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Cell. Infect. Microbiol."],"abstract":"<jats:p><jats:italic>Salmonella<\/jats:italic> Enteritidis is an important intracellular pathogen, which can cause gastroenteritis in humans and animals and threaten life and health. <jats:italic>S.<\/jats:italic> Enteritidis proliferates in host macrophages to establish systemic infection. In this study, we evaluated the effects of <jats:italic>Salmonella<\/jats:italic> pathogenicity island-1 (SPI-1) and SPI-2 to <jats:italic>S.<\/jats:italic> Enteritidis virulence <jats:italic>in vitro<\/jats:italic> and <jats:italic>in vivo<\/jats:italic>, as well as the host inflammatory pathways affected by SPI-1 and SPI-2. Our results show that <jats:italic>S.<\/jats:italic> Enteritidis SPI-1 and SPI-2 contributed to bacterial invasion and proliferation in RAW264.7 macrophages, and induced cytotoxicity and cellular apoptosis of these cells. <jats:italic>S.<\/jats:italic> Enteritidis infection induced multiple inflammatory responses, including mitogen-activated protein kinase (ERK-mediated) and Janus kinase-signal transducer and activator of transcript (STAT) (STAT2-mediated) pathways. Both SPI-1 and SPI-2 were necessary to induce robust inflammatory responses and ERK\/STAT2 phosphorylation in macrophages. In a mouse infection model, both SPIs, especially SPI-2, resulted in significant production of inflammatory cytokines and various interferon-stimulated genes in the liver and spleen. Activation of the ERK- and STAT2-mediated cytokine storm was largely affected by SPI-2. <jats:italic>S.<\/jats:italic> Enteritidis <jats:italic>\u0394SPI-1<\/jats:italic>-infected mice displayed moderate histopathological damage and drastically reduced bacterial loads in tissues, whereas only slight damage and no bacteria were observed in <jats:italic>\u0394SPI-2-<\/jats:italic> and <jats:italic>\u0394SPI-1\/SPI-2<\/jats:italic>-infected mice. A survival assay showed that <jats:italic>\u0394SPI-1<\/jats:italic> mutant mice maintained a medium level of virulence, while SPI-2 plays a decisive role in bacterial virulence. Collectively, our findings indicate that both SPIs, especially SPI-2, profoundly contributed to <jats:italic>S.<\/jats:italic> Enteritidis intracellular localization and virulence by activating multiple inflammatory pathways.<\/jats:p>","DOI":"10.3389\/fcimb.2023.1158888","type":"journal-article","created":{"date-parts":[[2023,5,31]],"date-time":"2023-05-31T14:04:48Z","timestamp":1685541888000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":27,"title":["Salmonella Enteritidis activates inflammatory storm via SPI-1 and SPI-2 to promote intracellular proliferation and bacterial virulence"],"prefix":"10.3389","volume":"13","author":[{"given":"Dan","family":"Xiong","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Li","family":"Song","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yushan","family":"Chen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xinan","family":"Jiao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiming","family":"Pan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,5,30]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"197","DOI":"10.1016\/j.cell.2013.06.013","article-title":"A systems analysis identifies a feedforward inflammatory circuit leading to lethal influenza infection","volume":"154","author":"Brandes","year":"2013","journal-title":"Cell"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.0010032","article-title":"Salmonella pathogenicity island 2 is expressed prior to penetrating the intestine","volume":"1","author":"Brown","year":"2005","journal-title":"PloS Pathog."},{"key":"B3","unstructured":"Salmonella2023"},{"key":"B4","doi-asserted-by":"publisher","first-page":"2043","DOI":"10.1038\/sj.emboj.7600676","article-title":"Formation of a novel surface structure encoded by Salmonella pathogenicity island 2","volume":"24","author":"Chakravortty","year":"2005","journal-title":"EMBO J."},{"key":"B5","doi-asserted-by":"publisher","first-page":"6640","DOI":"10.1073\/pnas.120163297","article-title":"One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products","volume":"97","author":"Datsenko","year":"2000","journal-title":"Proc. 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