Larrick J W, Brindley L, Doyle M V
J Immunol Methods. 1985 May 10;79(1):39-45. doi: 10.1016/0022-1759(85)90389-8.
A 24 h, highly sensitive assay for detection of interleukin 1 (IL-1) is described. A thioguanine-resistant mutant of the murine lymphoma cell line LBRM 33 was selected (LBRM TG6). When this cell line was incubated with low concentrations of PHA and IL-1 it produced interleukin 2 (IL-2). IL-2-dependent HT2 cells were co-cultured with the LBRM cells to measure the released IL-2. Prior to addition of tritiated thymidine to the co-culture, hypoxanthine and azaserine were added to metabolically block DNA synthesis by LBRM TG6 cells. This resulted in a sensitive, short term assay requiring minimal technical manipulations and characterized by a high signal to noise ratio.
描述了一种用于检测白细胞介素1(IL-1)的24小时高灵敏度检测方法。选择了鼠淋巴瘤细胞系LBRM 33的硫鸟嘌呤抗性突变体(LBRM TG6)。当该细胞系与低浓度的PHA和IL-1一起孵育时,它会产生白细胞介素2(IL-2)。将依赖IL-2的HT2细胞与LBRM细胞共培养以测量释放的IL-2。在向共培养物中添加氚标记的胸腺嘧啶核苷之前,添加次黄嘌呤和重氮丝氨酸以代谢性阻断LBRM TG6细胞的DNA合成。这产生了一种灵敏的短期检测方法,所需的技术操作最少,并且具有高信噪比。