Pedersen M R, Jensen S, Christensen J D, Hansen E W
Royal Danish School of Pharmacy, Department of Biological Sciences, Copenhagen, Denmark.
J Immunol Methods. 1995 Mar 27;180(2):159-63. doi: 10.1016/0022-1759(94)00311-j.
The B9 assay is known to be a specific and sensitive assay for the estimation of interleukin-6 activity. This assay was found to be compromised by lipopolysaccharide in concentrations > or = 40 ng lipopolysaccharide per ml. The lipopolysaccharide stimulates proliferation of the B9 cell line in a dose-dependent manner both when measuring the proliferation by thymidine incorporation and when using the MTT assay. However the LPS dose-response curve is different compared to the dose-response curve for IL-6. A sample containing 100 ng LPS/ml but no IL-6 would be estimated erroneously to contain 12 pg IL-6. The interference of lipopolysaccharide is totally abolished by the addition of polymyxin B to the samples but the addition has no effect on the IL-6 induced proliferation.
已知B9测定法是一种用于评估白细胞介素-6活性的特异性和灵敏性测定法。发现当脂多糖浓度≥40 ng/ml时,该测定法会受到影响。无论是通过胸腺嘧啶核苷掺入法测量增殖,还是使用MTT测定法,脂多糖均以剂量依赖性方式刺激B9细胞系的增殖。然而,与IL-6的剂量反应曲线相比,LPS的剂量反应曲线有所不同。一个含有100 ng LPS/ml但不含IL-6的样品会被错误地估计为含有12 pg IL-6。通过向样品中添加多粘菌素B可完全消除脂多糖的干扰,但添加多粘菌素B对IL-6诱导的增殖没有影响。