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Porphyromonas gingivalis FimA Fimbriae: Fimbrial Assembly by fimA Alone in the fim Gene Cluster and Differential Antigenicity among fimA Genotypes

机译:牙龈卟啉单FIma菌毛:菌毛大会通过FIma独自一人在FIm基因簇和抗原差基因型FIma中

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摘要

The periodontal pathogen Porphyromonas gingivalis colonizes largely through FimA fimbriae, composed of polymerized FimA encoded by fimA. fimA exists as a single copy within the fim gene cluster (fim cluster), which consists of seven genes: fimX, pgmA and fimA-E. Using an expression vector, fimA alone was inserted into a mutant from which the whole fim cluster was deleted, and the resultant complement exhibited a fimbrial structure. Thus, the genes of the fim cluster other than fimA were not essential for the assembly of FimA fimbriae, although they were reported to influence FimA protein expression. It is known that there are various genotypes for fimA, and it was indicated that the genotype was related to the morphological features of FimA fimbriae, especially the length, and to the pathogenicity of the bacterium. We next complemented the fim cluster-deletion mutant with fimA genes cloned from P. gingivalis strains including genotypes I to V. All genotypes showed a long fimbrial structure, indicating that FimA itself had nothing to do with regulation of the fimbrial length. In FimA fimbriae purified from the complemented strains, types I, II, and III showed slightly higher thermostability than types IV and V. Antisera of mice immunized with each purified fimbria principally recognized the polymeric, structural conformation of the fimbriae, and showed low cross-reactivity among genotypes, indicating that FimA fimbriae of each genotype were antigenically different. Additionally, the activity of a macrophage cell line stimulated with the purified fimbriae was much lower than that induced by Escherichia coli lipopolysaccharide.
机译:牙周病原体牙龈卟啉单胞菌主要通过由FimA编码的聚合FimA组成的FimA菌毛大量繁殖。 fimA作为单拷贝存在于fim基因簇(fim簇)中,该簇由七个基因组成:fimX,pgmA和fimA-E。使用表达载体,将单独的fimA插入到缺失了整个fim簇的突变体中,并且得到的补体显示出纤维结构。因此,尽管有报道称fimA簇中除fimA以外的其他基因对于FimA菌毛的组装不是必需的,尽管它们影响FimA蛋白的表达。已知fimA存在多种基因型,并且表明该基因型与FimA菌毛的形态特征,特别是长度和细菌的致病性有关。接下来,我们用从牙龈卟啉单胞菌菌株克隆的fimA基因(包括基因型I至V)对fim簇缺失突变体进行了补充。所有基因型均显示出长的纤维结构,表明FimA本身与纤维长度的调节无关。从互补菌株纯化的FimA菌毛中,I,II和III型的热稳定性比IV和V型略高。用每种纯化的菌毛免疫的小鼠的抗血清主要识别该菌毛的聚合结构构象,并且交叉亲和力低。基因型之间的反应性,表明每种基因型的FimA菌毛在抗原上都不同。此外,纯化的菌毛刺激的巨噬细胞系的活性远低于大肠杆菌的脂多糖诱导的。

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