Rapid paracellular transmigration of <it>Campylobacter jejuni</it> across polarized epithelial cells without affecting TER: role of proteolytic-active HtrA cleaving E-cadherin but not fibronectin
<p>Abstract</p> <p>Background</p> <p><it>Campylobacter jejuni</it> is one of the most important bacterial pathogens causing food-borne illness worldwide. Crossing the intestinal epithelial barrier and host cell entry by <it>C. jejuni</it> is cons...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2012-04-01
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| Series: | Gut Pathogens |
| Subjects: | |
| Online Access: | http://www.gutpathogens.com/content/4/1/3 |
| _version_ | 1819122195360645120 |
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| author | Boehm Manja Hoy Benjamin Rohde Manfred Tegtmeyer Nicole Bæk Kristoffer T Oyarzabal Omar A Brøndsted Lone Wessler Silja Backert Steffen |
| author_facet | Boehm Manja Hoy Benjamin Rohde Manfred Tegtmeyer Nicole Bæk Kristoffer T Oyarzabal Omar A Brøndsted Lone Wessler Silja Backert Steffen |
| author_sort | Boehm Manja |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p><it>Campylobacter jejuni</it> is one of the most important bacterial pathogens causing food-borne illness worldwide. Crossing the intestinal epithelial barrier and host cell entry by <it>C. jejuni</it> is considered the primary reason of damage to the intestinal tissue, but the molecular mechanisms as well as major bacterial and host cell factors involved in this process are still widely unclear.</p> <p>Results</p> <p>In the present study, we characterized the serine protease HtrA (high-temperature requirement A) of <it>C. jejuni</it> as a secreted virulence factor with important proteolytic functions. Infection studies and in vitro cleavage assays showed that <it>C. jejuni</it>’s HtrA triggers shedding of the extracellular E-cadherin NTF domain (90 kDa) of non-polarised INT-407 and polarized MKN-28 epithelial cells, but fibronectin was not cleaved as seen for <it>H. pylori</it>’s HtrA. Deletion of the <it>htrA</it> gene in <it>C. jejuni</it> or expression of a protease-deficient S197A point mutant did not lead to loss of flagella or reduced bacterial motility, but led to severe defects in E-cadherin cleavage and transmigration of the bacteria across polarized MKN-28 cell layers. Unlike other highly invasive pathogens, transmigration across polarized cells by wild-type <it>C. jejuni</it> is highly efficient and is achieved within a few minutes of infection. Interestingly, E-cadherin cleavage by <it>C. jejuni</it> occurs in a limited fashion and transmigration required the intact flagella as well as HtrA protease activity, but does not reduce transepithelial electrical resistance (TER) as seen with <it>Salmonella, Shigella</it>, <it>Listeria</it> or <it>Neisseria.</it></p> <p>Conclusion</p> <p>These results suggest that HtrA-mediated E-cadherin cleavage is involved in rapid crossing of the epithelial barrier by <it>C. jejuni</it> via a very specific mechanism using the paracellular route to reach basolateral surfaces, but does not cleave the fibronectin receptor which is necessary for cell entry.</p> |
| first_indexed | 2024-12-22T06:48:36Z |
| format | Article |
| id | doaj.art-7ebce51b7e5948d58158d03e5a7b34a4 |
| institution | Directory Open Access Journal |
| issn | 1757-4749 |
| language | English |
| last_indexed | 2024-12-22T06:48:36Z |
| publishDate | 2012-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Gut Pathogens |
