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Comparative study of clinical grade human tolerogenic dendritic cells

BACKGROUND: The 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 appl...

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Autores principales: Naranjo-Gómez, M, Raïch-Regué, D, Oñate, C, Grau-López, L, Ramo-Tello, C, Pujol-Borrell, R, Martínez-Cáceres, E, Borràs, Francesc E
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3141500/
https://www.ncbi.nlm.nih.gov/pubmed/21658226
http://dx.doi.org/10.1186/1479-5876-9-89
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author Naranjo-Gómez, M
Raïch-Regué, D
Oñate, C
Grau-López, L
Ramo-Tello, C
Pujol-Borrell, R
Martínez-Cáceres, E
Borràs, Francesc E
author_facet Naranjo-Gómez, M
Raïch-Regué, D
Oñate, C
Grau-López, L
Ramo-Tello, C
Pujol-Borrell, R
Martínez-Cáceres, E
Borràs, Francesc E
author_sort Naranjo-Gómez, M
collection PubMed
description BACKGROUND: The 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. METHODS: Tolerogenic 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. RESULTS: Differences 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. CONCLUSIONS: Our 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.
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spelling pubmed-31415002011-07-23 Comparative study of clinical grade human tolerogenic dendritic cells Naranjo-Gómez, M Raïch-Regué, D Oñate, C Grau-López, L Ramo-Tello, C Pujol-Borrell, R Martínez-Cáceres, E Borràs, Francesc E J Transl Med Research BACKGROUND: The 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. METHODS: Tolerogenic 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. RESULTS: Differences 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. CONCLUSIONS: Our 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. BioMed Central 2011-06-09 /pmc/articles/PMC3141500/ /pubmed/21658226 http://dx.doi.org/10.1186/1479-5876-9-89 Text en Copyright ©2011 Naranjo-Gómez et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Naranjo-Gómez, M
Raïch-Regué, D
Oñate, C
Grau-López, L
Ramo-Tello, C
Pujol-Borrell, R
Martínez-Cáceres, E
Borràs, Francesc E
Comparative study of clinical grade human tolerogenic dendritic cells
title Comparative study of clinical grade human tolerogenic dendritic cells
title_full Comparative study of clinical grade human tolerogenic dendritic cells
title_fullStr Comparative study of clinical grade human tolerogenic dendritic cells
title_full_unstemmed Comparative study of clinical grade human tolerogenic dendritic cells
title_short Comparative study of clinical grade human tolerogenic dendritic cells
title_sort comparative study of clinical grade human tolerogenic dendritic cells
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3141500/
https://www.ncbi.nlm.nih.gov/pubmed/21658226
http://dx.doi.org/10.1186/1479-5876-9-89
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