{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,12]],"date-time":"2026-09-12T06:27:24Z","timestamp":1789194444110,"version":"build-2803163510"},"reference-count":47,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2021,4,24]],"date-time":"2021-04-24T00:00:00Z","timestamp":1619222400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000050","name":"National Heart, Lung, and Blood Institute","doi-asserted-by":"publisher","award":["R01HL141105"],"award-info":[{"award-number":["R01HL141105"]}],"id":[{"id":"10.13039\/100000050","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000050","name":"National Heart, Lung, and Blood Institute","doi-asserted-by":"publisher","award":["F31HL145984"],"award-info":[{"award-number":["F31HL145984"]}],"id":[{"id":"10.13039\/100000050","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000968","name":"American Heart Association","doi-asserted-by":"publisher","award":["19PRE34430140"],"award-info":[{"award-number":["19PRE34430140"]}],"id":[{"id":"10.13039\/100000968","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The mechanisms of lymphedema development are not well understood, but emerging evidence highlights the crucial role the immune system plays in driving its progression. It is well known that lymphatic function deteriorates as lymphedema progresses; however, the connection between this progressive loss of function and the immune-driven changes that characterize the disease has not been well established. In this study, we assess changes in leukocyte populations in lymph nodes within the lymphatic drainage basin of the tissue injury site (draining lymph nodes, dLNs) using a mouse tail model of lymphedema in which a pair of draining collecting vessels are left intact. We additionally quantify lymphatic pump function using established near infrared (NIR) lymphatic imaging methods and lymph-draining nanoparticles (NPs) synthesized and employed by our team for lymphatic tissue drug delivery applications to measure lymphatic transport to and resulting NP accumulation within dLNs associated with swelling following surgery. When applied to assess the effects of the anti-inflammatory drug bestatin, which has been previously shown to be a possible treatment for lymphedema, we find lymph-draining NP accumulation within dLNs and lymphatic function to increase as lymphedema progresses, but no significant effect on leukocyte populations in dLNs or tail swelling. These results suggest that ameliorating this loss of lymphatic function is not sufficient to reverse swelling in this surgically induced disease model that better recapitulates the extent of lymphatic injury seen in human lymphedema. It also suggests that loss of lymphatic function during lymphedema may be driven by immune-mediated mechanisms coordinated in dLNs. Our work indicates that addressing both lymphatic vessel dysfunction and immune cell expansion within dLNs may be required to prevent or reverse lymphedema when partial lymphatic function is sustained.<\/jats:p>","DOI":"10.3390\/ijms22094455","type":"journal-article","created":{"date-parts":[[2021,4,24]],"date-time":"2021-04-24T21:49:20Z","timestamp":1619300960000},"page":"4455","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":19,"title":["The Kinetics of Lymphatic Dysfunction and Leukocyte Expansion in the Draining Lymph Node during LTB4 Antagonism in a Mouse Model of Lymphedema"],"prefix":"10.3390","volume":"22","author":[{"given":"Matthew T.","family":"Cribb","sequence":"first","affiliation":[{"name":"Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA 30332, USA"},{"name":"George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0237-2815","authenticated-orcid":false,"given":"Lauren F.","family":"Sestito","sequence":"additional","affiliation":[{"name":"Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1253-474X","authenticated-orcid":false,"given":"Stanley G.","family":"Rockson","sequence":"additional","affiliation":[{"name":"Stanford University School of Medicine, Stanford University, Stanford, CA 94305, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9473-7422","authenticated-orcid":false,"given":"Mark R.","family":"Nicolls","sequence":"additional","affiliation":[{"name":"Stanford University School of Medicine, Stanford University, Stanford, CA 94305, USA"},{"name":"VA Palo Alto Health Care System, Palo Alto, CA 94304, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susan N.","family":"Thomas","sequence":"additional","affiliation":[{"name":"Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA 30332, USA"},{"name":"George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA"},{"name":"Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0351-4587","authenticated-orcid":false,"given":"J. Brandon","family":"Dixon","sequence":"additional","affiliation":[{"name":"Parker H. Petit Institute for Bioengineering and Bioscience, Georgia Institute of Technology, Atlanta, GA 30332, USA"},{"name":"George W. