-culture with various lymphoblasts showed proliferative suppression of Jurkat T lymphoblasts (45.4%±20.5% at the ratio of 1:1), but not Namalwa and Raji B lymphoblasts. All lymphoblasts had no significant cell apoptosis or death after co-culture. In conclusion, the ex vivo-expanded UCB-Treg had no difference in autologous and allogeneic Teff suppression. UCB-Treg therapy in patients with graft-versus-host disease who have a primary disease of T-cell leukemia may have additional benefits in the prevention of relapsed disease.
第三方脐带血（UCB）来源的调节性 T 细胞（Treg）是供体来源的 Treg 作为造血干细胞移植后移植物抗宿主病的细胞治疗的替代物。然而，它们对自体和同种异体效应 T 细胞（Teff）的抑制特性很少有文献记载。UCB-Treg 在血液恶性肿瘤中的确切作用也不确定。在这里，我们研究了 UCB-Treg 对自体 Teff 增殖的直接影响，与同种异体 Teff 进行了比较，还确定了 UCB-Treg 共培养后淋巴母细胞的细胞命运。使用两步免疫磁珠分选从 8 个 UCB 样本中分离 UCB-Treg。经过 10 天的体外扩增，细胞数量增加了 60 倍，获得了 76.7%±4.9%的 CD4CD25CD127FoxP UCB-Treg。进一步的特征表明，与 CD95CD45RACCR4Treg-A 亚群相比，体外扩增的 UCB-Treg 含有更高比例的 CD95CD45RACCR4Treg-B 亚群（13.0%±4.8%比 0.8%±0.7%；P<0.05），同时培养上清液中检测到大量分泌的 IL-10（57.7±17.8 pg/mL）和 TGF-β1（196.5±29.7 pg/mL）。与 UCB-Treg（比例为 1:1）共培养 4 天后，自体和同种异体 Teff 的增殖受到类似程度的抑制（43.6%±17.5%比 37.6±17.7%；P=0.437）。抑制与 UCB-Treg 和 Teff 之间 HLA-A、B 和 DRB1 相容性无关。UCB-Treg 与各种淋巴母细胞共培养显示 Jurkat T 淋巴母细胞增殖受到抑制（比例为 1:1 时为 45.4%±20.5%），但 Namalwa 和 Raji B 淋巴母细胞不受影响。所有淋巴母细胞在共培养后均无明显细胞凋亡或死亡。总之，体外扩增的 UCB-Treg 对自体和同种异体 Teff 的抑制作用没有差异。在原发性 T 细胞白血病患者中，UCB-Treg 治疗移植物抗宿主病可能在预防疾病复发方面具有额外的益处。
