{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T05:10:55Z","timestamp":1790745055659,"version":"4.1.0"},"reference-count":39,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2019,2,27]],"date-time":"2019-02-27T00:00:00Z","timestamp":1551225600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2019,2,27]],"date-time":"2019-02-27T00:00:00Z","timestamp":1551225600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100001691","name":"MEXT | Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["JP17K16985"],"award-info":[{"award-number":["JP17K16985"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Lymphatic vessels play a crucial role in systemic immune response and regulation of tissue fluid homeostasis. Corneal lymphangiogenesis in bacterial keratitis has not been studied. In this study, we investigated the mechanism and the role of corneal lymphangiogenesis in a murine bacterial keratitis model using <jats:italic>Pseudomonas aeruginosa<\/jats:italic>. We first demonstrated that corneal lymphangiogenesis was enhanced mainly in the late stage of bacterial keratitis, contrary to corneal angiogenesis that started earlier. Corresponding to the delayed lymphangiogenesis, expression of the pro-lymphangiogenic factors VEGF-C and VEGFR-3 increased in the late stage of bacterial keratitis. We further found that F4\/80 and CD11b positive macrophages played an essential role in corneal lymphangiogenesis. Notably, macrophages were specifically involved in corneal lymphangiogenesis in the late stage of bacterial keratitis. Finally, we demonstrated the beneficial role of corneal lymphangiogenesis in ameliorating the clinical course of bacterial keratitis. Our study showed that bacterial activity was not directly involved in the late stage of keratitis, while corneal lymphangiogenesis reduced corneal edema and clinical manifestation in the late stage of bacterial keratitis. These findings suggest that the process of lymphangiogenesis in bacterial keratitis ameliorates corneal inflammation and edema in the late stage of bacterial keratitis.<\/jats:p>","DOI":"10.1038\/s41598-019-39876-x","type":"journal-article","created":{"date-parts":[[2019,2,27]],"date-time":"2019-02-27T11:27:43Z","timestamp":1551266863000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":34,"title":["Corneal lymphangiogenesis ameliorates corneal inflammation and edema in late stage of bacterial keratitis"],"prefix":"10.1038","volume":"9","author":[{"given":"Akitomo","family":"Narimatsu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takaaki","family":"Hattori","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naohito","family":"Koike","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2663-471X","authenticated-orcid":false,"given":"Kazuki","family":"Tajima","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hayate","family":"Nakagawa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naoyuki","family":"Yamakawa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoshihiko","family":"Usui","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shigeto","family":"Kumakura","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tetsuya","family":"Matsumoto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiroshi","family":"Goto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2019,2,27]]},"reference":[{"key":"39876_CR1","doi-asserted-by":"publisher","first-page":"1371","DOI":"10.1038\/nm.2545","volume":"17","author":"K Alitalo","year":"2011","unstructured":"Alitalo, K. The lymphatic vasculature in disease. Nature Medicine 17, 1371\u20131380 (2011).","journal-title":"Nature Medicine"},{"key":"39876_CR2","doi-asserted-by":"publisher","first-page":"11658","DOI":"10.1073\/pnas.0404272101","volume":"101","author":"LK Chang","year":"2004","unstructured":"Chang, L. K. et al. Dose-dependent response of FGF-2 for lymphangiogenesis. Proc. Natl. Acad. Sci. 101, 11658\u201311663 (2004).","journal-title":"Proc. Natl. Acad. Sci."