Rapid paracellular transmigration of Campylobacter jejuni across polarized epithelial cells without affecting TER: role of proteolytic-active HtrA cleaving E-cadherin but not fibronectin

被引:113
作者
Boehm, Manja [1 ]
Hoy, Benjamin [3 ]
Rohde, Manfred [4 ]
Tegtmeyer, Nicole [1 ]
Baek, Kristoffer T. [5 ]
Oyarzabal, Omar A. [6 ]
Brondsted, Lone [5 ]
Wessler, Silja [3 ]
Backert, Steffen [1 ,2 ]
机构
[1] Univ Coll Dublin, Sch Med & Med Sci, Dublin 4, Ireland
[2] Univ Coll Dublin, UCD Sch Biomol & Biomed Sci, Dublin 4, Ireland
[3] Salzburg Univ, Div Microbiol, A-5020 Salzburg, Austria
[4] Helmholtz Ctr Infect Res, Dept Med Microbiol, D-38124 Braunschweig, Germany
[5] Univ Copenhagen, Dept Vet Dis Biol, DK-1870 Frederiksberg C, Denmark
[6] Inst Environm Hlth Inc, Seattle, WA 98155 USA
关键词
HtrA; E-cadherin; Fibronectin; MKN-28; Molecular pathogenesis; Cellular invasion; Signaling; TER; Virulence; TRANSCELLULAR TRANSLOCATION; BACTERIAL INVASION; TIGHT JUNCTIONS; CACO-2; CELLS; PROTEIN; ADHERENCE; VIRULENCE; PROTEASES; BINDING; GENE;
D O I
10.1186/1757-4749-4-3
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
Background: Campylobacter jejuni is one of the most important bacterial pathogens causing food-borne illness worldwide. Crossing the intestinal epithelial barrier and host cell entry by C. jejuni 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. Results: In the present study, we characterized the serine protease HtrA (high-temperature requirement A) of C. jejuni as a secreted virulence factor with important proteolytic functions. Infection studies and in vitro cleavage assays showed that C. jejuni'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 H. pylori's HtrA. Deletion of the htrA gene in C. jejuni 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 C. jejuni is highly efficient and is achieved within a few minutes of infection. Interestingly, E-cadherin cleavage by C. jejuni 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 Salmonella, Shigella, Listeria or Neisseria. Conclusion: These results suggest that HtrA-mediated E-cadherin cleavage is involved in rapid crossing of the epithelial barrier by C. jejuni via a very specific mechanismusing the paracellular route to reach basolateral surfaces, but does not cleave the fibronectin receptor which is necessary for cell entry.
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