Tran Dat Q, Andersson John, Hardwick Donna, Bebris Lolita, Illei Gabor G, Shevach Ethan M
Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 20892, USA.
Blood. 2009 May 21;113(21):5125-33. doi: 10.1182/blood-2009-01-199950. Epub 2009 Mar 18.
Although adoptive transfer of regulatory T cells (Foxp3(+) Tregs) has proven to be efficacious in the prevention and treatment of autoimmune diseases and graft-versus-host disease in rodents, a major obstacle for the use of Treg immunotherapy in humans is the difficulty of obtaining a highly purified preparation after ex vivo expansion. We have identified latency-associated peptide (LAP) and IL-1 receptor type I and II (CD121a/CD121b) as unique cell-surface markers that distinguish activated Tregs from activated FOXP3(-) and FOXP3(+) non-Tregs. We show that it is feasible to sort expanded FOXP3(+) Tregs from non-Tregs with the use of techniques for magnetic bead cell separation based on expression of these 3 markers. After separation, the final product contains greater than 90% fully functional FOXP3(+) Tregs. This novel protocol should facilitate the purification of Tregs for both cell-based therapies as well as detailed studies of human Treg function in health and disease.
尽管调节性T细胞（Foxp3(+) Tregs）的过继转移已被证明在预防和治疗啮齿动物自身免疫性疾病及移植物抗宿主病方面有效，但在人类中使用Treg免疫疗法的一个主要障碍是离体扩增后难以获得高度纯化的制剂。我们已确定潜伏相关肽（LAP）以及I型和II型白细胞介素-1受体（CD121a/CD121b）为独特的细胞表面标志物，可将活化的Tregs与活化的FOXP3(-)和FOXP3(+)非Tregs区分开来。我们表明，利用基于这3种标志物表达的磁珠细胞分离技术从非Tregs中分选扩增的FOXP3(+) Tregs是可行的。分离后，最终产物含有超过90%的功能完全正常的FOXP3(+) Tregs。这种新方案应有助于纯化Tregs，用于基于细胞的治疗以及对人类Treg在健康和疾病中的功能进行详细研究。
