title: A short protocol using dexamethasone and monophosphoryl lipid A generates tolerogenic dendritic cells that display a potent migratory capacity to lymphoid chemokines.
abstract: BACKGROUND
  Generation of tolerogenic dendritic cells (TolDCs) for therapy is challenging due to its implications for the design of protocols suitable for clinical applications, which means not only using safe products, but also working at defining specific biomarkers for TolDCs identification, developing shorter DCs differentiation methods and obtaining TolDCs with a stable phenotype. We describe here, a short-term protocol for TolDCs generation, which are characterized in terms of phenotypic markers, cytokines secretion profile, CD4+ T cell-stimulatory ability and migratory capacity.
  
  
  METHODS
  TolDCs from healthy donors were generated by modulation with dexamethasone plus monophosphoryl lipid A (MPLA-tDCs). We performed an analysis of MPLA-tDCs in terms of yield, viability, morphology, phenotypic markers, cytokines secretion profile, stability, allogeneic and antigen-specific CD4+ T-cell stimulatory ability and migration capacity.
  
  
  RESULTS
  After a 5-day culture, MPLA-tDCs displayed reduced expression of costimulatory and maturation molecules together to an anti-inflammatory cytokines secretion profile, being able to maintain these tolerogenic features even after the engagement of CD40 by its cognate ligand. In addition, MPLA-tDCs exhibited reduced capabilities to stimulate allogeneic and antigen-specific CD4+ T cell proliferation, and induced an anti-inflammatory cytokine secretion pattern. Among potential tolerogenic markers studied, only TLR-2 was highly expressed in MPLA-tDCs when compared to mature and immature DCs. Remarkable, like mature DCs, MPLA-tDCs displayed a high CCR7 and CXCR4 expression, both chemokine receptors involved in migration to secondary lymphoid organs, and even more, in an in vitro assay they exhibited a high migration response towards CCL19 and CXCL12.
  
  
  CONCLUSIONS
  We describe a short-term protocol for TolDC generation, which confers them a stable phenotype and migratory capacity to lymphoid chemokines, essential features for TolDCs to be used as therapeutics for autoimmunity and prevention of graft rejection.
authors.affiliation: Immune Regulation and Tolerance Research Group, Programa Disciplinario de Inmunología, ICBM, Facultad de Medicina, Universidad de Chile, Santiago, Chile.
authors.forename: Paulina
authors.initials: P
authors.lastname: García-González
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Morales
authors.forename: Lorena
authors.initials: L
authors.lastname: Hoyos
authors.forename: Jaxaira
authors.initials: J
authors.lastname: Maggi
authors.forename: Javier
authors.initials: J
authors.lastname: Campos
authors.forename: Bárbara
authors.initials: B
authors.lastname: Pesce
authors.forename: David
authors.initials: D
authors.lastname: Gárate
authors.forename: Milton
authors.initials: M
authors.lastname: Larrondo
authors.forename: Rodrigo
authors.initials: R
authors.lastname: González
authors.forename: Lilian
authors.initials: L
authors.lastname: Soto
authors.forename: Verónica
authors.initials: V
authors.lastname: Ramos
authors.forename: Pía
authors.initials: P
authors.lastname: Tobar
authors.forename: María Carmen
authors.initials: MC
authors.lastname: Molina
authors.forename: Karina
authors.initials: K
authors.lastname: Pino-Lagos
authors.forename: Diego
authors.initials: D
authors.lastname: Catalán
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Aguillón
mesh_terms: D015551:Autoimmunity; D015415:Biomarkers / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology; D002454:Cell Differentiation; D002465:Cell Movement; D018925:Chemokines / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects*; D003907:Dexamethasone / Q000494:pharmacology*; D005434:Flow Cytometry; D006801:Humans; D008050:Lipid A / Q000031:analogs & derivatives* / Q000494:pharmacology; D010641:Phenotype; D054400:Receptors, CCR7 / Q000378:metabolism; D019718:Receptors, CXCR4 / Q000378:metabolism
chemical_list: D015415:Biomarkers; C515370:CCR7 protein, human; C527276:CXCR4 protein, human; D018925:Chemokines; D016207:Cytokines; D008050:Lipid A; D054400:Receptors, CCR7; D019718:Receptors, CXCR4; D003907:Dexamethasone; C048436:monophosphoryl lipid A