Tremblay Mikaela M, Houtman Jon C D
Department of Microbiology, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, United States.
Department of Microbiology, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, United States.
J Immunol Methods. 2015 Jan;416:137-45. doi: 10.1016/j.jim.2014.11.009. Epub 2014 Nov 25.
Buffy coats are the most common method for the acquisition of activated primary human T cells for research or clinical applications, but recently leukocyte reduction system (LRS) cones have emerged as a viable source for these cells. In this study, we determined if activated human T cells derived from buffy coats or LRS cones had different functionality. No changes in the expression of surface receptors were observed except for a significant increase in CD44 expression on T cells isolated from LRS cones. LRS cone-derived T cells trended towards higher receptor-mediated cytokine production and had significantly increased donor-to-donor variability in IFN-γ production. TCR-induced ERK1/ERK2 and AKT phosphorylation was also increased in T cells isolated from LRS cones. In conclusion, LRS cones are an excellent source of T cells for clinical and research applications, but these cells have subtle functional differences from T cells isolated using standard buffy coats.
血沉棕黄层是获取活化的原代人T细胞用于研究或临床应用的最常用方法,但最近白细胞滤除系统(LRS)滤器已成为这些细胞的一个可行来源。在本研究中,我们确定了源自血沉棕黄层或LRS滤器的活化人T细胞是否具有不同的功能。除了从LRS滤器分离的T细胞上CD44表达显著增加外,未观察到表面受体表达的变化。源自LRS滤器的T细胞趋向于产生更高的受体介导的细胞因子,并且在IFN-γ产生方面供体间变异性显著增加。从LRS滤器分离的T细胞中TCR诱导的ERK1/ERK2和AKT磷酸化也增加。总之,LRS滤器是临床和研究应用中T细胞的优良来源,但这些细胞与使用标准血沉棕黄层分离的T细胞存在细微的功能差异。