A short protocol using dexamethasone and monophosphoryl lipid A generates tolerogenic dendritic cells that display a potent migratory capacity to lymphoid chemokines

Journal of Translational Medicine

Juan Carlos Aguillón¹  Diego Catalán¹  Karina Pino-Lagos¹  María Carmen Molina³  Pía Tobar⁴  Verónica Ramos¹  Lilian Soto⁵  Rodrigo González²  Milton Larrondo²  David Gárate⁴  Bárbara Pesce¹  Javier Campos¹  Jaxaira Maggi¹  Lorena Hoyos¹  Rodrigo Morales¹  Paulina García-González¹ 

[1] Millennium Institute on Immunology and Immunotherapy, Santiago, Chile;Banco de Sangre, Hospital Clínico de la Universidad de Chile, Santiago, Chile;Laboratorio de Evasión Inmune, Programa Disciplinario de Inmunología, ICBM, Facultad de Medicina, Universidad de Chile, Santiago, Chile;Immune Regulation and Tolerance Research Group, Programa Disciplinario de Inmunología, ICBM, Facultad de Medicina, Universidad de Chile, Santiago, Chile;Sección de Reumatología, Hospital Clínico de la Universidad de Chile, Santiago, Chile

关键词: Cell-based therapy;    CXCR4;    CCR7;    Chemokine receptors;    Monocyte-derived dendritic cells;    Tolerance;   

Others  :  827531
DOI  :  10.1186/1479-5876-11-128

 received in 2013-02-25, accepted in 2013-05-20,  发布年份 2013

【 摘 要 】

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.

【 授权许可】

   
2013 García-González et al.; licensee BioMed Central Ltd.

【 图 表 】

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