Technical Advance: Live‐imaging analysis of human dendritic cell migrating behavior under the influence of immune‐stimulating reagents in an organotypic model of lung

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

关键词: 3D‐;    tissue models;    epithelial tissue;    Toll‐;    like receptor agonists;    inflammation;   

DOI  :  10.1189/jlb.3TA0513-303R

学科分类：生理学

来源: Federation of American Societies for Experimental Biology

【 摘 要 】

ThismanuscriptdescribestechnicaladvancesallowingmanipulationandquantitativeanalysesofhumanDCmigratorybehaviorinlungepithelialtissue.DCsarehematopoieticcellsessentialforthemaintenanceoftissuehomeostasisandtheinductionoftissue‐specificimmuneresponses.Importantfunctionsincludecytokineproductionandmigrationinresponsetoinfectionfortheinductionofproperimmuneresponses.TodesignappropriatestrategiestoexploithumanDCfunctionalpropertiesinlungtissueforthepurposeofclinicalevaluation,e.g.,candidatevaccinationandimmunotherapystrategies,wehavedevelopedalive‐imagingassaybasedonourpreviouslydescribedorganotypicmodelofthehumanlung.ThisassayallowsprovocationsandsubsequentquantitativeinvestigationsofDCfunctionalpropertiesunderconditionsmimickingmorphologicalandfunctionalfeaturesoftheinvivoparentaltissue.Wepresentprotocolstosetupandpreparetissuemodelsfor4D(x,y,z,time)fluorescence‐imaginganalysisthatallowspatialandtemporalstudiesofhumanDCsinliveepithelialtissue,followedbyflowcytometryanalysisofDCsretrievedfromdigestedtissuemodels.ThismodelsystemcanbeusefulforelucidatingincompletelydefinedpathwayscontrollingDCfunctionalresponsestoinfectionandinflammationinlungepithelialtissue,aswellastheefficacyoflocallyadministeredcandidateinterventions...

【 授权许可】

CC BY