Fluorescence image analysis of the association between Porphyromonas gingivalis and gingival epithelial cells

被引:105
作者
Belton, CM
Izutsu, KT [1 ]
Goodwin, PC
Park, Y
Lamont, RJ
机构
[1] Univ Washington, Dept Oral Biol, Seattle, WA 98195 USA
[2] Fred Hutchinson Canc Res Ctr, Seattle, WA 98109 USA
关键词
D O I
10.1046/j.1462-5822.1999.00022.x
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
We have developed a fluorescence imaging technique using a DNA-binding dye to visualize, over time, the physical interactions between Porphyromonas gingivalis and human gingival epithelial cells in vitro. The results extend previous observations of P. gingivalis invasion of gingival epithelial cells based on indirect measurements. An intracellular location for P. gingivalis was established by optical sectioning of images in the z-plane. Kinetic analysis showed that P. gingivalis invasion of epithelial cells is a rapid and efficient process, reaching completion after 12 min. Imaging of infected monolayers revealed that over 90% of a population of gingival epithelial cells contained bacteria. Furthermore, only vital bacteria were capable of invasion, and intracellular bacteria congregated in the perinuclear region of the epithelial cells. P. gingivalis remained inside the epithelial cells over a 24 h period and induced rearrangement of the actin cytoskeleton along with alteration of the size and shape of the epithelial cells. These findings provide direct evidence that entry rates of P. gingivalis into gingival epithelial cells are high and rapid, and that internalized bacteria initially localize in a specific region of the epithelial cells.
引用
收藏
页码:215 / 223
页数:9
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