<i>Treponema denticola</i> Induces Epithelial Barrier Dysfunction in Polarized Epithelial Cells
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- Kikuchi Yuichiro
- Department of Microbiology, Tokyo Dental College Oral Health Science Center, Tokyo Dental College
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- Kimizuka Ryuta
- Department of Microbiology, Tokyo Dental College
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- Kato Tetsuo
- Laboratory of Chemistry, Tokyo Dental College
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- Okuda Katsuji
- Department of Microbiology, Tokyo Dental College
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- Kokubu Eitoyo
- Department of Microbiology, Tokyo Dental College Oral Health Science Center, Tokyo Dental College
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- Ishihara Kazuyuki
- Department of Microbiology, Tokyo Dental College Oral Health Science Center, Tokyo Dental College
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Abstract
<p>Treponema denticola, an anaerobic spirochete found mainly in the oral cavity, is associated with periodontal disease and has a variety of virulence factors. Although in vitro studies have shown that T. denticola is able to penetrate epithelial cell monolayers, its effect on the epithelial barrier junction is not known. Human gingival epithelial cells are closely associated with adjacent membranes, forming barriers in the presence of tight junction proteins, including zonula occludens-1 (ZO-1), claudin-1, and occludin. Tight junction proteins are also expressed by Madin-Darby canine kidney (MDCK) cells in culture. In this study, the MDCK cell profile was investigated following infection with T. denticola (ATCC 35405) wild-type, as well as with its dentilisin-deficient mutant, K1. Basolateral exposure of MDCK cell monolayers to T. denticola at a multiplicity of infection (MOI) of 104 resulted in a decrease in transepithelial electrical resistance (TER). Transepithelial electrical resistance in MDCK cell monolayers also decreased following apical exposure to T. denticola (MOI=104), although this took longer with basolateral exposure. The effect on the TER was time-dependent and required the presence of live bacteria. Meanwhile, MDCK cell viability showed a decrease with either basolateral or apical exposure. Immunofluorescence analysis demonstrated decreases in the amounts of immunoreactive ZO-1 and claudin-1 in association with disruption of cell-cell junctions in MDCK cells exposed apically or basolaterally to T. denticola. Western blot analysis demonstrated degradation of ZO-1 and claudin-1 in culture lysates derived from T. denticola-exposed MDCK cells, suggesting a bacteria-induced protease capable of cleaving these tight junction proteins.</p>
Journal
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- The Bulletin of Tokyo Dental College
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The Bulletin of Tokyo Dental College 59 (4), 265-275, 2018
Tokyo Dental College, Japan