},{"key":"39876_CR3","doi-asserted-by":"publisher","first-page":"228","DOI":"10.4161\/cc.4.2.1421","volume":"4","author":"Y Cao","year":"2005","unstructured":"Cao, Y. Direct role of PDGF-BB in lymphangiogenesis and lymphatic metastasis. Cell Cycle 4, 228\u2013230 (2005).","journal-title":"Cell Cycle"},{"key":"39876_CR4","doi-asserted-by":"publisher","first-page":"1040","DOI":"10.1172\/JCI20465","volume":"113","author":"C Cursiefen","year":"2004","unstructured":"Cursiefen, C. et al. VEGF-A stimulates lymphangiogenesis and hemangiogenesis in inflammatory neovascularization via macrophage recruitment. J. Clin. Invest. 113, 1040\u20131050 (2004).","journal-title":"J. Clin. Invest."},{"key":"39876_CR5","doi-asserted-by":"publisher","first-page":"826","DOI":"10.1016\/j.bbrc.2008.10.077","volume":"377","author":"K Watari","year":"2008","unstructured":"Watari, K. et al. Role of macrophages in inflammatory lymphangiogenesis: Enhanced production of vascular endothelial growth factor C and D through NF-\u03baB activation. Biochem. Biophys. Res. Commun. 377, 826\u2013831 (2008).","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"39876_CR6","doi-asserted-by":"publisher","first-page":"1423","DOI":"10.1126\/science.276.5317.1423","volume":"276","author":"M Jeltsch","year":"1997","unstructured":"Jeltsch, M. et al. Hyperplasia of lymphatic vessels in VFGF transgenic mouse. Science 276, 1423\u20131425 (1997).","journal-title":"Science"},{"key":"39876_CR7","doi-asserted-by":"publisher","first-page":"74","DOI":"10.1038\/ni1013","volume":"5","author":"MJ Karkkainen","year":"2004","unstructured":"Karkkainen, M. J. et al. Vascular endothelial growth factor C is required for sprouting of the first lymphatic vessels from embryonic veins. Nat. Immunol. 5, 74\u201380 (2004).","journal-title":"Nat. Immunol."},{"key":"39876_CR8","doi-asserted-by":"publisher","first-page":"210","DOI":"10.1038\/nrclinonc.2011.21","volume":"8","author":"JML Ebos","year":"2011","unstructured":"Ebos, J. M. L. & Kerbel, R. S. Antiangiogenic therapy: impact on invasion, disease progression, and metastasis. Nat. Rev. Clin. Oncol. 8, 210\u2013221 (2011).","journal-title":"Nat. Rev. Clin. Oncol."},{"key":"39876_CR9","doi-asserted-by":"publisher","first-page":"669","DOI":"10.1038\/nm0603-669","volume":"9","author":"N Ferrara","year":"2003","unstructured":"Ferrara, N., Gerber, H. P. & LeCouter, J. The biology of VEGF and its receptors. Nature Medicine 9, 669\u2013676 (2003).","journal-title":"Nature Medicine"},{"key":"39876_CR10","doi-asserted-by":"publisher","first-page":"154","DOI":"10.1016\/j.ceb.2008.12.012","volume":"21","author":"M Lohela","year":"2009","unstructured":"Lohela, M., Bry, M., Tammela, T. & Alitalo, K. VEGFs and receptors involved in angiogenesis versus lymphangiogenesis. Current Opinion in Cell Biology 21, 154\u2013165 (2009).","journal-title":"Current Opinion in Cell Biology"},{"key":"39876_CR11","doi-asserted-by":"publisher","first-page":"993","DOI":"10.1038\/nature05249","volume":"443","author":"BK Ambati","year":"2006","unstructured":"Ambati, B. K. et al. Corneal avascularity is due to soluble VEGF receptor-1. Nature 443, 993\u2013997 (2006).","journal-title":"Nature"},{"key":"39876_CR12","first-page":"2127","volume":"43","author":"C Cursiefen","year":"2002","unstructured":"Cursiefen, C. et al. Lymphatic vessels in vascularized human corneas: immunohistochemical investigation using LYVE-1 and podoplanin. Invest. Ophthalmol. Vis. Sci. 43, 2127\u20132135 (2002).","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"39876_CR13","doi-asserted-by":"publisher","first-page":"2363","DOI":"10.1172\/JCI23874","volume":"115","author":"K Maruyama","year":"2005","unstructured":"Maruyama, K. et al. Inflammation-induced lymphangiogenesis in the cornea arises from CD11b-positive macrophages. J. Clin. Invest. 