ISOLATION AND CHARACTERIZATION OF AN LIPOPOLYSACCHARIDE (LPS)-RESISTANT GENE FROM AN LPS-REISITANT MACROPHAGE-LIKE CELL LINE

建立了一种评价脂多糖抗性的实验方法：10 μ g/ml环己亚胺和10 μ g/ml脂多糖均可诱导脂多糖敏感的巨噬细胞样细胞JA-4 （J774.1亚系）在37℃孵育4 h后产生严重的细胞损伤。而对LPS具有高度抗性的LPS1916细胞系，即使在环己亚胺和1杯/毫升LPS孵育后，也没有受到损伤。与细胞生长试验或集落形成试验相比，该方法对lps抗性的估计敏感且快速。磷酸钙法、电穿孔法和脂肪转染法均未成功转染JA-4细胞。由于最近有其他实验室的报道表明转染了荧光素酶基因后，RAW 264.7细胞表达了荧光素酶基因，因此我们正在用另一种对LPS敏感的巨噬细胞样细胞系RAW 264.7作为LPS1916和JA-4细胞的cDNAa的受体进行初步实验。

An assay method was newly developped for evaluation of LPS-resistance : Both 10 mug/ml cycloheximide and 10 ng/ml LPS induced the severe cell damages in an LPS-sensitive macrophage-like cell, JA-4, a subline of J774.1, andafter incubation at 37ﾟC for 4th.However, LPS1916 cell line, a highly resistant cell line to LPS,was not damaged at all, even after incubation with cycloheximide and 1 mug/ml LPS.This method was sensitive and rapid for estimation of LPS-resistance, as compared with a cell growth assay or a colony-formation assay.Trials to transfect DNA to JA-4 cells were not successful with calcium phosphate method, electro-poration method or lipofection method.A preliminary experiment is being performed by using another macrophage-like cell line, RAW 264.7, which is sensitive to LPS,as a receipient of the cDNAa from LPS1916 as well as JA-4 cells, because a recent report from other laboratory suggested that RAW 264.7 cells expressed luciferase gene after transfection of the gene.