Kamala T
Section on T Cell Tolerance and Memory (Ghost Lab), LCMI, NIAID, NIH, Rockville Pike, Bethesda, MD 20892, USA.
Scand J Immunol. 2008 Mar;67(3):285-94. doi: 10.1111/j.1365-3083.2007.02066.x.
This study describes an immunomagnetic bead-based methodology for optimal purification of mouse CD4+ T cells. It reproducibly yields highly pure CD4+ T cells from mouse lymph nodes (95-99%) and spleen (93-96%) with no residual antigen-presenting cell (APC) function in the purified population. The recovery of the starting CD4+ T-cell population is consistently high ( > 70%) and many samples can be simultaneously processed in a short period of time. The key factors responsible for improved purity are combinations of monoclonal antibodies that were found, through trial and error, to yield T cells of maximal purity achievable by non-flow sort-based negative selection. These cocktails efficiently target unwanted cell subsets with antibodies against multiple surface markers expressed by non-CD4+ T cells. Because immunomagnetic bead-based protocols do not require the expensive and cumbersome processes required by flow sort-based purification, the methodology described here should find widespread use.
本研究描述了一种基于免疫磁珠的方法，用于优化小鼠CD4+ T细胞的纯化。该方法可重复地从小鼠淋巴结（95-99%）和脾脏（93-96%）中获得高度纯化的CD4+ T细胞，且纯化后的细胞群体中无残留抗原呈递细胞（APC）功能。起始CD4+ T细胞群体的回收率始终很高（>70%），并且可以在短时间内同时处理多个样本。提高纯度的关键因素是单克隆抗体的组合，通过反复试验发现，这些组合能够产生通过基于非流式分选的阴性选择可达到的最大纯度的T细胞。这些混合物通过针对非CD4+ T细胞表达的多种表面标志物的抗体有效地靶向不需要的细胞亚群。由于基于免疫磁珠的方案不需要基于流式分选的纯化所需昂贵且繁琐的过程，因此本文所述方法应会得到广泛应用。
