IL4 and IFNalpha generation of dendritic cells reveals great migratory potential and NFkB and cJun expression in IL4DCs

Abstract

Dendritic cells (DCs) recently revealed as a potent tumor vaccine component, are commonly differentiated from monocytes by cultivation with IL-4 and GM-CSF. Despite the different opinions, the use of IFNalpha can promote DCs differentiation and activation. The aim of this study was to compare the functionality and phenotypic characterization of monocyte-derived DC generated by IL-4 (IL4DC) and IFNalpha (IFNalphaDC) modified protocols. To this aim, we investigated the expression of maturation markers, co-stimulatory molecules, relevant miRNA, cytokine and migratory profiles and the functional ability of these cells to stimulate autologous T cells in vitro. We herein investigated the molecular mechanism underlying the parameters previously described, as the relative expression of NF-kB p65, c-fos and c-jun, transcription factors. Our results demonstrated that IL4DC presented a stable phenotype, an increase in migratory capacity and NF-KB activation, in addition to lower levels of miR-146 a and miR-221. We believe that the IL4DC migratory potential and increase in NFkBp65 expression may be involved in higher IL12 expression and migration, suggesting a preferential activation of TH1 immune responses by IL4DC.

miR-146 and miR-155: Two Key Modulators of Immune Response and Tumor Development

“…An initial study showed that miR-146 expression was lower in dendritic cells originated from monocytes grown in the presence of IL-4 and GM-CSF, compared to dendritic cells originated by cultivation of monocytes in the presence of IFN-α [ 18 ]. The lower miR-146 expression in these cells is accompanied by an increased migratory capacity and NF-ĸB expression [ 19 ]. miR-146a and particularly miR-146b expression were markedly up-modulated during maturation of monocytes to dendritic cells using GM-CSF and IL-4: particularly, miR-146a and miR-146b expression was about 10–40-fold, respectively, higher than in monocytes [ 20 ].…”

“…pDCs constitutively express miR-146a levels much higher than mature monocytes: however, miR-146a expression is strongly activated in monocytic cells, but not in pDCs by IFN-γ, IL-1β and lipopolysaccharide (LPS) [ 22 ]. miR-146a expression is up-modulated during differentiation of pDCs by the transcription factor PU.1, and by itself, miR-146a was unable to affect DC differentiation [ 19 ]. miR-146a expression in pDCs is markedly up-modulated upon activation of primary pDCs using TLR7 or TLR9 agonists [ 23 ].…”

Klotho sensitive regulation of dendritic cell functions by vitamin E

“…Accordingly, the LPS-induced increases in expression of IL-12p70 protein in, migration of and ROS production in VitE-treated DCs were significantly reduced, all effects were virtually abolished after suppressing klotho synthesis with klotho siRNA. Moreover, the effects are modulated through NF-κB activation [15, 31]. Therefore, this signaling could participate in regulation of klotho expression.…”

Small Particles, Big Effects: The Interplay Between Exosomes and Dendritic Cells in Antitumor Immunity and Immunotherapy

“…The development and enhancement of these cell culture techniques led to increasing cell yields throughout the 1990s, which allowed the exploration of the pulse of DCs with tumor antigens as a new potential anticancer vaccines [59,60]. The most commonly used conventional protocols employ peripheral blood mononuclear leukocytes that are differentiated ex vivo into monocytes and later on into mature dendritic cells [61,62], which are then pulsed with specific tumor antigens in various forms, such as synthetic peptides [63], tumor nucleic acid [64], and apoptotic bodies [65] among others.…”