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA"},{"name":"Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA 30332, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,4,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1038\/s41572-019-0072-5","article-title":"Cancer-Associated Secondary Lymphoedema","volume":"5","author":"Rockson","year":"2019","journal-title":"Nat. Rev. Dis. Primers"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Ly, C.L., Kataru, R.P., and Mehrara, B.J. (2017). Inflammatory Manifestations of Lymphedema. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18010171"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"H1065","DOI":"10.1152\/ajpheart.00598.2014","article-title":"Regulation of Inflammation and Fibrosis by Macrophages in Lymphedema","volume":"308","author":"Ghanta","year":"2015","journal-title":"Am. J. Physiol. Heart Circ. Physiol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"706","DOI":"10.1016\/j.jid.2015.12.001","article-title":"Excess Lymphangiogenesis Cooperatively Induced by Macrophages and CD4+ T Cells Drives the Pathogenesis of Lymphedema","volume":"136","author":"Ogata","year":"2016","journal-title":"J. Investig. Dermatol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2193","DOI":"10.1016\/j.ajpath.2016.04.006","article-title":"Prominent Lymphatic Vessel Hyperplasia with Progressive Dysfunction and Distinct Immune Cell Infiltration in Lymphedema","volume":"186","author":"Gousopoulos","year":"2016","journal-title":"Am. J. Pathol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1114","DOI":"10.1096\/fj.12-222695","article-title":"Th2 Differentiation Is Necessary for Soft Tissue Fibrosis and Lymphatic Dysfunction Resulting from Lymphedema","volume":"27","author":"Avraham","year":"2013","journal-title":"FASEB J."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Tabibiazar, R., Cheung, L., Han, J., Swanson, J., Beilhack, A., An, A., Dadras, S.S., Rockson, N., Joshi, S., and Wagner, R. (2006). Inflammatory Manifestations of Experimental Lymphatic Insufficiency. PLoS Med., 3.","DOI":"10.1371\/journal.pmed.0030254"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1172\/jci.insight.89081","article-title":"Regulatory T Cell Transfer Ameliorates Lymphedema and Promotes Lymphatic Vessel Function","volume":"1","author":"Gousopoulos","year":"2016","journal-title":"JCI Insight"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"14345","DOI":"10.1038\/ncomms14345","article-title":"Topical Tacrolimus for the Treatment of Secondary Lymphedema","volume":"8","author":"Gardenier","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Nakamura, K., Radhakrishnan, K., Wong, Y.M., and Rockson, S.G. (2009). Anti-Inflammatory Pharmacotherapy with Ketoprofen Ameliorates Experimental Lymphatic Vascular Insufficiency in Mice. PLoS ONE, 4.","DOI":"10.1371\/journal.pone.0008380"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1006\/abbi.2000.2168","article-title":"Leukotriene B4: Metabolism and Signal Transduction","volume":"385","author":"Yokomizo","year":"2001","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"eaal3920","DOI":"10.1126\/scitranslmed.aal3920","article-title":"Leukotriene B4 Antagonism Ameliorates Experimental Lymphedema","volume":"9","author":"Tian","year":"2017","journal-title":"Sci. Transl. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1970","DOI":"10.1038\/s41467-018-04418-y","article-title":"CD4+ T Cells Are Activated in Regional Lymph Nodes and Migrate to Skin to Initiate Lymphedema","volume":"9","author":"Ly","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"4608","DOI":"10.4049\/jimmunol.1202600","article-title":"Normal Dendritic Cell Mobilization to Lymph Nodes under Conditions of Severe Lymphatic Hypoplasia","volume":"190","author":"Platt","year":"2013","journal-title":"J. Immunol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2181","DOI":"10.4049\/jimmunol.1103545","article-title":"Impaired Humoral Immunity and Tolerance in K14-VEGFR-3-Ig Mice That Lack Dermal Lymphatic Drainage","volume":"189","author":"Thomas","year":"2012","journal-title":"J. Immunol."