| spelling | doaj.art-7ebce51b7e5948d58158d03e5a7b34a42022-12-21T18:35:13ZengBMCGut Pathogens1757-47492012-04-0141310.1186/1757-4749-4-3Rapid paracellular transmigration of <it>Campylobacter jejuni</it> across polarized epithelial cells without affecting TER: role of proteolytic-active HtrA cleaving E-cadherin but not fibronectinBoehm ManjaHoy BenjaminRohde ManfredTegtmeyer NicoleBæk Kristoffer TOyarzabal Omar ABrøndsted LoneWessler SiljaBackert Steffen<p>Abstract</p> <p>Background</p> <p><it>Campylobacter jejuni</it> is one of the most important bacterial pathogens causing food-borne illness worldwide. Crossing the intestinal epithelial barrier and host cell entry by <it>C. jejuni</it> is considered the primary reason of damage to the intestinal tissue, but the molecular mechanisms as well as major bacterial and host cell factors involved in this process are still widely unclear.</p> <p>Results</p> <p>In the present study, we characterized the serine protease HtrA (high-temperature requirement A) of <it>C. jejuni</it> as a secreted virulence factor with important proteolytic functions. Infection studies and in vitro cleavage assays showed that <it>C. jejuni</it>’s HtrA triggers shedding of the extracellular E-cadherin NTF domain (90 kDa) of non-polarised INT-407 and polarized MKN-28 epithelial cells, but fibronectin was not cleaved as seen for <it>H. pylori</it>’s HtrA. Deletion of the <it>htrA</it> gene in <it>C. jejuni</it> or expression of a protease-deficient S197A point mutant did not lead to loss of flagella or reduced bacterial motility, but led to severe defects in E-cadherin cleavage and transmigration of the bacteria across polarized MKN-28 cell layers. Unlike other highly invasive pathogens, transmigration across polarized cells by wild-type <it>C. jejuni</it> is highly efficient and is achieved within a few minutes of infection. Interestingly, E-cadherin cleavage by <it>C. jejuni</it> occurs in a limited fashion and transmigration required the intact flagella as well as HtrA protease activity, but does not reduce transepithelial electrical resistance (TER) as seen with <it>Salmonella, Shigella</it>, <it>Listeria</it> or <it>Neisseria.</it></p> <p>Conclusion</p> <p>These results suggest that HtrA-mediated E-cadherin cleavage is involved in rapid crossing of the epithelial barrier by <it>C. jejuni</it> via a very specific mechanism using the paracellular route to reach basolateral surfaces, but does not cleave the fibronectin receptor which is necessary for cell entry.</p>http://www.gutpathogens.com/content/4/1/3HtrAE-cadherinFibronectinMKN-28Molecular pathogenesisCellular invasionSignalingTERVirulence |
| spellingShingle | Boehm Manja Hoy Benjamin Rohde Manfred Tegtmeyer Nicole Bæk Kristoffer T Oyarzabal Omar A Brøndsted Lone Wessler Silja Backert Steffen Rapid paracellular transmigration of <it>Campylobacter jejuni</it> across polarized epithelial cells without affecting TER: role of proteolytic-active HtrA cleaving E-cadherin but not fibronectin Gut Pathogens HtrA E-cadherin Fibronectin MKN-28 Molecular pathogenesis Cellular invasion Signaling TER Virulence |
| title | Rapid paracellular transmigration of <it>Campylobacter jejuni</it> across polarized epithelial cells without affecting TER: role of proteolytic-active HtrA cleaving E-cadherin but not fibronectin |
| title_full | Rapid paracellular transmigration of <it>Campylobacter jejuni</it> across polarized epithelial cells without affecting TER: role of proteolytic-active HtrA cleaving E-cadherin but not fibronectin |
| title_fullStr | Rapid paracellular transmigration of <it>Campylobacter jejuni</it> across polarized epithelial cells without affecting TER: role of proteolytic-active HtrA cleaving E-cadherin but not fibronectin |
| title_full_unstemmed | Rapid paracellular transmigration of <it>Campylobacter jejuni</it> across polarized epithelial cells without affecting TER: role of proteolytic-active HtrA cleaving E-cadherin but not fibronectin |
| title_short | Rapid paracellular transmigration of <it>Campylobacter jejuni</it> across polarized epithelial cells without affecting TER: role of proteolytic-active HtrA cleaving E-cadherin but not fibronectin |
| title_sort | rapid paracellular transmigration of it campylobacter jejuni it across polarized epithelial cells without affecting ter role of proteolytic active htra cleaving e cadherin but not fibronectin |
| topic | HtrA E-cadherin Fibronectin MKN-28 Molecular pathogenesis Cellular invasion Signaling TER Virulence |
| url | http://www.gutpathogens.com/content/4/1/3 |
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