Kirikae T, Nitta T, Kirikae F, Suda Y, Kusumoto S, Qureshi N, Nakano M
Department of Microbiology, Jichi Medical School, Tochigi-ken 329-0498, Japan.
Infect Immun. 1999 Apr;67(4):1736-42. doi: 10.1128/IAI.67.4.1736-1742.1999.
Some lipopolysaccharide (LPS) preparations from S- or R-form members of the family Enterobacteriaceae and oral black-pigmented bacteria (Porphyromonas gingivalis and Prevotella intermedia) are known to activate LPS-refractory C3H/HeJ macrophages. When contaminating proteins are removed from R-form LPS of Enterobacteriaceae by repurification, however, this ability is lost. In the present study, we investigated the capacity of LPS from P. gingivalis, P. intermedia, Salmonella minnesota, and Salmonella abortusequi to induce production of tumor necrosis factor (TNF) in gamma interferon-primed C3H/HeJ macrophages before and after repurification. P. abortusequi S-LPS was fractionated by centrifugal partition chromatography into two LPS forms: SL-LPS, having homologous long O-polysaccharide chains, and SS-LPS having short oligosaccharide chains. Prior to repurification, all LPS forms except SL-LPS induced TNF production in both C3H/HeJ and C3H/HeN macrophages. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that repurification removed contaminating protein from the preparations, and repurified SS-LPS and S. minnesota Ra-LPS no longer stimulated TNF production in C3H/HeJ macrophages, although C3H/HeN macrophages remained responsive. In contrast, repurified oral bacterial LPS retained the capacity to induce TNF production in C3H/HeJ macrophages. Oral bacterial LPS preparations also were not antagonized by excess inactive, repurified SL-LPS; Ra-LPS; Rhodobacter sphaeroides lipid A, a competitive LPS antagonist, or paclitaxel, an LPS agonist, and they were comparatively resistant to polymyxin B treatment. Nevertheless, oral bacterial LPS was less toxic to D-galactosamine-treated C3H/HeN mice than was LPS from Salmonella. These findings indicate that the active molecule(s) and mode of action of LPS from P. gingivalis and P. intermedia are quite different from those of LPS from Salmonella.
已知一些来自肠杆菌科S型或R型成员以及口腔产黑色素菌（牙龈卟啉单胞菌和中间普氏菌）的脂多糖（LPS）制剂可激活对LPS不敏感的C3H/HeJ巨噬细胞。然而，当通过再纯化从肠杆菌科的R型LPS中去除污染蛋白时，这种能力就会丧失。在本研究中，我们调查了牙龈卟啉单胞菌、中间普氏菌、明尼苏达沙门氏菌和马流产沙门氏菌的LPS在再纯化前后，在γ干扰素预处理的C3H/HeJ巨噬细胞中诱导肿瘤坏死因子（TNF）产生的能力。马流产沙门氏菌S-LPS通过离心分配色谱法分离为两种LPS形式：具有同源长O-多糖链的SL-LPS和具有短寡糖链的SS-LPS。在再纯化之前，除SL-LPS外的所有LPS形式在C3H/HeJ和C3H/HeN巨噬细胞中均诱导TNF产生。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示，再纯化去除了制剂中的污染蛋白，再纯化的SS-LPS和明尼苏达沙门氏菌Ra-LPS不再刺激C3H/HeJ巨噬细胞产生TNF，尽管C3H/HeN巨噬细胞仍有反应。相反，再纯化的口腔细菌LPS保留了在C3H/HeJ巨噬细胞中诱导TNF产生的能力。口腔细菌LPS制剂也不会被过量的无活性、再纯化的SL-LPS、Ra-LPS、球形红杆菌脂多糖（一种竞争性LPS拮抗剂）或紫杉醇（一种LPS激动剂）拮抗，并且它们对多粘菌素B处理具有相对抗性。然而，口腔细菌LPS对D-半乳糖胺处理的C3H/HeN小鼠的毒性低于沙门氏菌的LPS。这些发现表明，牙龈卟啉单胞菌和中间普氏菌的LPS的活性分子和作用方式与沙门氏菌的LPS有很大不同。