},{"key":"ref_16","first-page":"118","article-title":"Post-breast cancer lymphedema: Incidence increases from 12 to 30 to 60 months","volume":"43","author":"Armer","year":"2010","journal-title":"Lymphology"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1172\/JCI15830","article-title":"VEGF-C gene therapy augments postnatal lymphangiogenesis and ameliorates secondary lymphedema","volume":"111","author":"Yoon","year":"2003","journal-title":"J. Clin. Investig."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1016\/j.mvr.2006.05.009","article-title":"Secondary Lymphedema in the Mouse Tail: Lymphatic Hyperplasia, VEGF-C Upregulation, and the Protective Role of MMP-9","volume":"72","author":"Rutkowski","year":"2006","journal-title":"Microvasc. Res."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"10405","DOI":"10.1038\/s41598-019-46797-2","article-title":"A Novel Mouse Tail Lymphedema Model for Observing Lymphatic Pump Failure during Lymphedema Development","volume":"9","author":"Weiler","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1463","DOI":"10.1002\/jbm.a.36348","article-title":"S-nitrosated poly(propylene sulfide) nanoparticles for enhanced nitric oxide delivery to lymphatic tissues","volume":"106","author":"Schudel","year":"2018","journal-title":"J. Biomed. Mater. Res. A"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"120411","DOI":"10.1016\/j.biomaterials.2020.120411","article-title":"Lymph-Directed Nitric Oxide Increases Immune Cell Access to Lymph-Borne Nanoscale Solutes","volume":"265","author":"Sestito","year":"2021","journal-title":"Biomaterials"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.jconrel.2006.01.006","article-title":"In Vivo Targeting of Dendritic Cells in Lymph Nodes with Poly(Propylene Sulfide) Nanoparticles","volume":"112","author":"Reddy","year":"2006","journal-title":"J. Control. Release"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"814","DOI":"10.1016\/j.biomaterials.2013.10.003","article-title":"Targeting the Tumor-Draining Lymph Node with Adjuvanted Nanoparticles Reshapes the Anti-Tumor Immune Response","volume":"35","author":"Thomas","year":"2014","journal-title":"Biomaterials"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1038\/s41565-020-0679-4","article-title":"Programmable Multistage Drug Delivery to Lymph Nodes","volume":"15","author":"Schudel","year":"2020","journal-title":"Nat. Nanotechnol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1038\/ni972","article-title":"Leukotriene B4 and BLT1 Control Cytotoxic Effector T Cell Recruitment to Inflamed Tissues","volume":"4","author":"Goodarzi","year":"2003","journal-title":"Nat. Immunol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"371","DOI":"10.1038\/nature12175","article-title":"Neutrophil Swarms Require LTB4 and Integrins at Sites of Cell Death in Vivo","volume":"498","author":"Afonso","year":"2013","journal-title":"Nature"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"066019","DOI":"10.1117\/1.JBO.17.6.066019","article-title":"Sensitivity Analysis of Near-Infrared Functional Lymphatic Imaging","volume":"17","author":"Weiler","year":"2012","journal-title":"J. Biomed. Opt."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"215","DOI":"10.3389\/fphys.2013.00215","article-title":"Differential Transport Function of Lymphatic Vessels in the Rat Tail Model and the Long-Term Effects of Indocyanine Green as Assessed with near-Infrared Imaging","volume":"4","author":"Weiler","year":"2013","journal-title":"Front. Physiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"R281","DOI":"10.1152\/ajpregu.00369.2013","article-title":"Minimally Invasive Method for Determining the Effective Lymphatic Pumping Pressure in Rats Using Near-Infrared Imaging","volume":"306","author":"Nelson","year":"2014","journal-title":"Am. J. Physiol. Regul. Integr. Comp. Physiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"613","DOI":"10.3389\/fimmu.2016.00613","article-title":"T Cell Trafficking through Lymphatic Vessels","volume":"7","author":"Hunter","year":"2016","journal-title":"Front. Immunol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"857","DOI":"10.1016\/j.celrep.2016.12.078","article-title":"T Cell Migration from Inflamed Skin to Draining Lymph Nodes Requires Intralymphatic Crawling Supported by ICAM-1\/LFA-1 Interactions","volume":"18","author":"Teijeira","year":"2017","journal-title":"Cell Rep."