Center for Cell and Gene Therapy, Methodist Hospital and Texas Children's Hospital, Houston, TX, USA.
Haematologica. 2013 Apr;98(4):533-7. doi: 10.3324/haematol.2012.076430. Epub 2012 Dec 14.
The low frequency of naturally occurring regulatory T cells (nTregs) in peripheral blood and the suboptimal protocols available for their ex vivo expansion limit the development of clinical trials based on the adoptive transfer of these cells. We have, therefore, generated a simplified, robust and cost-effective platform for the large-scale expansion of nTregs using a gas permeable static culture flask (G-Rex) in compliance with Good Manufacturing Practice. More than 10(9) putative Tregs co-expressing CD25 and CD4 molecules (92 ± 5%) and FoxP3 (69 ± 19%) were obtained within 21 days of culture. Expanded Tregs showed potent regulatory activity in vitro (80 ± 13% inhibition of CD8(+) cell division) and in vivo (suppression or delay of graft-versus-host disease in a xenograft mouse model) indicating that the cost-effective and simplified production of nTregs we propose will facilitate the implementation of clinical trials based on their adoptive transfer.
外周血中天然调节性 T 细胞（nTregs）的低频以及体外扩增这些细胞的现有方案并不理想，限制了基于这些细胞过继转移的临床试验的发展。因此，我们开发了一种符合良好生产规范（Good Manufacturing Practice）的简单、稳健且具有成本效益的平台，使用透气静态培养瓶（G-Rex）来大规模扩增 nTregs。在 21 天的培养过程中，获得了超过 10（9）个表达 CD25 和 CD4 分子（92±5%）和 FoxP3（69±19%）的推定 Treg。扩增的 Treg 在体外（抑制 CD8（+）细胞分裂 80±13%）和体内（抑制或延迟异种移植小鼠模型中的移植物抗宿主病）具有强大的调节活性，表明我们提出的具有成本效益且简化的 nTreg 生产将有助于基于其过继转移的临床试验的实施。