115, 2363\u20132372 (2005).","journal-title":"J. Clin. Invest."},{"key":"39876_CR14","doi-asserted-by":"publisher","first-page":"1081","DOI":"10.1182\/blood-2010-02-267427","volume":"117","author":"S Nakao","year":"2011","unstructured":"Nakao, S. et al. Blood vessel endothelial VEGFR-2 delays lymphangiogenesis: an endogenous trapping mechanism links lymph- and angiogenesis. Blood 117, 1081\u20131090 (2011).","journal-title":"Blood"},{"key":"39876_CR15","doi-asserted-by":"publisher","first-page":"315","DOI":"10.1111\/j.1755-3768.2008.01349.x","volume":"87","author":"S Ling","year":"2009","unstructured":"Ling, S. et al. Development of new lymphatic vessels in alkali-burned corneas. Acta Ophthalmologica 87, 315\u2013322 (2009).","journal-title":"Acta Ophthalmologica"},{"key":"39876_CR16","doi-asserted-by":"publisher","first-page":"3082","DOI":"10.1167\/iovs.12-10856","volume":"54","author":"B Detry","year":"2013","unstructured":"Detry, B. et al. Sunitinib inhibits inflammatory corneal lymphangiogenesis. Invest. Ophthalmol. Vis. Sci. 54, 3082\u20133093 (2013).","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"39876_CR17","doi-asserted-by":"publisher","first-page":"101","DOI":"10.1084\/jem.20091385","volume":"207","author":"TR Wuest","year":"2010","unstructured":"Wuest, T. R. & Carr, D. J. VEGF-A expression by HSV-1-infected cells drives corneal lymphangiogenesis. J. Exp. Med. 207, 101\u2013115 (2010).","journal-title":"J. Exp. Med."},{"key":"39876_CR18","doi-asserted-by":"publisher","first-page":"60","DOI":"10.1016\/j.survophthal.2014.06.001","volume":"60","author":"PJ Park","year":"2015","unstructured":"Park, P. J. et al. Corneal lymphangiogenesis in herpetic stromal keratitis. Surv. Ophthalmol. 60, 60\u201371 (2015).","journal-title":"Surv. Ophthalmol."},{"key":"39876_CR19","doi-asserted-by":"publisher","first-page":"84","DOI":"10.1001\/archophthalmol.2011.266","volume":"130","author":"S Goyal","year":"2012","unstructured":"Goyal, S., Chauhan, S. K. & Dana, R. Blockade of prolymphangiogenic vascular endothelial growth factor C in dry eye disease. Arch. Ophthalmol. 130, 84\u201389 (2012).","journal-title":"Arch. Ophthalmol."},{"key":"39876_CR20","doi-asserted-by":"publisher","first-page":"535","DOI":"10.4049\/jimmunol.0903180","volume":"184","author":"T Dietrich","year":"2010","unstructured":"Dietrich, T. et al. Cutting edge: lymphatic vessels, not blood vessels, primarily mediate immune rejections after transplantation. J. Immunol. 184, 535\u2013539 (2010).","journal-title":"J. Immunol."},{"key":"39876_CR21","doi-asserted-by":"publisher","first-page":"1755","DOI":"10.1007\/s00417-014-2749-5","volume":"252","author":"P Emami-Naeini","year":"2014","unstructured":"Emami-Naeini, P. et al. Soluble vascular endothelial growth factor receptor-3 suppresses allosensitization and promotes corneal allograft survival. Graefe\u2019s Arch. Clin. Exp. Ophthalmol. 252, 1755\u20131762 (2014).","journal-title":"Graefe\u2019s Arch. Clin. Exp. Ophthalmol."