The modulation of mature dendritic cells from patients with type 1 diabetes using human periodontal ligament stem cells. An in-vitro study.

ABSTRACT

OBJECTIVE: This in vitro study aimed to investigate whether human periodontal ligament stem cells isolated from impacted third molars can modify the maturation and phenotype of monocyte-derived dendritic cells pulsed with GAD-65 obtained from patients with type 1 diabetes. BACKGROUND: Human periodontal ligament stem cells (PDLSCs) have been found to display cell surface marker characteristics similar to bone marrow stromal stem cells (BMSSCs). The immunosuppressive effects on dendritic cells (DCs), T and B cells as well as their low immunogenicity allow the use of PDLSCs in stem cell therapies for autoimmune diseases including type 1 diabetes (T1D). Studies on the immunomodulatory potential of PDLSCs in the context type 1 diabetes are lacking but are therefore worth pursuing. METHODS: CD14 + monocytes isolated from peripheral blood mononuclear cells (PBMNCs) of type 1 diabetic patients were differentiated into immature Dendritic Cells (iDCs) and then maturation was induced to generate Mature Dendritic Cells (mDCs). The mDCs were pulsed with human recombinant GAD-65 and then co-cultured with PDLSCs that were isolated from impacted third molars and characterized. The changes in the levels of differentiation and maturation surface markers on the dendritic cells were analyzed by flow cytometry at the immature state, mature state and after the co-culture experiment. The levels of the secreted cytokines; IL-6, IL-10, and TGF-ß were measured by ELISA in cell-free culture supernatant. RESULTS: PDLSCs exerted an immunosuppressive effect on fully mature dendritic cells from patients with type 1 diabetes. This immunoregulatory property of was apparent by the reduction of all maturation markers including CD80, CD83, CD86, CD40, CD1a, CD209 and HLA-DR. Moreover, there was a detection of high levels of anti-inflammatory cytokines in the co-culture supernatant media including a significant increase in the concentration of IL-6 and TGF-ß. CONCLUSIONS: The current in vitro study provides strong evidence that PDLSCs seem to be a very promising source for overcoming the autoimmune destruction seen in T1D as they exerted an immunosuppressive effect on monocyte derived mDCs from patients with T1D. Additional studies should be conducted to further reveal the immunomodulatory and suppressive properties of PDLSCs and their potential use in immunotherapy for this disease.