Transfection of Vitamin D3-Induced Tolerogenic Dendritic Cells for the Silencing of Potential Tolerogenic Genes. Identification of CSF1R-CSF1 Signaling as a Glycolytic Regulator.

María José Mansilla,Eva Martínez-Cáceres,Bibiana Quirant-Sánchez,Ares Sellés-Rius,Neus Figa-Martín,Jaume Barallat,Aina Teniente-Serra,Federico Fondelli,Iñigo González-Larreategui

doi:10.3390/ijms22147363

Abstract

The use of autologous tolerogenic dendritic cells (tolDC) has become a promising strategy to re-establish immune tolerance in autoimmune diseases. Among the different strategies available, the use of vitamin D3 for the generation of tolDC (VitD3-tolDC) has been widely tested because of their immune regulatory properties. To identify molecules and pathways involved in the generation of VitD3-tolDC, we established an easy and fast gene silencing method based on the use of Viromer blue to introduce siRNA into monocytes on day 1 of culture differentiation. The analysis of the effect of CD209 (DC-SIGN) and CD115 (CSF1R) down-modulation on the phenotype and functionality of transfected VitD3-tolDC revealed a partial role of CD115 in their tolerogenicity. Further investigations showed that CSF1R-CSF1 signaling is involved in the induction of cell metabolic reprogramming, triggering glycolysis to produce high amounts of lactate, a novel suppressive mechanism of T cell proliferation, recently found in autologous tolerogenic dendritic cells (ATDCs).

Highlights

Since the beginning of the 21st century, researchers have focused their efforts on using dendritic cells (DC) as a powerful tool to control altered immune responses and restore the homeostatic balance
Through a microarray analysis we found upregulated in VitD3-tolerogenic dendritic cells (tolDC), compared to mature DC (mDC), the genes CD115 as well as CSF1, among others [17]
DC-SIGN expression is induced by CSF1 + IL-4 and is required for the development of the above-mentioned suppressive capacity of macrophages through IL-10 production [19,24]. All these findings suggest a possible role of CD115 and CD209 molecules in the tolerogenic potential of VitD3-tolDC

Summary

Introduction

Since the beginning of the 21st century, researchers have focused their efforts on using dendritic cells (DC) as a powerful tool to control altered immune responses and restore the homeostatic balance. DC are able to control T cell activation, making them a promising treatment for cancer therapy, and for restoring immune tolerance in autoimmune diseases and avoiding allograft rejection [2,3] In this context, tolerogenic dendritic cells (tolDC) represent a recent emerging strategy to induce a specific and long-term re-establishment of the immune tolerance towards self-antigens causing the immunogenic response of autoimmune diseases such as multiple sclerosis (MS), rheumatoid arthritis (RA) or type-1 diabetes (T1D). Tolerogenic dendritic cells (tolDC) represent a recent emerging strategy to induce a specific and long-term re-establishment of the immune tolerance towards self-antigens causing the immunogenic response of autoimmune diseases such as multiple sclerosis (MS), rheumatoid arthritis (RA) or type-1 diabetes (T1D) In this way, a tolDC strategy avoids the chronic and general immunosuppression of current treatments producing different serious adverse effects in patients [4]. TolDC express low levels of co-stimulatory molecules (CD80, CD83, CD86)

Methods

Results

Conclusion