Vandenabeele P, Declercq W, Libert C, Fiers W
Laboratory of Molecular Biology, State University of Ghent, Belgium.
J Immunol Methods. 1990 Dec 31;135(1-2):25-32. doi: 10.1016/0022-1759(90)90252-q.
The IL-1-dependent proliferation of RPMI 1788, a human EBV-transformed cell line, was used to develop a biological assay system for IL-1. Preparations of rhIL-1 alpha and rhIL-1 beta, as well as rmIL-1 beta exhibited a specific biological activity (50% of the maximal response) between 5.8 x 10(8) and 8.6 x 10(8) U/mg. Remarkably, a 3-5-fold reduced specific biological activity was noticed for rm-IL-1 alpha, viz. 1.7 x 10(8) U/mg. The IL-1-dependent proliferation of RPMI 1788 cells was compared with other IL-1 test systems, such as the IL-1-mediated induction of IL-2 in EL4-NOB-1, LBRM-33-1A5 and thymocytes, and the IL-1-driven induction of cytotoxic activity by PC60 cells, the so-called CIA assay. The cytokine-dependent growth of RPMI 1788 cells is highly specific for IL-1, and no other cytokine tested induced a proliferative response. The presence of high concentrations of rmTNF, rhTNF or rhIL-6 did not interfere with the quantification of IL-1. Additionally, we evaluated the detection of IL-1 in the presence of mitogens, phorbol ester or calcium ionophore, as well as the determination of IL-1 in serum and PF samples of human and murine origin.
人EBV转化细胞系RPMI 1788依赖白细胞介素-1（IL-1）的增殖被用于开发一种IL-1的生物检测系统。重组人IL-1α、重组人IL-1β以及重组小鼠IL-1β制剂在5.8×10⁸至8.6×10⁸U/mg之间表现出特定的生物活性（最大反应的50%）。值得注意的是，重组小鼠IL-1α的特定生物活性降低了3至5倍，即1.7×10⁸U/mg。将RPMI 1788细胞依赖IL-1的增殖与其他IL-1检测系统进行了比较，如IL-1介导的EL4-NOB-1、LBRM-33-1A5和胸腺细胞中IL-2的诱导，以及PC60细胞由IL-1驱动的细胞毒活性诱导，即所谓的CIA检测。RPMI 1788细胞依赖细胞因子的生长对IL-1具有高度特异性，测试的其他细胞因子均未诱导增殖反应。高浓度的重组小鼠肿瘤坏死因子（rmTNF）、重组人肿瘤坏死因子（rhTNF）或重组人白细胞介素-6（rhIL-6）的存在不干扰IL-1的定量。此外，我们评估了在有丝分裂原、佛波酯或钙离子载体存在的情况下IL-1的检测，以及人源和鼠源血清及PF样品中IL-1的测定。
