Bresatz Suzanne, Sadlon Tim, Millard Debrah, Zola Heddy, Barry Simon C
Discipline of Paediatrics, Adelaide, Australia.
J Immunol Methods. 2007 Oct 31;327(1-2):53-62. doi: 10.1016/j.jim.2007.06.006. Epub 2007 Jul 17.
Regulatory T cells (Treg) have recently come to the fore in studies of immune regulation, particularly in autoimmune disease and cancer. While there appear to be several distinct subsets of T cells with regulatory function, a population described as natural Treg and characterized by expression of the transcription factor FOXP3 has attracted particular interest. These cells can be enriched using the surface markers CD4 and CD25, and cord blood is a convenient source of CD25+ Treg. We present detailed protocols for the enrichment of Treg from cord blood using CD25 and a magnetic bead procedure, yielding populations >80% positive for CD25 and 50-65% FOXP3 positive. This enrichment can be followed by a second magnetic bead or a flow sorting step, yielding >95% CD25 and >65% FOXP3 positive populations. Protocols are presented for propagation of these cells in culture (yielding >80% FOXP3 positive cells) and for their phenotypic and functional characterization.
调节性T细胞（Treg）最近在免疫调节研究中备受关注，尤其是在自身免疫性疾病和癌症研究方面。虽然似乎存在几个具有调节功能的不同T细胞亚群，但一种被称为天然Treg并以转录因子FOXP3表达为特征的细胞群体引起了特别关注。这些细胞可以使用表面标志物CD4和CD25进行富集，而脐带血是CD25+ Treg的便捷来源。我们提供了使用CD25和磁珠程序从脐带血中富集Treg的详细方案，得到的细胞群体中CD25阳性率>80%，FOXP3阳性率为50 - 65%。这种富集之后可以进行第二次磁珠或流式分选步骤，得到CD25阳性率>95%且FOXP3阳性率>65%的细胞群体。文中还给出了这些细胞在培养中增殖（得到>80% FOXP3阳性细胞）及其表型和功能特征鉴定的方案。
