A novel microfluidic assay reveals a key role for protein kinase C δ in regulating human neutrophil–endothelium interaction

Journal of Leukocyte Biology: An Official Publication of the Reticuloendothelial Society

关键词: inflammation;    adhesion;    biomimetic;    leukocytes;    transmigration;   

DOI  :  10.1189/jlb.3MA0216-087R

学科分类：生理学

来源: Federation of American Societies for Experimental Biology

【 摘 要 】

Akeystepinneutrophil‐mediatedtissuedamageisthemigrationofactivatedneutrophilsacrossthevascularendothelium.Previously,weidentifiedproteinkinaseCδasacriticalregulatorofneutrophilmigrationinsepsisbutdidnotidentifyspecificstepsinmigration.Inthisstudy,weusedournovelbiomimeticmicrofluidicassaytodelineatesystematicallythemechanismbywhichproteinkinaseCδregulatesindividualstepsinhumanneutrophil–endothelialinteractionduringinflammation.Thebiomimeticmicrofluidicassayincludesanetworkofvascularchannels,producedfrominvivoimagesconnectedtoatissuecompartmentthroughaporousbarrier.HUVECsculturedinvascularchannelsformedacompletelumenunderphysiologicshearflow.HUVECswerepretreatedwithTNF‐α±aproteinkinaseCδinhibitor,andthetissuecompartmentwasfilledwithachemoattractant(fMLPorIL‐8).Underphysiologicshearflow,theroleofproteinkinaseCδonspatialandtemporalneutrophiladherence/migrationwasquantified.ProteinkinaseCδinhibitionsignificantlyreducedneutrophiladhesioninresponsetofMLPandIL‐8onlyunderlowshearrateandnearbifurcations.ProteinkinaseCδinhibitionalsodecreasedadherencetononactivatedHUVECsinresponsetofMLPorIL‐8.ProteinkinaseCδinhibitionreducedneutrophilmigrationintothetissuecompartmentinresponsetofMLPandtoalesserdegree,toIL‐8.Antibody‐coatedmicroparticlesdemonstratedthatproteinkinaseCδinhibitiondown‐regulatedE‐selectinandICAM‐1butnotVCAM‐1expression.Withtheuseofaphysiologicallyrelevantinvitromodelsystem,wedemonstratethatproteinkinaseCδplaysanimportantroleintheregulationofneutrophiladherence/migrationduringinflammationandidentifieskeystepsregulatedbyproteinkinaseCδinneutrophil–endothelialinteractions...

【 授权许可】

CC BY