一种高度敏感的改良的IL-1检测方法

A highly sensitive, modified assay for Interleukin-1

采用CTLL-2检测IL-1诱导小鼠胸腺瘤细胞系EL_4产生IL-2活性,建立了一种间接检测IL-1的方法。通过对多种影响因素进行探讨,选择了较适的诱导血清浓度,细胞浓度,诱导时间和转移诱导上清稀释度,从而使检测IL-1的敏感性达10^(-3)～10^(-4)U/ml(2.5～25×10^(-4)Pg/ml,约为小鼠胸腺细胞检测方法的10~2～10~3倍),整个检测流程可在40小时内完成。若用CTLL-2和EL_4细胞同时培养的一步法检测,敏感性约为10^(-1)U/ml,但30小时内可完成检测。此法不受高浓度rHuTNF-α、rHuTNF-β、rHuIL-6干扰,IL-2、ConA、PHA、LPS和SEB对检测系统只有较弱的影响,但A23187可明显地干扰检测系统。因此,本方法可用于基础和临床研究过程中不同来源的IL-1生物学活性检测。

In this paper, we describe a highly sensitive, modified assay which takes advantage of the capacity of a mouse thymoma cell line EL4 subclone to produce high concentration of IL-2 upon stimulation by interleukin I (IL-1).The assay was generally performed in 2 stages which included IL-1-dependent IL-2 production and IL-2 biological activity assay, and took about 40 hours to complete. The sensitivity of this assay was got to be 10^(-2) ～10^(-4)U/ml (2.5～25×10^(-4)pg/ml, about 10~2～10~3 times more sensitive than that of the mouse thymocyte co-stimulated assay) by selecting the optimal serum concentration, EL 4 cell density, IL-1-induced time and EL4 supernatant dilution. The rapid one step assay was performed by co-culturing the EL4 cells with CTLL-2 cells. This rapid assay could be completed within 30 hours with sensitivity of 10^(-1)U/ml. The assay was not affected by high concentrations of rHuTNF- α, rHuTNF- β or rHuIL-6,and was only slightly affected by high concentrations of rHuIL-2, ConA, PHA,LPS and SEB, and much affected by A23187.