},{"key":"39876_CR22","doi-asserted-by":"publisher","first-page":"3140","DOI":"10.1167\/iovs.14-16186","volume":"56","author":"HS Lee","year":"2015","unstructured":"Lee, H. S. et al. Involvement of corneal lymphangiogenesis in a mouse model of allergic eye disease. Invest. Ophthalmol. Vis. Sci. 56, 3140\u20133148 (2015).","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"39876_CR23","doi-asserted-by":"crossref","unstructured":"Hos, D. et al. Transient ingrowth of lymphatic vessels into the physiologically avascular cornea regulates corneal edema and transparency. Sci. Rep. 7 (2017).","DOI":"10.1038\/s41598-017-07806-4"},{"key":"39876_CR24","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1006\/exer.2000.0925","volume":"72","author":"T Mimura","year":"2001","unstructured":"Mimura, T. et al. Expression of vascular endothelial growth factor C and vascular endothelial growth factor receptor 3 in corneal lymphangiogenesis. Exp. Eye Res. 72, 71\u201378 (2001).","journal-title":"Exp. Eye Res."},{"key":"39876_CR25","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/j.preteyeres.2003.10.002","volume":"23","author":"LD Hazlett","year":"2004","unstructured":"Hazlett, L. D. Corneal response to Pseudomonas aeruginosa infection. Prog. Retin. Eye Res. 23, 1\u201330 (2004).","journal-title":"Prog. Retin. Eye Res."},{"key":"39876_CR26","doi-asserted-by":"publisher","first-page":"185","DOI":"10.1038\/eye.2011.288","volume":"26","author":"F Stapleton","year":"2012","unstructured":"Stapleton, F. & Carnt, N. Contact lens-related microbial keratitis: How have epidemiology and genetics helped us with pathogenesis and prophylaxis. Eye 26, 185\u2013193 (2012).","journal-title":"Eye"},{"key":"39876_CR27","doi-asserted-by":"publisher","first-page":"4910","DOI":"10.1167\/iovs.06-0537","volume":"47","author":"X Huang","year":"2006","unstructured":"Huang, X., Du, W., McClellan, S. A., Barrett, R. P. & Hazlett, L. D. TLR4 is required for host resistance in Pseudomonas aeruginosa keratitis. Invest. Ophthalmol. Vis. Sci. 47, 4910\u20134916 (2006).","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"39876_CR28","doi-asserted-by":"publisher","first-page":"4260","DOI":"10.3390\/md12074260","volume":"12","author":"R Nijland","year":"2014","unstructured":"Nijland, R., Hofland, T. & Van Strijp, J. A. G. Recognition of LPS by TLR4: Potential for anti-inflammatory therapies. Marine Drugs 12, 4260\u20134273 (2014).","journal-title":"Marine Drugs"},{"key":"39876_CR29","doi-asserted-by":"crossref","first-page":"73711","DOI":"10.18632\/oncotarget.12449","volume":"7","author":"G Zhu","year":"2016","unstructured":"Zhu, G. et al. Lipopolysaccharide increases the release of VEGF-C that enhances cell motility and promotes lymphangiogenesis and lymphatic metastasis through the TLR4- NF-\u03baB\/JNK pathways in colorectal cancer. Oncotarget 7, 73711\u201373724 (2016).","journal-title":"Oncotarget"},{"key":"39876_CR30","doi-asserted-by":"publisher","first-page":"2605","DOI":"10.1182\/blood-2008-07-166934","volume":"113","author":"S Kang","year":"2009","unstructured":"Kang, S. et al. Toll-like receptor 4 in lymphatic endothelial cells contributes to LPS-induced lymphangiogenesis by chemotactic recruitment of macrophages. Blood 113, 2605\u20132613 (2009).","journal-title":"Blood"},{"key":"39876_CR31","doi-asserted-by":"publisher","first-page":"5219","DOI":"10.4049\/jimmunol.170.10.5219","volume":"170","author":"SA McClellan","year":"2003","unstructured":"McClellan, S. A., Huang, X., Barrett, R. P., van Rooijen, N. & Hazlett, L. D. Macrophages restrict Pseudomonas aeruginosa growth, regulate polymorphonuclear neutrophil influx, and balance pro- and anti-inflammatory cytokines in BALB\/c mice. J Immunol 170, 5219\u20135227 (2003).","journal-title":"J Immunol"},{"key":"39876_CR32","doi-asserted-by":"publisher","first-page":"1506","DOI":"10.1167\/iovs.16-19134","volume":"57","author":"CK Muraleedharan","year":"2016","unstructured":"Muraleedharan, C. K. et al. Inactivation of the miR-183\/96\/182 cluster decreases the severity of Pseudomonas aeruginosa-induced keratitis. Invest. Ophthalmol. Vis. Sci. 57, 1506\u20131517 (2016).","journal-title":"Invest. Ophthalmol. Vis. Sci."