McArthur W P, Stroup S, McClellan S, Leung K P
Department of Oral Biology, College of Dentistry, University of Florida, Gainesville, USA.
Oral Microbiol Immunol. 1996 Aug;11(4):209-19. doi: 10.1111/j.1399-302x.1996.tb00172.x.
The serotype b antigens have been reported to be associated with lipopolysaccharide. Using murine monoclonal antibodies specific for either a serotype b antigen or the Actinobacillus actinomycetemcomitans species, the relationship of the two epitopes to lipopolysaccharide was determined. Both the species-specific and serotype b-specific monoclonal antibodies bound to whole cells, vesicles and conventionally isolated lipopolysaccharide and polysaccharide material derived from A. actinomycetemcomitans culture supernatants. Serotype b-specific monoclonal antibodies bound to the polysaccharide of acid-hydrolyzed lipopolysaccharide. Species-specific monoclonal antibodies bound to both the polysaccharide and the lipid A fraction of lipopolysaccharide after acid hydrolysis. Polymyxin b partially inhibited the binding of the species-specific monoclonal antibodies to lipopolysaccharide and had no effect on the binding of the serotype b-specific monoclonal antibodies to lipopolysaccharide. Lipopolysaccharide from whole bacteria and polysaccharide material isolated from culture supernatants were separated by gel filtration chromatography in deoxycholate into fractions that contained serotype b antigen, both serotype b and species-specific antigens, or species-specific antigen. SDS-polyacrylamide gel electrophoresis and Western blotting analysis of the fractions revealed that the serotype b antigen was on a high-molecular-weight polysaccharide material. The species-specific antigen was on a ladder of lower-molecular-weight polysaccharides identical to the blot pattern of lipopolysaccharide molecules separated by polyacrylamide gel electrophoresis and stained with silver stain. Chemical analysis of the polysaccharide containing serotype b antigen revealed 85% ribose, 11% glucose, and no lipid. Chemical content of the species-specific antigenic material revealed a composition typical of lipopolysaccharide. Immunoelectron microscopy using the species- or serotype b-specific monoclonal antibodies confirmed the biochemical and immunological characterization of the two antigens, showing that the species-specific epitopes were on the surface of the A. actinomycetemcomitans cell membrane and the serotype b-specific epitopes on the amorphous material extending from the cell surface. The data indicated that the serotype b antigen, detected by the antibody, was separable from lipopolysaccharide and was an A. actinomycetemcomitans capsular material. The species-specific antigen, being more conserved than the serotype antigen, was on all the lipopolysaccharide molecular species.
据报道，b 型血清型抗原与脂多糖有关。使用针对 b 型血清型抗原或伴放线放线杆菌的鼠单克隆抗体，确定了这两种表位与脂多糖的关系。种特异性和 b 型血清型特异性单克隆抗体均与全细胞、囊泡以及常规分离的脂多糖和源自伴放线放线杆菌培养上清液的多糖物质结合。b 型血清型特异性单克隆抗体与酸水解脂多糖的多糖结合。种特异性单克隆抗体在酸水解后与脂多糖的多糖和脂质 A 部分结合。多粘菌素 b 部分抑制种特异性单克隆抗体与脂多糖的结合，对 b 型血清型特异性单克隆抗体与脂多糖的结合没有影响。来自全菌的脂多糖和从培养上清液中分离的多糖物质通过在脱氧胆酸盐中的凝胶过滤色谱法分离成含有 b 型血清型抗原、b 型血清型和种特异性抗原或种特异性抗原的级分。对这些级分的 SDS-聚丙烯酰胺凝胶电泳和 Western 印迹分析表明，b 型血清型抗原存在于高分子量多糖物质上。种特异性抗原存在于一系列低分子量多糖上，与通过聚丙烯酰胺凝胶电泳分离并用银染染色的脂多糖分子的印迹模式相同。对含有 b 型血清型抗原的多糖的化学分析显示含有 85%的核糖、11%的葡萄糖，且无脂质。种特异性抗原物质的化学组成显示出典型的脂多糖组成。使用种特异性或 b 型血清型特异性单克隆抗体的免疫电子显微镜证实了这两种抗原的生化和免疫学特征，表明种特异性表位存在于伴放线放线杆菌细胞膜表面，b 型血清型特异性表位存在于从细胞表面延伸的无定形物质上。数据表明，抗体检测到的 b 型血清型抗原可与脂多糖分离，是伴放线放线杆菌的荚膜物质。种特异性抗原比血清型抗原更保守，存在于所有脂多糖分子种类上。
