{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T20:49:48Z","timestamp":1792788588626,"version":"4.5.0"},"reference-count":83,"publisher":"Public Library of Science (PLoS)","issue":"12","license":[{"start":{"date-parts":[[2021,12,2]],"date-time":"2021-12-02T00:00:00Z","timestamp":1638403200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"ILIAS Biologics & ILIAS Therapeutics, Inc., USA"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"\n Background<\/jats:title>\n Microbial invasion of the intraamniotic cavity and intraamniotic inflammation are factors associated with spontaneous preterm birth. Understanding the route and kinetics of infection, sites of colonization, and mechanisms of host inflammatory response is critical to reducing preterm birth risk.<\/jats:p>\n <\/jats:sec>\n \n Objectives<\/jats:title>\n \n This study developed an animal model of ascending infection and preterm birth with live bacteria (\n E<\/jats:italic>\n .\n coli<\/jats:italic>\n ) in pregnant CD-1 mice with the goal of better understanding the process of microbial invasion of the intraamniotic cavity and intraamniotic inflammation.\n <\/jats:p>\n <\/jats:sec>\n \n Study design<\/jats:title>\n \n Multiple experiments were conducted in this study. To determine the dose of\n E<\/jats:italic>\n .\n coli<\/jats:italic>\n required to induce preterm birth, CD-1 mice were injected vaginally with four different doses of\n E<\/jats:italic>\n .\n coli<\/jats:italic>\n (10\n 3<\/jats:sup>\n , 10\n 6<\/jats:sup>\n , 10\n 10<\/jats:sup>\n , or 10\n 11<\/jats:sup>\n colony forming units [CFU]) in 40 \u03bcL of nutrient broth or broth alone (control) on an embryonic day (E)15. Preterm birth (defined as delivery before E18.5) was monitored using live video.\n E<\/jats:italic>\n .\n coli<\/jats:italic>\n ascent kinetics were measured by staining the\n E<\/jats:italic>\n .\n coli<\/jats:italic>\n with lipophilic tracer DiD for visualization through intact tissue with an in vivo imaging system (IVIS) after inoculation. The\n E<\/jats:italic>\n .\n coli<\/jats:italic>\n were also directly visualized in reproductive tissues by staining the bacteria with carboxyfluorescein succinimidyl ester (CFSE) prior to administration and via immunohistochemistry (IHC) by staining tissues with anti-\n E<\/jats:italic>\n .\n coli<\/jats:italic>\n antibody. Each pup\u2019s amniotic fluid was cultured separately to determine the extent of microbial invasion of the intraamniotic cavity at different time points. Intraamniotic inflammation resulting from\n E<\/jats:italic>\n .\n coli<\/jats:italic>\n invasion was assessed with IHC for inflammatory markers (TLR-4, P-NF-\u03baB) and neutrophil marker (Ly-6G) for chorioamnionitis at 6- and 24-h post-inoculation.\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n Vaginally administered\n E<\/jats:italic>\n .\n coli<\/jats:italic>\n resulted in preterm birth in a dose-dependent manner with higher doses causing earlier births. In\n ex vivo<\/jats:italic>\n imaging and IHC detected uterine horns proximal to the cervix had increased\n E<\/jats:italic>\n .\n coli<\/jats:italic>\n compared to the distal uterine horns.\n E<\/jats:italic>\n .\n coli<\/jats:italic>\n were detected in the uterus, fetal membranes (FM), and placenta in a time-dependent manner with 6 hr having increased intensity of\n E<\/jats:italic>\n .\n coli<\/jats:italic>\n positive signals in pups near the cervix and in all pups at 24 hr. Similarly,\n E<\/jats:italic>\n .\n coli<\/jats:italic>\n grew from the cultures of amniotic fluid collected nearest to the cervix, but not from the more distal samples at 6 hr post-inoculation. At 24 hr, all amniotic fluid cultures regardless of distance from the cervix, were positive for\n E<\/jats:italic>\n .\n coli<\/jats:italic>\n . TLR-4 and P-NF-\u03baB signals were more intense in the tissues where\n E<\/jats:italic>\n .\n coli<\/jats:italic>\n was present (placenta, FM and uterus), displaying a similar trend toward increased signal in proximal gestational sacs compared to distal at 6 hr. Ly-6G+ cells, used to confirm chorioamnionitis, were increased at 24 hr compared to 6 hr post-inoculation and control.\n <\/jats:p>\n <\/jats:sec>\n \n Conclusion<\/jats:title>\n We report the development of mouse model of ascending infection and the associated inflammation of preterm birth. Clinically, these models can help to understand mechanisms of infection associated preterm birth, determine targets for intervention, or identify potential biomarkers that can predict a high-risk pregnancy status early in pregnancy.<\/jats:p>\n <\/jats:sec>","DOI":"10.1371\/journal.pone.0260370","type":"journal-article","created":{"date-parts":[[2021,12,2]],"date-time":"2021-12-02T14:05:19Z","timestamp":1638453919000},"page":"e0260370","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":34,"title":["Development of a mouse model of ascending infection and preterm birth"],"prefix":"10.1371","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5141-6223","authenticated-orcid":true,"given":"Nicholas R.","family":"Spencer","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8142-8137","authenticated-orcid":true,"given":"Enkhtuya","family":"Radnaa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tuvshintugs","family":"Baljinnyam","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Talar","family":"Kechichian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4535-8837","authenticated-orcid":true,"given":"Ourlad Alzeus G.","family":"Tantengco","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elizabeth","family":"Bonney","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ananth Kumar","family":"Kammala","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3991-1747","authenticated-orcid":true,"given":"Samantha","family":"Sheller-Miller","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9213-6105","authenticated-orcid":true,"given":"Ramkumar","family":"Menon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2021,12,2]]},"reference":[{"issue":"1","key":"pone.0260370.ref001","doi-asserted-by":"crossref","first-page":"31","DOI":"10.2471\/BLT.08.062554","article-title":"The worldwide incidence of preterm birth: a systematic review of maternal mortality and morbidity","volume":"88","author":"S Beck","year":"2010","journal-title":"Bull World Health Organ"},{"issue":"6","key":"pone.0260370.ref002","doi-asserted-by":"crossref","first-page":"590","DOI":"10.1080\/00016340802005126","article-title":"Spontaneous preterm birth, a clinical dilemma: etiologic, pathophysiologic and genetic heterogeneities and racial disparity","volume":"87","author":"R. 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