A simple and rapid magnetic bead separation technique for the isolation of tetramer-positive virus-specific CD8 T cells.
A simple and rapid magnetic bead separation technique for the isolation of tetramer-positive virus-specific CD8 T cells.
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一种简单快速的磁珠分离技术，用于分离四聚体阳性病毒特异性 CD8 T 细胞。
DOI：
10.1097/00002030-200104130-00024
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Nixon,DF
中科院分区：
文献类型：
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作者：
McDermott,AB;Spiegel,HM;Irsch,J;Ogg,GS;Nixon,DF
最近关于CD8T细胞亚群内功能损害的报道[1-5]将注意力集中在对功能活动的定量评估上。然而，抗原特异性CD8T细胞的扩增和繁殖通常需要广泛的组织培养操作和频繁的再刺激。一种可以选择性地识别和扩大抗原特异性CD8群体的方法在这些定量评估中将是有价值的。四聚体阳性病毒特异性T细胞的流式细胞仪或人类白细胞抗原(HL A)珠分选已被证明是有价值的[6，7]；但它需要流式细胞仪，技术要求高，需要绝对无菌的条件。我们开发了一种利用抗藻红蛋白包裹的MACS小球(Miltenyi Biotec，Auburn，USA)来筛选与CD8 T细胞混合的阳性藻红蛋白偶联四聚肽复合体的方法，以获得并选择丰富的特定群体进行体外培养。这种简化的方法可以导致抗原特异性T细胞群的快速扩增。
Recent reports on functional impairments within CD8 T cell subpopulations [1–5] focused attention on quantitative assessments of functional activity. However, the expansion and propagation of antigen-specific CD8 T cells often requires extensive tissue culture manipulations and frequent re-stimulations. A method that could selectively identify and expand an antigen-specific CD8 population would be of value in these quantitative assessments.Flow cytometric or human leukocyte antigen (HLA) bead sorting of tetrameric-positive virus-specific T cells has proved to be of value [6, 7]; however, it requires a flow cytometer, is technically demanding and needs conditions of absolute sterility. We have developed an assay that utilizes anti-phycoerythrin-coated MACS beads (Miltenyi Biotec, Auburn, USA) to select positively phycoerythrin-conjugated tetramer peptide complexes admixed with CD8 T cells to obtain and select enriched specific populations for in-vitro culture. This simplified method can lead to the rapid expansion of an antigen-specific T cell population.