Breeling J L, Onderdonk A B, Cisneros R L, Kasper D L
Division of Infectious Diseases, Brigham and Women's Hospital, Boston, Massachusetts 02115.
Infect Immun. 1988 Jul;56(7):1754-9. doi: 10.1128/iai.56.7.1754-1759.1988.
Previous experiments with the carrageenan model for ulcerative colitis demonstrated that the inflammatory response in guinea pigs can be enhanced by immunization with Bacteroides vulgatus and subsequent feeding of this organism to experimental animals. The studies reported here show that antigens extractable from the bacterial outer membrane by EDTA are responsible for this effect. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to analyze the outer membrane proteins from various strains as well as the lipopolysaccharides (LPS) extractable by the phenol-water method. Although the observed pattern of outer membrane proteins was complex, the strains could be divided into two electrophoretic types (phenons) on the basis of immunoblotting against a panel of antisera. Cross-absorbed antisera used in immunoblotting experiments identified four outer membrane proteins uniquely associated with the phenon capable of enhancing the colitis inflammatory response. These proteins had molecular weights of 100,000, 57,000, 34,000, and 21,000 when measured in 8% to 12% acrylamide gradient sodium dodecyl sulfate gels. Other antigens identified included at least one type of smooth LPS, three types of rough LPS, and a common antigen of 30,000 molecular weight among the strains of B. vulgatus tested. The outer membrane preparations were used in animal immunization and challenge experiments, and the severity of colitis was correlated with one electrophoresis type. The potential role of membrane proteins in the enhancement of colitis is discussed.
先前用角叉菜胶诱导溃疡性结肠炎模型进行的实验表明,用普通拟杆菌免疫豚鼠,随后将该菌投喂给实验动物,可增强其炎症反应。本文报道的研究表明,通过乙二胺四乙酸(EDTA)从细菌外膜提取的抗原是造成这种效应的原因。采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析了不同菌株的外膜蛋白以及用酚水法可提取的脂多糖(LPS)。尽管观察到的外膜蛋白图谱很复杂,但根据针对一组抗血清的免疫印迹分析,这些菌株可分为两种电泳类型(菌型)。免疫印迹实验中使用的交叉吸收抗血清鉴定出四种外膜蛋白,它们与能够增强结肠炎炎症反应的菌型独特相关。在8%至12%丙烯酰胺梯度十二烷基硫酸钠凝胶中测定时,这些蛋白的分子量分别为100,000、57,000、34,000和21,000。鉴定出的其他抗原包括至少一种光滑型LPS、三种粗糙型LPS以及受试普通拟杆菌菌株中一种分子量为30,000的共同抗原。将外膜制剂用于动物免疫和激发实验,结肠炎的严重程度与一种电泳类型相关。文中讨论了膜蛋白在增强结肠炎方面的潜在作用。