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Microbiol."],"abstract":"<jats:p><jats:italic>Cronobacter sakazakii<\/jats:italic> (<jats:italic>C. sakazakii<\/jats:italic>) is an emerging opportunistic foodborne pathogen that can cause neonatal necrotizing enterocolitis, meningitis, sepsis in neonates and infants with a relatively high mortality rate. Bacterial transcytosis across the human brain microvascular endothelial cells (HBMEC) is vital for <jats:italic>C. sakazakii<\/jats:italic> to induce neonatal meningitis. However, few studies focus on the mechanisms by which <jats:italic>C. sakazakii<\/jats:italic> translocates HBMEC. In this study, the translocation processes of <jats:italic>C. sakazakii<\/jats:italic> on HBMEC were explored. <jats:italic>C. sakazakii<\/jats:italic> strains could effectively adhere to, invade and intracellularly survive in HBMEC. The strain ATCC 29544 exhibited the highest translocation efficiency across HBMEC monolayer among four tested strains. Bacteria-contained intracellular endosomes were detected in <jats:italic>C. sakazakii<\/jats:italic>-infected HBMEC by a transmission electron microscope. Endocytosis-related proteins CD44, Rab5, Rab7, and LAMP2 were increased after infection, while the level of Cathepsin L did not change. <jats:italic>C. sakazakii<\/jats:italic> induced TLR4\/NF-\u03baB inflammatory signal pathway activation in HBMEC, with increased NO production and elevated mRNA levels of IL-8, IL-6, TNF-\u03b1, IL-1\u03b2, iNOS, and COX-2. <jats:italic>C. sakazakii<\/jats:italic> infection also caused LDH release, caspase-3 activation, and HBMEC apoptosis. Meanwhile, increased Dextran-FITC permeability and decreased trans epithelial electric resistance indicated that <jats:italic>C. sakazakii<\/jats:italic> disrupted tight junction of HBMEC monolayers, which was confirmed by the decreased levels of tight junction-related proteins ZO-1 and Occludin. These findings suggest that <jats:italic>C. sakazakii<\/jats:italic> induced intracellular bacterial endocytosis, stimulated inflammation and apoptosis, disrupted monolayer tight junction in HBMEC, which all together contribute to bacterial translocation.<\/jats:p>","DOI":"10.3389\/fmicb.2021.675020","type":"journal-article","created":{"date-parts":[[2021,6,7]],"date-time":"2021-06-07T04:36:27Z","timestamp":1623040587000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":21,"title":["Cronobacter sakazakii ATCC 29544 Translocated Human Brain Microvascular Endothelial Cells via Endocytosis, Apoptosis Induction, and Disruption of Tight Junction"],"prefix":"10.3389","volume":"12","author":[{"given":"Tong","family":"Jin","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ning","family":"Guan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuhang","family":"Du","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinpeng","family":"Zhang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiahui","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaodong","family":"Xia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2021,6,7]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1016\/j.nbd.2009.07.030","article-title":"Structure and function of the blood-brain barrier.","volume":"37","author":"Abbott","year":"2010","journal-title":"Neurobiol. 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