Viselli S M, Mastro A M
Department of Molecualr and Cell Biology, Pennsylvania State University, University Park 16802.
J Immunol Methods. 1989 Dec 20;125(1-2):115-24. doi: 10.1016/0022-1759(89)90084-7.
In this report we present the use of cell blotting for the detection of interleukin-2 (IL-2)-producing lymphocytes. This is a rapid and sensitive immunochemical method analogous to Western blotting of proteins. When combined with image analysis one can determine the percentages of IL-2 positive cells as well as quantitate the amount of IL-2 surrounding each cell. When bovine lymph node cells (bLNC) were stimulated with the combination of concanavalin A (ConA) and 12-O-tetradecanoylphorbol-13-acetate (TPA) for 24 h, 46.4 +/- 0.6% stained positive for IL-2 and, on average, each cell produced 0.92 +/- 0.6 pg of IL-2 in 24 h. Phytohemagglutinin (PHA) and TPA-stimulated human peripheral blood mononuclear cells (hPBMC) produced approximately the same amount. 0.86 +/- 0.4 pg of IL-2 per cell in 24 h; 45.6 +/- 3.6% stained positive for IL-2.
在本报告中，我们介绍了使用细胞印迹法检测产生白细胞介素-2（IL-2）的淋巴细胞。这是一种快速且灵敏的免疫化学方法，类似于蛋白质的蛋白质印迹法。与图像分析相结合时，可以确定IL-2阳性细胞的百分比，并对每个细胞周围的IL-2量进行定量。当用伴刀豆球蛋白A（ConA）和12-O-十四酰佛波醇-13-乙酸酯（TPA）联合刺激牛淋巴结细胞（bLNC）24小时时，46.4±0.6%的细胞对IL-2染色呈阳性，平均每个细胞在24小时内产生0.92±0.6皮克的IL-2。植物血凝素（PHA）和TPA刺激的人外周血单个核细胞（hPBMC）产生的量大致相同，每个细胞在24小时内产生0.86±0.4皮克的IL-2；45.6±3.6%的细胞对IL-2染色呈阳性。
