Landwehr-Kenzel S, Issa F, Luu S-H, Schmück M, Lei H, Zobel A, Thiel A, Babel N, Wood K, Volk H-D, Reinke P
Berlin-Brandenburg Center for Regenerative Therapies, Charité University Medicine Berlin, Berlin, Germany; Berlin-Brandenburg School for Regenerative Therapies, Charité University Medicine Berlin, Berlin, Germany; Department of Pediatric Pulmonology and Immunology, Charité University Medicine Berlin, Berlin, Germany.
Am J Transplant. 2014 Mar;14(3):594-606. doi: 10.1111/ajt.12629. Epub 2014 Jan 27.
The adoptive transfer of natural regulatory T cells (nTreg) is a new option to reshape undesired immune reactivity in autoimmunity and transplantation toward "tolerance." The first clinical trials using adoptive transfer of polyclonal nTreg demonstrated safety and hints of efficacy. However, the low frequencies of antigen-specific cells among the pool of polyclonal nTreg and their broad antigen nonspecific suppression are limitations of this approach regarding efficacy and safety. Recently, the isolation and expansion of (allo)antigen-specific nTreg have successfully been achieved by using Treg-specific activation markers but the yield is relatively low. Here, we describe a novel good manufacturing practice (GMP)-compatible expansion protocol of alloantigen-specific nTreg based on the stimulation of nTreg by allogeneic activated B cells. Their functionality and specificity are superior compared to polyclonal nTreg both in vitro and in vivo. Employing an allogeneic B cell bank, designed to cover the majority of HLA types, allows fast GMP-compliant manufacturing for donor-specific nTreg for clinical application in organ and stem cell transplantation. TCR repertoire analyses by next generation sequencing revealed impressive expansion by several log-steps of even very low-abundance alloantigen-specific nTreg clones. This novel method offers a simple approach for expanding antigen-specific nTreg and is characterized by high replicability and easy transferability to full GMP standards.
天然调节性T细胞（nTreg）的过继性转移是一种重塑自身免疫和移植中不良免疫反应以实现“耐受”的新方法。首批使用多克隆nTreg过继性转移的临床试验证明了其安全性和疗效迹象。然而，多克隆nTreg库中抗原特异性细胞频率较低及其广泛的抗原非特异性抑制作用，在疗效和安全性方面限制了这种方法。最近，通过使用Treg特异性激活标记物，已成功实现（同种异体）抗原特异性nTreg的分离和扩增，但产量相对较低。在此，我们描述了一种基于同种异体活化B细胞刺激的、与药品生产质量管理规范（GMP）兼容的新型同种异体抗原特异性nTreg扩增方案。在体外和体内，其功能和特异性均优于多克隆nTreg。利用旨在涵盖大多数HLA类型的同种异体B细胞库，能够快速生产符合GMP标准的供体特异性nTreg，用于器官和干细胞移植的临床应用。通过下一代测序进行的TCR库分析显示，即使是非常低丰度的同种异体抗原特异性nTreg克隆也能实现几个对数级的显著扩增。这种新方法为扩增抗原特异性nTreg提供了一种简单途径，具有高重复性且易于转换为完全符合GMP标准的方法。
