New Model of Macrophage Acquisition of the Lymphatic Endothelial Phenotype 英文参考文献.docVIP

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New Model of Macrophage Acquisition of the Lymphatic Endothelial Phenotype 英文参考文献.doc

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New Model of Macrophage Acquisition of the Lymphatic Endothelial Phenotype 英文参考文献

NewModelofMacrophageAcquisitionoftheLymphatic EndothelialPhenotype KellyL.Hall.,LisaD.Volk-Draper.,MichaelJ.Flister.,SophiaRan* DepartmentofMedicalMicrobiology,Immunology,andCellBiology,SouthernIllinoisUniversitySchoolofMedicine,Springfield,Illinois,UnitedStatesofAmerica Abstract Background: Macrophage-derived lymphatic endothelial cell progenitors (M-LECPs) contribute to new lymphatic vessel formation,butthemechanismsregulatingtheirdifferentiation,recruitment,andfunctionarepoorlyunderstood.Detailed characterization of M-LECPs is limited by low frequency in vivo and lack of model systems allowing in-depth molecular analysesinvitro.Ourgoalwastoestablishacellculturemodeltocharacterizeinflammation-inducedmacrophage-to-LECP differentiationundercontrolledconditions. Methodology/PrincipalFindings:Time-courseanalysisofdiaphragmsfromlipopolysaccharide(LPS)-treatedmicerevealed rapidmobilizationofbonemarrow-derivedandperitonealmacrophagestotheproximityoflymphaticvesselsfollowedby widespread (,50%)incorporation of M-LECPs intotheinflamedlymphatic vasculature.Adifferentiation shift toward the lymphaticphenotypewasfoundinthreeLPS-inducedsubsetsofactivatedmacrophagesthatwerepositiveforVEGFR-3and manyotherlymphatic-specificmarkers.VEGFR-3wasstronglyelevatedintheearlystageofmacrophagetransitiontoLECPs but undetectable in M-LECPs prior to vascular integration. Similar transient pattern of VEGFR-3 expression was found in RAW264.7macrophagesactivatedbyLPSinvitro.ActivatedRAW264.7cellsco-expressedVEGF-Cthatinducedanautocrine signaling loop as indicated by VEGFR-3 phosphorylation inhibited by a soluble receptor. LPS-activated RAW264.7 macrophagesalsoshoweda68%overlapwithendogenousCD11b+/VEGFR-3+LECPsintheexpressionoflymphatic-specific genes.Moreover,wheninjectedintoLPS-butnotsaline-treatedmice,GFP-taggedRAW264.7cellsmassivelyinfiltratedthe inflameddiaphragmfollowedbyintegrationinto18%oflymphaticvessels. Conclusions/Significance: We present a new model for macrophage-LECP differentiat