Adherence of Aggregatibacter actinomycetemcomitans via serotype-specific polysaccharide antigens in lipopolysaccharides
Adherence of Aggregatibacter actinomycetemcomitans via serotype-specific polysaccharide antigens in lipopolysaccharides
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DOI：
10.1111/j.1399-302x.2007.00416.x
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发表时间：
2008-06-01
影响因子：
--
通讯作者：
Chen, C.
中科院分区：
文献类型：
--
作者：
Fujise, O.;Wang, Y.;Chen, C.
前言：革兰氏阴性放线菌是一种重要的牙周病原菌。这种细菌的一个显着特征是它能够在非生物表面形成坚韧的生物膜。菌毛和非菌毛粘附素都被认为是导致这种能力的原因。本研究利用Cre-loxP重组系统构建了细胞表面多糖生物合成基因的无标记突变体，用于鉴定非菌毛粘附素(S)。方法：以非菌毛放线菌伴生菌株ATCC29523(血清型a型)为模板，构建内毒素中特异性多糖抗原(SPA-a)缺失突变体。经多粘菌素B柱分离纯化，经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法银染和兔抗SPA-a抗血清免疫印迹分析。结果：ATCC29523菌株在玻璃表面形成了一层细菌生长的薄膜。SPA-a的缺失影响了其形成该薄膜的能力。当该突变体被野生型SPA-a基因簇抢救后，其黏附阳性表型得以恢复。结论：内毒素分子中的SpA-A可促进放线菌伴生放线菌细胞对非生物表面的黏附。
Introduction: Gram-negative Aggregatibacter actinomycetemcomitans is recognized as an important periodontal pathogen. A striking property of this bacterium is its ability to form a tenacious biofilm adhering to abiotic surfaces. Both fimbrial and non-fimbrial adhesins are believed to be responsible for this ability. In our study, specific markerless mutants in the biosynthesis genes of cell surface polysaccharides were constructed with the Cre-loxP recombination system to identify non-fimbrial adhesin(s).Methods: Non-fimbriated A. actinomycetemcomitans strain ATCC29523 (serotype a) was used to construct a deletion mutant of serotype-a specific polysaccharide antigen (SPA-a) in lipopolysaccharide (LPS). The LPS was purified through a polymyxin B column following phenol extraction, and verified by silver staining following sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by immunoblot analysis using rabbit antisera raised against SPA-a. Strains were grown in broth for 2 days and examined for the adherence of bacterial cells on the glass surface.Results: Strain ATCC29523 formed a thin film of bacterial growth on the glass surface. The deletion of SPA-a affected its ability to form this thin film. When this mutant was rescued with the wild-type SPA-a gene cluster, its adherence-positive phenotype was restored.Conclusion: SPA-a in the LPS molecule appears to promote the adherence of A. actinomycetemcomitans cells to abiotic surfaces.