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作者应用12-0-+四酰佛波醇13-乙酸(TPA)和钙离子载体A23187激活灰黄层淋巴细胞以大规模制备并纯化人白细胞介素2(IL-2)。方法如下:(1)TPA和A23187溶解于无水乙醇,浓度分别为20μg/ml及100μg/ml。(2)每一灰黄层得自献血者24小时内450ml全血,每次试验共需50个灰黄层;体积为1600～1800ml,约含1×10~(11)白细胞。(3)用血细胞洗脱仪去除灰黄层内的红细胞,11次实验去除90%的红细胞,并可使淋巴细胞与粒细胞的比率有所升高。从而解决了应用灰黄层作为大规模生产IL-2的人淋巴细胞的来源。随后用羟乙基淀粉(HES)梯度离心进一
The authors used gray-scale lymphocytes activated by 12-0- + tetraacyl phorbol 13-acetic acid (TPA) and calcium ionophore A23187 to prepare and purify human interleukin-2 (IL-2) on a large scale. Methods are as follows: (1) TPA and A23187 were dissolved in absolute ethanol at concentrations of 20 μg / ml and 100 μg / ml, respectively. (2) Each buffy coat was obtained from 450 ml of whole blood within 24 hours of blood donation. Each trial required a total of 50 greyish layers; the volume was 1600-1800 ml, containing about 1 x 10-11 white blood cells. (3) using blood cell eluent to remove the gliacyte within the gray layer, 11 experiments to remove 90% of the red blood cells, and can make the ratio of lymphocytes and granulocytes increased. Thus solving the problem of applying ghosting as a source of human lymphocytes for large-scale production of IL-2. Subsequently, a gradient was applied using a hydroxyethyl starch (HES) gradient