Mintz K P, Fives-Taylor P M
Department of Microbiology and Molecular Genetics, Markey Center for Molecular Genetics, College of Medicine, Burlington, Vermont.
Infect Immun. 1994 Sep;62(9):3672-8. doi: 10.1128/iai.62.9.3672-3678.1994.
Two quantitative, rapid assays were developed to study the adhesion of Actinobacillus actinomycetemcomitans, an oral bacterium associated with periodontal disease, to human epithelial cells. The human oral carcinoma cell line KB was grown in microtiter plates, and adherent bacteria were detected by an enzyme-linked immunosorbent assay with purified anti-A. actinomycetemcomitans serum and horseradish peroxidase-conjugated secondary antibody or [3H]thymidine-labeled bacteria. Adhesion was found to be time dependent and increased linearly with increasing numbers of bacteria added. Variation in the level of adhesion was noted among strains of A. actinomycetemcomitans. Adhesion was not significantly altered by changes in pH (from pH 5 to 9) but was sensitive to sodium chloride concentrations greater than 0.15 M. Pooled human saliva was inhibitory for adhesion when bacteria were pretreated with saliva before being added to the cells. Pretreatment of the KB cells with saliva did not inhibit adhesion. Protease treatment of A. actinomycetemcomitans reduced adhesion of the bacteria to KB cells. The data are consistent with the hypothesis that a protein(s) is required for bacterial adhesion and that host components may play a role in modulating adhesion to epithelial cells.
开发了两种定量快速检测方法,用于研究伴放线放线杆菌(一种与牙周疾病相关的口腔细菌)对人上皮细胞的黏附。人口腔癌细胞系KB在微量滴定板中培养,通过酶联免疫吸附测定法,使用纯化的抗伴放线放线杆菌血清和辣根过氧化物酶偶联的二抗或[3H]胸苷标记的细菌来检测黏附的细菌。发现黏附具有时间依赖性,并且随着添加细菌数量的增加呈线性增加。观察到伴放线放线杆菌不同菌株之间黏附水平存在差异。pH值从5到9的变化对黏附没有显著影响,但对大于0.15 M的氯化钠浓度敏感。当细菌在添加到细胞之前用唾液预处理时,混合的人唾液对黏附具有抑制作用。用唾液预处理KB细胞不会抑制黏附。用蛋白酶处理伴放线放线杆菌会降低细菌对KB细胞的黏附。这些数据与以下假设一致:细菌黏附需要一种或多种蛋白质,并且宿主成分可能在调节对上皮细胞的黏附中起作用。