},{"key":"ref_32","first-page":"109","article-title":"Trapping of Naive Lymphocytes Triggers Rapid Growth and Remodeling of the Fibroblast Network in Reactive Murine Lymph Nodes","volume":"111","author":"Yang","year":"2014","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"4065","DOI":"10.4049\/jimmunol.1101854","article-title":"Expansion of Cortical and Medullary Sinuses Restrains Lymph Node Hypertrophy during Prolonged Inflammation","volume":"188","author":"Tan","year":"2012","journal-title":"J. Immunol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1146\/annurev-immunol-032713-120252","article-title":"Stromal Cells in Chronic Inflammation and Tertiary Lymphoid Organ Formation","volume":"33","author":"Buckley","year":"2015","journal-title":"Annu. Rev. Immunol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Zampell, J.C., Yan, A., Elhadad, S., Avraham, T., Weitman, E., and Mehrara, B.J. (2012). CD4+ Cells Regulate Fibrosis and Lymphangiogenesis in Response to Lymphatic Fluid Stasis. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0049940"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1016\/j.immuni.2006.01.003","article-title":"B Cell-Driven Lymphangiogenesis in Inflamed Lymph Nodes Enhances Dendritic Cell Mobilization","volume":"24","author":"Angeli","year":"2006","journal-title":"Immunity"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"19817","DOI":"10.1038\/srep19817","article-title":"Inhibition of Inflammation and INOS Improves Lymphatic Function in Obesity","volume":"6","author":"Torrisi","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1007\/s00424-017-1951-9","article-title":"Myeloid Cells Pave the Way for Lymphatic System Development and Maintenance","volume":"469","author":"Ivanov","year":"2017","journal-title":"Pfl\u00fcg. Arch. Eur. J. Physiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1016\/j.immuni.2013.08.007","article-title":"Minimal Differentiation of Classical Monocytes as They Survey Steady-State Tissues and Transport Antigen to Lymph Nodes","volume":"39","author":"Jakubzick","year":"2013","journal-title":"Immunity"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1111\/imm.12478","article-title":"Leukotriene B4-Leukotriene B4 Receptor Axis Promotes Oxazolone-Induced Contact Dermatitis by Directing Skin Homing of Neutrophils and CD8+ T Cells","volume":"146","author":"Lv","year":"2015","journal-title":"Immunology"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1172\/jci.insight.123775","article-title":"Pilot Studies Demonstrate the Potential Benefits of Antiinflammatory Therapy in Human Lymphedema","volume":"3","author":"Rockson","year":"2018","journal-title":"JCI Insight"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"672","DOI":"10.1165\/rcmb.2008-0095OC","article-title":"Leukotriene B4 Release from Mast Cells in IgE-Mediated Airway Hyperresponsiveness and Inflammation","volume":"40","author":"Miyahara","year":"2009","journal-title":"Am. J. Respir. Cell Mol. Biol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1007\/s10059-009-0053-8","article-title":"Leukotriene B4 Regulates Proliferation and Differentiation of Cultured Rat Myoblasts via the BLT1 Pathway","volume":"27","author":"Sun","year":"2009","journal-title":"Mol. Cells"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1903","DOI":"10.1159\/000438551","article-title":"Leukotriene B4 Inhibits L-Type Calcium Channels via P38 Signaling Pathway in Vascular Smooth Muscle Cells","volume":"37","author":"Liu","year":"2015","journal-title":"Cell. Physiol. Biochem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/0090-6980(83)90138-7","article-title":"Effects of arachidonic acid and its cyclo-oxygenase and lipoxygenase products on lymphatic vessel contractility in vitro","volume":"25","author":"Johnston","year":"1983","journal-title":"Prostaglandins"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1016\/0022-4804(88)90046-7","article-title":"Lymphatic smooth muscle responses to leukotrienes, histamine and platelet activating factor","volume":"44","author":"Ferguson","year":"1988","journal-title":"J. Surg. Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"411","DOI":"10.1021\/la0478043","article-title":"Oxidation-Sensitive Polymeric Nanoparticles","volume":"21","author":"Rehor","year":"2005","journal-title":"Langmuir"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/9\/4455\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:52:21Z","timestamp":1760161941000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/9\/4455"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,4,24]]},"references-count":47,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2021,5]]}},"alternative-id":["ijms22094455"],"URL":"https:\/\/doi.org\/10.3390\/ijms22094455","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,4,24]]}}}