Dendritic Cells Genetically Engineered to Express Fas Ligand Induce Donor-Specific Hyporesponsiveness and Prolong Allograft Survival

Abstract

Polarization of an immune response toward tolerance or immunity is dictated by the interactions between T cells and dendritic cells (DC), which in turn are modulated by the expression of distinct cell surface molecules, and the cytokine milieu in which these interactions are taking place. Genetic modification of DC with genes coding for specific immunoregulatory cell surface molecules and cytokines offers the potential of inhibiting immune responses by selectively targeting Ag-specific T cells. In this study, the immunomodulatory effects of transfecting murine bone marrow-derived DC with Fas ligand (FasL) were investigated. In this study, we show that FasL transfection of DC markedly augmented their capacity to induce apoptosis of Fas+ cells. FasL-transfected DC inhibited allogeneic MLR in vitro, and induced hyporesponsiveness to alloantigen in vivo. The induction of hyporesponsiveness was Ag specific and was dependent on the interaction between FasL on DC and Fas on T cells. Finally, we show that transfusion of FasL-DC significantly prolonged the survival of fully MHC-mismatched vascularized cardiac allografts. Our findings suggest that DC transduced with FasL may facilitate the development of Ag-specific unresponsiveness for the prevention of organ rejection. Moreover, they highlight the potential of genetically engineering DC to express other genes that affect immune responses.

Human myeloid dendritic cells transduced with an adenoviral interleukin-10 gene construct inhibit human skin graft rejection in humanized NOD-scid chimeric mice

“…[30][31][32] Recently, DC have also been modified with Fas ligand and proven capable of prolonging allograft survival in a cardiac transplantation model. 33 It is clear from other studies that the form of IL-10 is important to its biological effect. Mammalian or cellular IL-10 may potentially be immunostimulatory.…”

Mature But Not Immature Fas Ligand (CD95L)-Transduced Human Monocyte-Derived Dendritic Cells Are Protected from Fas-Mediated Apoptosis and Can Be Used as Killer APC

“…However, all these studies were performed using murine DC, B cells, macrophages, or cell lines as APC (12)(13)(14)(15)(16)(17)(18)(19)(20)(21)(22)(23)39). Therefore, for future clinical application, it had to be determined whether human DC can be efficiently transduced with FasL and whether these cells would be capable of eliminating Fas ϩ target cells.…”

Immune Modulation and Tolerance Induction by RelB-Silenced Dendritic Cells through RNA Interference

“…DC were generated from bone marrow progenitor cells, as previously described (24). Briefly, bone marrow cells were flushed from the femurs and tibias of C57BL/6 mice (The Jackson Laboratory), and then washed and cultured in 24-well plates (2 ϫ 10 6 cells/ml) in 2 ml of RPMI 1640 complete medium supplemented with 2 mM L-glutamine, 100 U/ml penicillin, 100 g of streptomycin, 50 M 2-ME, and 10% FCS (all from Invitrogen Life Technologies), supplemented with recombinant GM-CSF (10 ng/ml; PeproTech) and recombinant mouse IL-4 (10 ng/ml; PeproTech).…”