Allostimulatory Effects of Dendritic Cells with Characteristic Features of a Regulatory Phenotype.

Introduction: Tolerogenic dendritic cells (DCs) have the potential to prolong graft survival after transplantation. Tolerogenic DCs are in general characterized by a low expression of co-stimulatory molecule and a high IL-10:IL-12 production ratio. Based on promising results with earlier used alternatively activated DCs, we aimed to generate in culture potentially tolerogenic DC by simultaneously blocking GSK3 by lithium chloride (LiCl) and stimulating TLR2 by PAM₃CysSerLys₄. Materials and Methods: Bone marrow-derived LiClPAM₃ DCs were generated by the addition of LiCl 24 hours before harvesting, and one hour later PAM₃CysSerLys₄. The phenotype of the DCs was assessed by determining the expression of co-stimulatory molecules in flow cytometry and cytokine production in ELISA, whereas their functional properties were tested in a mixed lymphocyte reaction. A fully MHC mismatched heterotopic heart transplant preceded by infusion of donor-derived LiClPAM₃ DC was performed to assess the tolerogenic potential of LiClPAM₃ DCs in vivo. Results: LiClPAM₃ DCs displayed a tolerogenic phenotype accompanied with a low expression of co-stimulatory molecules and a high IL-10:IL-12 production ratio. However, in mixed lymphocyte reaction, LiClPAM₃ DCs appeared superior in T cell stimulation, and induced Th1 and Th17 differentiation. Moreover, mice pretreated with LiClPAM₃ DC displayed a reduced graft survival. Analysis of LiClPAM₃ DC culture supernatant revealed high levels of CXCL-1, which was also found in supernatants of co-cultures of LiClPAM₃ DC and T cells. Nevertheless, we could not show a role for CXCL-1 in T cell proliferation or activation in vitro. Discussion: LiClPAM₃ DCs display in vitro a tolerogenic phenotype with a high IL-10:IL-12 ratio, but appeared to be highly immunogenic, since allograft rejection was accelerated. As yet unidentified LiClPAM₃ DC-derived factors, may explain the immunogenic character of LiClPAM₃ DCs in vivo. [ABSTRACT FROM AUTHOR]