TY - JOUR
T1 - A short protocol using dexamethasone and monophosphoryl lipid A generates tolerogenic dendritic cells that display a potent migratory capacity to lymphoid chemokines
AU - García-González, Paulina
AU - Morales, Rodrigo
AU - Hoyos, Lorena
AU - Maggi, Jaxaira
AU - Campos, Javier
AU - Pesce, Bárbara
AU - Gárate, David
AU - Larrondo, Milton
AU - González, Rodrigo
AU - Soto, Lilian
AU - Ramos, Verónica
AU - Tobar, Pía
AU - Molina, María Carmen
AU - Pino-Lagos, Karina
AU - Catalán, Diego
AU - Aguillón, Juan Carlos
N1 - Funding Information:
The authors thank to Dr. Catharien Hilkens for helping us in the standardization of the in vitro migration assay, Mr. Alvaro Rojas for flow cytometry acquisition and Mrs. Juana Orellana, Mrs. Nancy Fabres and Mrs. Ruth Mora for technical assistance. Supported by Fondecyt-Chile 110–0102, Millennium Institute on Immunology and Immunotherapy-P09-016-F and Fundación Ciencia Translacional from Chile.
PY - 2013/5/24
Y1 - 2013/5/24
N2 - 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.Conclusion: 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.
AB - 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.Conclusion: 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.
KW - CCR7
KW - CXCR4
KW - Cell-based therapy
KW - Chemokine receptors
KW - Monocyte-derived dendritic cells
KW - Tolerance
UR - http://www.scopus.com/inward/record.url?scp=84878005244&partnerID=8YFLogxK
U2 - 10.1186/1479-5876-11-128
DO - 10.1186/1479-5876-11-128
M3 - Article
C2 - 23706017
AN - SCOPUS:84878005244
SN - 1479-5876
VL - 11
JO - Journal of Translational Medicine
JF - Journal of Translational Medicine
IS - 1
M1 - 128
ER -