内生致热原制备的实验研究

An experimental method for endogenous pyrogen preparation from monocytes of rabbits' blood

本研究应用Ficoll-Hypaque分离技术,从离体兔血液获得单核细胞,用生理盐水配成含单核细胞0.65～1.0×10~6cell/ml的悬液。以精制大肠杆菌内毒素激活,37—38℃振荡孵育10小时。离心收集上清液进行超滤。收集液中含单核细胞内生致热原(monocytic endogenous pyrogen)。实验证明:通过Ficoll-Hypaque分离技术可获得高浓度的单核细胞内生致热原,该内生致热原给动物静脉注射引起明显的发热。

In order to obtain monocytic endogenous pyrogen, Ficoll—Hypaque's separation technique was used in this paper. We obtained mononuclear cells from rabbits' blood in vitro and added endotoxin free saline to make suspension in the concentration of 0.65×10~6/mL. Endogenous pyrogen was produced from monocytes by stimulation of purified endotoxin of colon bacilli and incubation in saline at 37-38℃ stirred and shaked continuously for 10 hr. There is EP in collecting fluid after centrifugation and ultrafiitration.The result obtained shows:1. Employing Ficoll-Hypaque's separation technique, high concentration monocytic endogenous pyrogen can be obtained.2. Monocytic endogenous pyrogen have marked febrile effect.