Vaccine 30 (2012) 378–387 Contents lists available at SciVerse ScienceDirect Vaccine j ourna l ho me pag e: www.elsevier.com/locate/vaccine Differential effects of monophosphoryl lipid A and cytokine cocktail as maturation stimuli of immunogenic and tolerogenic dendritic cells for immunotherapy Dàlia Raïch-Regué a , Mar Naranjo-Gómez a , Laia Grau-López b , Cristina Ramo b , Ricardo Pujol-Borrell a , Eva Martínez-Cáceres a,∗,1 , Francesc E. Borràs a,∗,1 a Laboratory of Immunobiology for Research and Diagnosis (LIRAD), Blood and Tissue Bank (BST), Dept. of Cell Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, Institut Investigació Germans Trias i Pujol (IGTP), Badalona, Barcelona, Spain b Multiple Sclerosis Unit, Dept. of Neurosciences, Hospital Universitari Germans Trias i Pujol, Dept. of Medicine, Universitat Autònoma de Barcelona, Barcelona, Spain a r t i c l e i n f o Article history: Received 18 April 2011 Received in revised form 10 August 2011 Accepted 28 October 2011 Available online 12 November 2011 Keywords: Human dendritic cells Maturation stimuli Clinical-grade TLR-agonist DC-based vaccine Cell therapy Interleukin-17 a b s t r a c t Immunotherapy using monocyte-derived dendritic cells (MDDC) is increasingly being considered as alter- native therapeutic approach in cancer, infectious diseases and also in autoimmunity when patients are not responsive to conventional treatments. In general, generation of MDDC from monocytes is induced in the presence of GM-CSF and IL-4, and a maturation stimulus is added to the culture to obtain mature DCs suitable for therapy. For DC maturation, different combinations of pro-inflammatory mediators and Toll-like receptor ligands have been tested, obtaining DCs that differ in their properties and the type of immune response they promote. Therefore, it is necessary to find an optimal cytokine environment for DC maturation to obtain a cellular product suitable for DC-based immunotherapeutic protocols. In this study, we have evaluated in vitro the effects of different maturation stimuli on the viability, phenotype, cytokine profile, stability and functionality of immunogenic and tolerogenic (1,25- dihydroxyvitamin D 3 -treated) MDDC. Maturation was induced using the clinical grade TLR4-agonist: monophosphoryl lipid A (LA), compared to the traditional cytokine cocktail (CC; clinical grade TNF-, IL-1, PGE2) and a combination of both. Our results showed the combination of CC + LA rendered a potent immunogenic DC population that induced the production of IFN- and IL-17 in allogeneic co-cultures, suggesting a Th17 polarization. Moreover, these immunogenic DCs showed a high surface expression of CD83, CD86, HLA-DR and secre- tion of IL-12p70. When aiming to induce tolerance, using LA to generate mature TolDC did not represent a clear advantage, and the stability and the suppressive capability exhibited by CC-matured TolDC may rep- resent the best option. Altogether, these findings demonstrate the relevance of an appropriate maturation stimulus to rationally modulate the therapeutic potential of DCs in immunotherapy. © 2011 Elsevier Ltd. All rights reserved. 1. Introduction Since dendritic cells (DCs) were identified as master cells in the modulation of the immune response, a lot of effort has been made to bring their potential to the clinical setting [1–3]. The definition of culture methods to derive DCs from monocytes (MDDC) [4,5] Abbreviations: MDDC, monocyte-derived dendritic cells; DCs, dendritic cells; TolDC, tolerogenic DCs; MatDC, immunogenic DCs; TLR, toll-like receptor; vitD3, 1,25-dihydroxyvitamin D3; LA, monophosporyl lipid A; LPS, lipopolysaccharide; CC, cytokine cocktail; GMP, good manufacturing practices; IFN-, interferon gamma. ∗ Corresponding authors at: LIRAD-BST, Institut d’Investigació Germans Trias i Pujol, Ctra. del Canyet s/n, Edifici “Escoles”, 08916 Badalona (Barcelona), Spain. Tel.: +34 93 497 86 66 8671; fax: +34 93 497 86 68. E-mail address: feborras@igtp.cat (F.E. Borràs). 1 Shared senior co-authorship. established the starting point to the development of new poten- tial therapeutic approaches for several diseases that are refractory to conventional treatments. Among those, cancer and infectious diseases were first treated with MDDC, reporting partially satis- factory results [1,6–8]. The extensive knowledge generated in the past has even increased the interest in considering immunother- apy using MDDC as an alternative therapeutic approach in patients non-responding to current treatments in cancer, infectious diseases and lately, in autoimmune disorders. Initially, MDDC are obtained using similar procedures, but each target disease must be particularly envisaged, as the final desired effect would be different. To date, most clinical studies have used a combination of pro-inflammatory cytokines to obtain the so-called “mature dendritic cells” in vitro from monocyte precursors [9]. The standard cytokine cocktail (CC) containing TNF-, IL-1, IL-6 and PGE-2, has largely proved to effectively mature DCs in vitro [10]. 0264-410X/$ – see front matter © 2011 Elsevier Ltd. All rights reserved. doi:10.1016/j.vaccine.2011.10.081