Pool E J, Johaar G, James S, Petersen I, Bouic P
Western Province Blood Transfusion Service, Parow, South Africa.
J Immunoassay. 1999 Feb-May;20(1-2):79-89. doi: 10.1080/01971529909349315.
Purified E.coli endotoxin, Gram negative bacteria and Gram positive bacteria induce IL-6 secretion by whole blood cultures (WBC's). Polymyxin B at concentrations greater than 2 U/ml completely inhibits IL-6 secretion caused by 10 EU/ml of endotoxin. Polymyxin B has no effect on IL-6 secretion by WBC's in the absence of endotoxin. The inhibition of endotoxin induced IL-6 secretion is Polymyxin B concentration dependent at concentrations less than 1 U/ml. IL-6 induction caused by E.coli is only partially inactivated by 8 U/ml Polymyxin B. Polymyxin B has no effect on IL-6 secretion caused by B.subtilis. Two pyrogenic batches of human serum albumin (HSA), as tested by the rabbit assay for pyrogens, were also investigated. Polymyxin B at 4 U/ml inhibits less than 40 % of IL-6 secretion caused by these pyrogenic HSA batches. All the endotoxin activity in HSA samples spiked with purified endotoxin is inhibited by Polymyxin B indicating that HSA does not protect endotoxin against Polymyxin B inhibition. These results indicate that the pyrogenicity of these HSA batches are caused by Polymyxin B inhibitable and non-inhibitable fractions. This study shows that pyrogenic substances other than endotoxin can contaminate batches of pharmaceutical products and that results obtained using the Limulus Amoebocyte Lysate (LAL) assay does not necessarily indicate the pyrogenic status of pharmaceutical products. The WBC assay for pyrogens, having a broader sensitivity range than the LAL assay, is a better indicator of the pyrogenic status of pharmaceutical products.
纯化的大肠杆菌内毒素、革兰氏阴性菌和革兰氏阳性菌可诱导全血培养物（WBC）分泌白细胞介素-6（IL-6）。浓度大于2 U/ml的多粘菌素B可完全抑制由10 EU/ml内毒素引起的IL-6分泌。在没有内毒素的情况下，多粘菌素B对WBC分泌IL-6没有影响。在浓度小于1 U/ml时，多粘菌素B对内毒素诱导的IL-6分泌的抑制作用呈浓度依赖性。8 U/ml的多粘菌素B只能部分灭活由大肠杆菌引起的IL-6诱导。多粘菌素B对枯草芽孢杆菌引起的IL-6分泌没有影响。还对两批经兔热源试验检测为致热的人血清白蛋白（HSA）进行了研究。4 U/ml的多粘菌素B抑制这些致热HSA批次引起的IL-6分泌不到40%。用纯化内毒素加标的HSA样品中的所有内毒素活性均被多粘菌素B抑制，这表明HSA不能保护内毒素免受多粘菌素B的抑制。这些结果表明，这些HSA批次的致热性是由多粘菌素B可抑制和不可抑制的部分引起的。本研究表明，除内毒素外的致热物质可污染药品批次，并且使用鲎试剂法（LAL）测定获得的结果不一定表明药品的致热状态。与LAL测定相比，WBC热源测定具有更宽的灵敏度范围，是药品致热状态的更好指标。