},{"key":"39876_CR33","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1016\/j.ajpath.2015.09.012","volume":"186","author":"D Hos","year":"2016","unstructured":"Hos, D. et al. IL-10 Indirectly regulates corneal lymphangiogenesis and resolution of inflammation via macrophages. Am. J. Pathol. 186, 159\u2013171 (2016).","journal-title":"Am. J. Pathol."},{"key":"39876_CR34","doi-asserted-by":"publisher","first-page":"4272","DOI":"10.4049\/jimmunol.1000874","volume":"185","author":"Y Sun","year":"2010","unstructured":"Sun, Y. et al. TLR4 and TLR5 on Corneal macrophages regulate Pseudomonas aeruginosa keratitis by signaling through MyD88-dependent and -independent pathways. J. Immunol. 185, 4272\u20134283 (2010).","journal-title":"J. Immunol."},{"key":"39876_CR35","doi-asserted-by":"publisher","first-page":"567","DOI":"10.1146\/annurev-physiol-022516-034348","volume":"79","author":"K Hamidzadeh","year":"2017","unstructured":"Hamidzadeh, K., Christensen, S. M., Dalby, E., Chandrasekaran, P. & Mosser, D. M. Macrophages and the recovery from acute and chronic inflammation. Annu. Rev. Physiol. 79, 567\u2013592 (2017).","journal-title":"Annu. Rev. Physiol."},{"key":"39876_CR36","doi-asserted-by":"publisher","first-page":"412","DOI":"10.1016\/j.cmet.2016.11.009","volume":"25","author":"Y Oishi","year":"2017","unstructured":"Oishi, Y. et al. SREBP1 Contributes to resolution of pro-inflammatory TLR4 signaling by reprogramming fatty acid metabolism. Cell Metab. 25, 412\u2013427 (2017).","journal-title":"Cell Metab."},{"key":"39876_CR37","doi-asserted-by":"publisher","first-page":"445","DOI":"10.1001\/archophthalmol.2011.42","volume":"129","author":"D Hos","year":"2011","unstructured":"Hos, D. et al. Suppression of inflammatory corneal lymphangiogenesis by application of topical corticosteroids. Arch. Ophthalmol. 129, 445\u2013452 (2011).","journal-title":"Arch. Ophthalmol."},{"key":"39876_CR38","doi-asserted-by":"publisher","first-page":"953","DOI":"10.1038\/nri1733","volume":"5","author":"S Gordon","year":"2005","unstructured":"Gordon, S. & Taylor, P. R. Monocyte and macrophage heterogeneity. Nat. Rev. Immunol. 5, 953\u2013964 (2005).","journal-title":"Nat. Rev. Immunol."},{"key":"39876_CR39","first-page":"189","volume":"280","author":"J Biewenga","year":"1995","unstructured":"Biewenga, J. et al. Macrophage depletion in the rat after intraperitoneal administration of liposome-encapsulated clodronate: depletion kinetics and accelerated repopulation of peritoneal and omental macrophages by administration of Freund\u2019s adjuvant. Cell Tissue Res. 280, 189\u2013196 (1995).","journal-title":"Cell Tissue Res."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-39876-x.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-39876-x","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-39876-x.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,17]],"date-time":"2022-12-17T06:38:11Z","timestamp":1671259091000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-39876-x"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,2,27]]},"references-count":39,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2019,12]]}},"alternative-id":["39876"],"URL":"https:\/\/doi.org\/10.1038\/s41598-019-39876-x","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,2,27]]},"assertion":[{"value":"11 November 2018","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"1 February 2019","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"27 February 2019","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"2984"}}