Novel application of a whole blood intracellular cytokine detection assay to quantitate specific T-cell frequency in field studies
Novel application of a whole blood intracellular cytokine detection assay to quantitate specific T-cell frequency in field studies
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DOI：
10.1016/j.jim.2004.06.010
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发表时间：
2004-08-01
影响因子：
2.2
通讯作者：
Kaplan, G
中科院分区：
文献类型：
--
作者：
Hanekom, WA;Hughes, J;Kaplan, G
我们优化了全血细胞内细胞因子测定，以定量来自发展中国家婴儿的小体积全血中特异性CD 4（+）和CD 8（+）T细胞的频率。该测定分两步进行。首先，在特异性抗原存在下刺激全血6-18小时，以冷冻保存固定的白色细胞结束。这些刺激步骤经过专门调整，使其在农村、发展中国家的实地环境中实用且可靠。随后，在更有资源的环境中，通过流式细胞术检测产生干扰素-γ的CD 4（+）或CD 8（+）T细胞。该方法被证明是敏感和特异性检测分枝杆菌特异性免疫接种卡介苗（BCG）后10周的新生儿从农村现场。(C)2004 Elsevier B. V.保留所有权利。
We optimized a whole blood intracellular cytokine assay to quantitate the frequency of specific CD4(+) and CD8(+) T cells in small volumes of whole blood from infants from developing countries. The assay is performed in two steps. First, whole blood is stimulated in the presence of specific antigens for 6-18 h, ending with cryopreservation of fixed white cells. These stimulation steps were specifically adapted to be practical and reliable in a rural, developing country field setting. Later, in a more resourceful setting, interferon-gamma producing CD4(+) or CD8(+) T cells are detected by flow cytometry. The assay proved sensitive and specific for detecting mycobacteria-specific immunity 10 weeks after Bacillus Calmette-Guerin (BCG) vaccination of newborns from a rural field site. (C) 2004 Elsevier B.V. All rights reserved.