Infusion of clinical-grade enriched regulatory T cells delays experimental xenogeneic graft-versus-host disease
Infusion of clinical-grade enriched regulatory T cells delays experimental xenogeneic graft-versus-host disease
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DOI：
10.1111/trf.12279
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发表时间：
2014-02-01
期刊：
影响因子：
2.9
通讯作者：
Baron, Frederic
中科院分区：
文献类型：
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作者：
Hannon, Muriel;Lechanteur, Chantal;Baron, Frederic
我们研究了临床级富集的人调节性T细胞（Treg）减轻NSG小鼠中由外周血单核细胞（PBMNCs; Treg自体）输注诱导的实验性异种移植物抗宿主病（GVHD）的能力，以及证实当单独输注时它们不能诱导异种GVHD。研究设计和方法在六个独立实验中，用六个不同的健康志愿者供体，使用两步程序（同时去除CD 8和CD 19，然后进行CD 25阳性选择），用细胞分离系统（CliniMACS，Miltenyi Biotec GmbH）从外周血单采产物中分离人Treg。经亚致死（2.5戈伊）辐照的NSG小鼠静脉内（IV）给予2 × 10（6）个细胞分离术（PBMNC）产物细胞，不含Treg（PBMNC组）或含1 × 10（6）个Treg（PBMNC + Treg组），而其它NSG小鼠仅接受2 × 10（6）个富集的Treg（也在IV中; Treg组）。结果前五个程序成功获得了相对纯的Treg群体（定义为>50%），而第六个程序由于技术问题而失败（Treg纯度，42%）。在前五个实验中，Treg共输注显著延迟了异种GVHD的死亡（p < 0.0001），但在第六个实验中没有。重要的是，单独给予富集Treg的小鼠（Treg组）均未经历GVHD的临床体征，而有趣的是，移植后26天在这些小鼠中发现的CD 4+细胞主要是常规T细胞（人CD 4+总细胞中的中值CD 25 + FoxP 3+细胞在脾、骨髓和血液中分别仅为2.1、3.1和12.2%）。结论在小鼠模型中，临床级富集Treg的输注延迟了异种GVHD的发生，而不诱导毒性。
Background We investigated the ability of clinical-grade enriched human regulatory T cells (Treg) to attenuate experimental xenogeneic graft-versus-host disease (GVHD) induced by peripheral blood mononuclear cells (PBMNCs; autologous to Treg) infusion in NSG mice, as well as verified their inability to induce xenogeneic GVHD when infused alone. Study Design and Methods Human Treg were isolated from peripheral blood apheresis products with a cell separation system (CliniMACS, Miltenyi Biotec GmbH) using a two-step procedure (simultaneous CD8 and CD19 depletion followed by CD25-positive selection) in six independent experiments with six different healthy volunteer donors. Sublethally (2.5 Gy) irradiated NSG mice were given 2 x 10(6) cytapheresis (PBMNC) product cells intravenously (IV) without (PBMNC group) or with 1 x 10(6) Treg (PBMNC + Treg group), while other NSG mice received 2 x 10(6) enriched Treg alone (also in IV; Treg group). Results The first five procedures were successful at obtaining a relatively pure Treg population (defined as >50%), while the sixth procedure, due to a technical problem, was not (Treg purity, 42%). Treg cotransfusion significantly delayed death from xenogeneic GVHD in the first five experiments, (p < 0.0001) but not in the sixth experiment. Importantly, none of the mice given enriched Treg alone (Treg group) experienced clinical signs of GVHD, while, interestingly, the CD4+ cells found in these mice 26 days after transplantation were mainly conventional T cells (median CD25+FoxP3+ cells among human CD4+ total cells were only 2.1, 3.1, and 12.2% in spleen, marrow, and blood, respectively). Conclusions Infusion of clinical-grade enriched Treg delayed the occurrence of xenogeneic GVHD without inducing toxicity in this murine model.