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Journal of Translational Medicine

, 9:89

First Online: 09 June 2011Received: 18 January 2011Accepted: 09 June 2011DOI: 10.1186-1479-5876-9-89

Cite this article as: Naranjo-Gómez, M., Raïch-Regué, D., Oñate, C. et al. J Transl Med 2011 9: 89. doi:10.1186-1479-5876-9-89

Abstract

BackgroundThe use of tolerogenic DCs is a promising therapeutic strategy for transplantation and autoimmune disorders. Immunomodulatory DCs are primarily generated from monocytes MDDCs for in vitro experiments following protocols that fail to fulfil the strict regulatory rules of clinically applicable products. Here, we compared the efficacy of three different tolerance-inducing agents, dexamethasone, rapamycin and vitamin D3, on DC biology using GMP Good Manufacturing Practice or clinical grade reagents with the aim of defining their use for human cell therapy.

MethodsTolerogenic MDDCs were generated by adding tolerogenic agents prior to the induction of maturation using TNF-α, IL-β and PGE2. We evaluated the effects of each agent on viability, efficiency of differentiation, phenotype, cytokine secretion and stability, the stimulatory capacity of tol-DCs and the T-cell profiles induced.

ResultsDifferences relevant to therapeutic applicability were observed with the cellular products that were obtained. VitD3-induced tol-DCs exhibited a slightly reduced viability and yield compared to Dexa-and Rapa-tol-DCs. Phenotypically, while Dexa-and VitD3-tol-DCs were similar to immature DCs, Rapa-tol-DCs were not distinguishable from mature DCs. In addition, only Dexa-and moderately VitD3-tol-DCs exhibited IL-10 production. Interestingly, in all cases, the cytokine secretion profiles of tol-DCs were not modified by a subsequent TLR stimulation with LPS, indicating that all products had stable phenotypes. Functionally, clearly reduced alloantigen T cell proliferation was induced by tol-DCs obtained using any of these agent. Also, total interferon-gamma IFN-γ secretion by T cells stimulated with allogeneic tol-DCs was reduced in all three cases, but only T cells co-cultured with Rapa-tol-DCs showed impaired intracellular IFN-γ production. In addition, Rapa-DCs promoted CD4+ CD127 low-negative CD25high and Foxp3+ T cells.

ConclusionsOur results demonstrate contrasting influences of different clinical-grade pharmacological agents on human tol-DC generation. This should be taken into account for decisions on the use of a specific agent for the appropriate cellular therapy in the context of a particular disease.

List of abbreviationsDCdendritic cell

Dexadexamethasone

GMPGood Manufacturing Practice

IFN-γInterferon-gamma

Ioionomycin

MDDCMonocyte Derived DC

PBMCsPeripheral Blood Mononuclear Cells

PMAphorbol 12-myristate 13-acetate

Raparapamycin

tol-DCtolerogenic DCs

Tregsregulatory T cells

VitD3vitamin D3.

Electronic supplementary materialThe online version of this article doi:10.1186-1479-5876-9-89 contains supplementary material, which is available to authorized users.

E Martínez-Cáceres and Francesc E Borràs contributed equally to this work.

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Author: M Naranjo-Gómez - D Raïch-Regué - C Oñate - L Grau-López - C Ramo-Tello - R Pujol-Borrell - E Martínez-Cáceres - Fra

Source: https://link.springer.com/



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