Saunders J M, Miller C H
Infect Immun. 1980 Sep;29(3):981-9. doi: 10.1128/iai.29.3.981-989.1980.
A standardized assay was used to measure the attachment of Actinomyces naeslundii ATCC 12104 to washed human buccal epithelial cells. Treatment of the A. naeslundii cells with hyaluronidases, wheat germ lipase, protease, trypsin, heat, or sonic oscillation significantly reduced their ability to attach to epithelial cells. Treatment of the epithelial cells with the above enzymes did not influence the attachment of A. naeslundii. Extraction of A. naeslundii with NaOH also significantly reduced the ability of the bacterium to attach to human buccal epithelial cells. The neutralized and dialyzed NaOH extract contained both carbohydrate and protein substances in a ratio of about 1:1. Adding this extract back to the extracted bacterial cells partially restored their ability to attach to epithelial cells. When the NaOH extract was preincubated with epithelial cells and residual extract was removed by washing, attachment of normal A. naeslundii was partially blocked. The ability of the extracted material to block attachment was significantly reduced when treated with hyaluronidases or with wheat germ lipase. Treatment with heat, protease, or trypsin did not significantly reduce the ability of the extracted materials to block attachment. Pretreatment of the epithelial cells with hyaluronic acid or chondroitin sulfate also reduced subsequent attachment of normal A. naeslundii cells. Pretreatment of epithelial cells with dextrans, proteins, or unpure mannose did not influence subsequent attachment of A. naeslundii. Pretreatment of A. naeslundii with galactose and lactose significantly inhibited attachment to normal epithelial cells. The results suggest that the attachment of A. naeslundii to human buccal epithelial cells may involve mucopolysaccharides similar to hyaluronic acid on the surface of the bacterial cells. Other attachment mechanisms may also be operative.
采用标准化检测方法来测定内氏放线菌ATCC 12104对洗涤过的人颊上皮细胞的黏附情况。用透明质酸酶、小麦胚芽脂肪酶、蛋白酶、胰蛋白酶、加热或超声振荡处理内氏放线菌细胞,可显著降低其黏附上皮细胞的能力。用上述酶处理上皮细胞,并不影响内氏放线菌的黏附。用氢氧化钠提取内氏放线菌,也显著降低了该菌黏附人颊上皮细胞的能力。经中和及透析的氢氧化钠提取物含有碳水化合物和蛋白质物质,二者比例约为1:1。将该提取物重新添加到提取后的细菌细胞中,可部分恢复其黏附上皮细胞的能力。当氢氧化钠提取物与上皮细胞预孵育,然后通过洗涤去除残留提取物时,正常内氏放线菌的黏附会受到部分阻断。用透明质酸酶或小麦胚芽脂肪酶处理后,提取物阻断黏附的能力显著降低。用加热、蛋白酶或胰蛋白酶处理,不会显著降低提取物阻断黏附的能力。用透明质酸或硫酸软骨素预处理上皮细胞,也会降低正常内氏放线菌细胞随后的黏附。用葡聚糖、蛋白质或不纯的甘露糖预处理上皮细胞,不影响内氏放线菌随后的黏附。用半乳糖和乳糖预处理内氏放线菌,可显著抑制其对正常上皮细胞的黏附。结果表明,内氏放线菌与人颊上皮细胞的黏附可能涉及细菌细胞表面类似于透明质酸的黏多糖。其他黏附机制可能也起作用。