Characterization of <i>Porphyromonas gingivalis</i> -Induced Degradation of Epithelial Cell Junctional Complexes

Infection and Immunity - Tập 68 Số 3 - Trang 1441-1449 - 2000
Jannet Katz1, Vijaya Sambandam2,3, John H. Wu2, Suzanne M. Michalek4,1, Daniel F. Balkovetz5,2,3
1Department of Oral Biology,1
2Department of Medicine 2 and
3Veterans Administration Medical Center, Birmingham, Alabama 352333
4Department of Microbiology,4
5Department of Cell Biology,5 University of Alabama at Birmingham, Birmingham, Alabama 35294, and

Tóm tắt

ABSTRACT Porphyromonas gingivalis is considered among the etiological agents of human adult periodontitis. Although in vitro studies have shown that P. gingivalis has the ability to invade epithelial cell lines, its effect on the epithelial barrier junctions is not known. Immunofluorescence analysis of human gingival epithelial cells confirmed the presence of tight-junction (occludin), adherens junction (E-cadherin), and cell-extracellular matrix junction (β1-integrin) transmembrane proteins. These transmembrane proteins are expressed in Madin-Darby canine kidney (MDCK) cells. In addition, MDCK cells polarize and therefore serve as a useful in vitro model for studies on the epithelial cell barrier. Using the MDCK cell system, we examined the effect of P. gingivalis on epithelial barrier function. Exposure of the basolateral surfaces of MDCK cells to P. gingivalis (>10 9 bacteria/ml) resulted in a decrease in transepithelial resistance. Immunofluorescence microscopy demonstrated decreases in the amounts of immunoreactive occludin, E-cadherin, and β1-integrin at specific times which were related to a disruption of cell-cell junctions in MDCK cells exposed to basolateral P. gingivalis . Disruption of cell-cell junctions was also observed upon apical exposure to bacteria; however, the effects took longer than those seen upon basolateral exposure. Cell viability was not affected by either basolateral or apical exposure to P. gingivalis . Western blot analysis demonstrated hydrolysis of occludin, E-cadherin, and β1-integrin in lysates derived from MDCK cells exposed to P. gingivalis . Immunoprecipitated occludin and E-cadherin molecules from MDCK cell lysates were also degraded by P. gingivalis , suggesting a bacterial protease(s) capable of cleaving these epithelial junction transmembrane proteins. Collectively, these data suggest that P. gingivalis is able to invade the deeper structures of connective tissues via a paracellular pathway by degrading epithelial cell-cell junction complexes, thus allowing the spread of the bacterium. These results also indicate the importance of a critical threshold concentration of P. gingivalis to initiate epithelial barrier destruction.

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Tài liệu tham khảo

10.1128/iai.63.12.4744-4754.1995

10.1016/S0024-3205(99)00073-9

10.1074/jbc.272.6.3471

10.1128/jb.178.10.2734-2741.1996

10.1902/jop.1996.67.10s.1123

10.1034/j.1399-302X.1999.140301.x

10.1016/S0021-9258(19)37045-0

10.1128/jb.176.15.4549-4557.1994

10.1099/00221287-139-5-949

10.1128/iai.61.5.2260-2265.1993

10.1128/iai.61.3.940-946.1993

10.1681/ASN.V1061337

10.1083/jcb.107.1.221

10.1128/iai.65.7.2861-2867.1997

10.1111/j.1399-302X.1990.tb00441.x

10.1083/jcb.123.6.1777

10.1083/jcb.102.2.457

10.1083/jcb.141.1.199

10.1111/j.1399-302X.1996.tb00202.x

10.1177/10454411910020020301

10.1177/00220345900690120601

Isberg R. R. Pathways for the penetration of enteroinvasive Yersinia into mammalian cells.Mol. Biol. Med.719907382

10.1016/0092-8674(90)90099-Z

10.1016/0003-9969(88)90028-3

10.1203/00006450-199712000-00014

10.1128/iai.60.9.3579-3585.1992

10.1111/j.1399-302X.1996.tb00187.x

10.1128/iai.63.10.3878-3885.1995

10.1111/j.1600-0765.1993.tb02104.x

10.1083/jcb.110.3.803

Larjava H. Zhou C. Larjava I. Rahemtulla F. Immunolocalization of β1 integrins in human gingival epithelium and cultured keratinocytes.Scand. J. Dent. Res.1001992266273

10.1016/S0092-8674(00)81070-3

10.1128/iai.64.8.2988-2997.1996

10.1128/iai.57.2.566-573.1989

10.1038/329341a0

10.1128/iai.65.4.1431-1439.1997

10.1902/jop.1996.67.10s.1103

10.1128/iai.57.11.3466-3471.1989

10.1093/oxfordjournals.jbchem.a021426

10.1128/iai.54.3.659-665.1986

10.1074/jbc.272.3.1595

10.1016/S0021-9258(17)42365-9

10.1128/iai.63.4.1176-1182.1995

10.1111/j.1399-302X.1989.tb00238.x

10.1111/j.1399-302X.1995.tb00160.x

10.1128/iai.60.9.3909-3912.1992

10.1126/science.2672330

10.1902/jop.1988.59.4.259

10.1111/j.1600-0765.1993.tb01072.x

10.1111/j.1600-0765.1994.tb01092.x

10.1242/jcs.107.2.527

10.1128/IAI.67.4.1837-1843.1999

10.1002/(SICI)1234-987X(199607)9:4<167::AID-MEH425>3.3.CO;2-I

10.1128/iai.61.4.1239-1245.1993

10.1111/j.1600-0765.1982.tb01137.x

10.1128/jcm.11.5.522-526.1980

10.1128/iai.21.3.821-829.1978

10.1016/0167-7012(93)90038-J

10.1128/IAI.66.3.1159-1166.1998

10.1177/10454411910020030301

10.1111/j.1600-0765.1982.tb01129.x

10.1172/JCI5844

10.1128/iai.65.1.313-316.1997

10.1016/0003-9969(90)90016-4

10.1073/pnas.95.25.14979

10.1111/j.1348-0421.1987.tb03154.x

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Infection and Immunity

Tập 68 Số 3

1441-1449

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