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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。
The IL-2 activity of IL-1 induced by IL-1 in mouse thymoma cell line EL_4 was detected by CTLL-2, and an indirect method for detecting IL-1 was established. Through investigating a variety of influencing factors, the appropriate inducing serum concentration, cell concentration, induction time and the dilution of transfer induced supernatant were selected so that the sensitivity of detecting IL-1 reached 10-3 ~ (-4) U / ml (2.5 ~ 25 × 10 -4 Pg / ml, which is about 10 ~ 2 ~ 10 ~ 3 times higher than that of mouse thymocyte detection method). The entire detection process can be completed in 40 hours . If CTLL-2 and EL_4 cells simultaneously cultured in one-step assay, the sensitivity of about 10 ~ (-1) U / ml, but within 30 hours to complete the test. This method is not affected by high concentrations of rHuTNF-α, rHuTNF-β, rHuIL-6, and IL-2, ConA, PHA, LPS and SEB have a weaker effect on the detection system, but A23187 can significantly interfere with the detection system. Therefore, the method can be used for the detection of IL-1 biological activity in different sources during basic and clinical research.