Journal List > J Bacteriol Virol > v.36(3) > 1033838

Kang, Piao, Shin, Lee, and Oh: Adhesion of Weissella cibaria to the Epithelial Cells and Factors Affecting its Adhesion

Abstract

We evaluated the ability of lactic acid bacteria, Weissella cibaria, isolated from the oral cavity to adhere to epithelial cells. W. cibaria efficiently adhered to KB cells and HeLa cells. In addition, W. cibaria efficiently adhered to Fusobacterium nucleatum. But the adhesiveness of W. cibaria disappeared upon exposure to LiCl or pronase, suggesting that the S-layer proteins of W. cibaria mediated the adhesiveness. The molecular mass of the S-layer proteins extracted from W. cibaria was approximately 50 kDa. When W. cibaria strains were washed with 0.45% saline, the bacteria were efficiently adhered to the epithelial cells. In conclusion, W. cibaria has the ability to adhere to epithelial cells through the S-layer proteins.

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Figure 1.
Effect of 5 M LiCl treatment on the adherence of W. cibaria CMS2 to epithelial cells. A, untreated W. cibaria CMS2 vs. KB cells; B, treated W. cibaria CMS2 vs. KB cells; C, untreated W. cibaria CMS2 vs. HeLa cells; D, treated W. cibaria CMS2 vs. HeLa cells. Magnification, × 1000. Shown in the figures are the representative data of three separate experiments.
jbv-36-151f1.tif
Figure 2.
Numbers of adhered W. cibaria CMS2 to epithelial cells. Treatment of W. cibaria CMS2 was as follows: ☐, control; ▪, 5 M LiCl. Values are expressed as means ± SD. ∗∗Significances of differences were defined as p<0.05.
jbv-36-151f2.tif
Figure 3.
SDS-PAGE of S-layer proteins of W. cibaria strains extracted by LiCl (5 mol l–1) treatment. Lane 1, W. cibaria CMU; lane 2, W. cibaria CMS2; lane 3, W. cibaria CMS3. Shown in the figures are the representative data of three separate experiments.
jbv-36-151f3.tif
Figure 4.
Microscopic observations of adhesion between W. cibaria strain and F. nucleatum ATCC 10953. A, bright-field micrographs of W. cibaria CMS2 vs. F. nucleatum; Magnification, × 1000; B, TEM micrographs of untreated W. cibaria CMS2 whole cell only; C, and D, TEM micrographs of adhesion between W. cibaria CMS2 and F. nucleatum before and after pronase treatment, respectively. Wc represents W. cibaria; Fn represents F. nucleatum. Shown in the figures are the representative data of three separate experiments.
jbv-36-151f4.tif
Figure 5.
Effect of incubation temperature and saline solution on the adherence of W. cibaria CMS2 to KB cells. A, bacterial cells were incubated at 30°C and washed with 0.9% saline; B, bacterial cells were incubated at 30°C and washed with 0.45% saline; C, bacterial cells were incubated at 30°C and washed with distilled water; D, bacterial cells were incubated at 37°C and washed with 0.9% saline; E, bacterial cells were incubated at 37°C and washed with 0.45% saline; F, bacterial cells were incubated at 37°C and washed with distilled water. Magnification, × 1000. Shown in the figures are the representative data of three separate experiments.
jbv-36-151f5.tif
Figure 6.
Effect of incubation temperature and saline solution on the adherence of W. cibaria CMS2 to HeLa cells. A, bacterial cells were incubated at 30°C and washed with 0.9% saline; B, bacterial cells were incubated at 30°C and washed with 0.45% saline; C, bacterial cells were incubated at 30°C and washed with distilled water; D, bacterial cells were incubated at 37°C and washed with 0.9% saline; E, bacterial cells were incubated at 37°C and washed with 0.45% saline; F, bacterial cells were incubated at 37°C and washed with distilled water. Magnification, × 1000. Shown in the figures are the representative data of three separate experiments.
jbv-36-151f6.tif
Figure 7.
Numbers of adhered W. cibaria CMS2 to epithelial cells at various conditions. A, adherence of W. cibaria CMS2 to KB cell; B, adherence of W. cibaria CMS2 to HeLa cell. W. cibaria CMS2 were incubated at 30°C or 37°C and washed with saline solutions as follows: ▪, 0.9% saline; ☐, 0.45% saline; ▪, distilled water. Means ± SD followed by distinct letters differ statistically (p<0.05).
jbv-36-151f7.tif
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