输注临床级别的富含调节性 T 细胞可延迟实验性异种移植物抗宿主病。

Infusion of clinical-grade enriched regulatory T cells delays experimental xenogeneic graft-versus-host disease.

机构信息

Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I3, Laboratory of Cell and Genetic Therapy, Department of Pathology, Department of Statistics, GIGA-R, Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium; de Duve Institute, Université Catholique de Louvain, Brussels, Belgium; Red Cross Transfusion Center of Liege, Liège, Belgium; Autoimmune Genetics Laboratory, University of Leuven, Leuven, Belgium.

出版信息

Transfusion. 2014 Feb;54(2):353-63. doi: 10.1111/trf.12279. Epub 2013 Jun 17.

DOI:10.1111/trf.12279

PMID:23772685

Abstract

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 × 10(6) cytapheresis (PBMNC) product cells intravenously (IV) without (PBMNC group) or with 1 × 10(6) Treg (PBMNC + Treg group), while other NSG mice received 2 × 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.

摘要

背景

我们研究了临床级富集的人调节性 T 细胞（Treg）通过输注外周血单个核细胞（PBMNC；与 Treg 自体）来减弱实验性异种移植物抗宿主病（GVHD）的能力，以及验证了当单独输注时它们不能诱导异种 GVHD 的能力。

研究设计和方法

使用细胞分离系统（Miltenyi Biotec GmbH 的 CliniMACS）从外周血单采产品中分离人 Treg，使用两步法（同时 CD8 和 CD19 耗竭，然后 CD25 阳性选择）在六个独立的实验中，每个实验使用六个不同的健康志愿者供体。亚致死剂量（2.5 Gy）照射的 NSG 小鼠静脉内（IV）给予 2×106 细胞分离术（PBMNC）产品细胞，无（PBMNC 组）或有 1×106 Treg（PBMNC+Treg 组），而其他 NSG 小鼠单独接受 2×106 富集 Treg（也在 IV 中；Treg 组）。

结果

前五步成功地获得了相对纯的 Treg 群体（定义为>50%），而第六步由于技术问题没有（Treg 纯度，42%）。Treg 共转染在前五次实验中显著延迟了异种 GVHD 的死亡（p<0.0001），但在第六次实验中没有。重要的是，单独给予富集 Treg 的小鼠（Treg 组）均未出现 GVHD 的临床症状，而有趣的是，在移植后 26 天这些小鼠中发现的 CD4+细胞主要是常规 T 细胞（人 CD4+总细胞中 CD25+FoxP3+细胞的中位数分别为脾、骨髓和血液中的 2.1%、3.1%和 12.2%）。