Purpose Dexamethasone (Dex) has long been used as a potent immunosuppressive agent in the treatment of inflammatory and autoimmune diseases, despite serious side effects. In the present study, Dex and model antigen ovalbumin (OVA) were encapsulated with poly(lactic-co-glycolic acid) to deliver Dex and OVA preferentially to phagocytic cells, reducing systemic side effects of Dex. The OVA-specific immune tolerance-inducing activity of the nanoparticles (NPs) was examined. Methods Polymeric NPs containing OVA and Dex (NP[OVA+Dex]) were prepared by the water-in-oil-in-water double emulsion solvent evaporation method. The effects of NP[OVA+Dex] on the maturation and function of immature dendritic cells (DCs) were examined in vitro. Furthermore, the OVA-specific immune tolerizing effects of NP[OVA+Dex] were confirmed in mice that were intravenously injected or orally fed with the NPs. Results Immature DCs treated in vitro with NP[OVA+Dex] did not mature into immunogenic DCs but instead were converted into tolerogenic DCs. Furthermore, profoundly suppressed generation of OVA-specific cytotoxic T cells and production of OVA-specific IgG were observed in mice injected with NP[OVA+Dex], whereas regulatory T cells were concomitantly increased. Feeding of mice with NP[OVA+Dex] also induced OVA-specific immune tolerance. Conclusion The present study demonstrates that oral feeding as well as intravenous injection of poly(lactic-co-glycolic acid) NPs encapsulating both antigen and Dex is a useful means of inducing antigen-specific immune tolerance, which is crucial for the treatment of autoimmune diseases.

The active form of vitamin D 3 , 1,25-dihydroxyvitamin D 3 (aVD 3 ), is known to exert beneficial effects in the treatment of autoimmune diseases because of its immunosuppressive effects. However, clinical application of aVD 3 remains limited because of the potential side effects, particularly hypercalcemia. Encapsulation of aVD 3 within biodegradable nanoparticles (NPs) would enhance the delivery of aVD 3 to antigen presenting cells, while preventing the potential systemic side effects of aVD 3 . In the present study, polymeric NPs containing ovalbumin (OVA) and aVD 3 (NP[OVA+aVD 3 ]) were prepared via the water-in-oil-in-water double emulsion solvent evaporation method, after which their immunomodulatory effects were examined. Bone marrow-derived immature dendritic cells (DCs) treated with NP(OVA+aVD 3 ) did not mature into immunogenic DCs but were converted into tolerogenic DCs, which express low levels of co-stimulatory molecules and MHC class II molecules, produce lower levels of pro-inflammatory cytokines while increasing the production of IL-10 and TGF-β, and induce the generation of Tregs. Intravenous injection with NP(OVA+aVD 3 ) markedly suppressed the generation of OVA-specific CTLs in mice. Furthermore, OVA-specific immune tolerance was induced in mice orally administered with NP(OVA+aVD 3 ). These results show that biodegradable NPs encapsulating both antigen and aVD 3 can effectively induce antigen-specific immune suppression.

No abstract

Thapsigargin (TGN) is a potent and selective inhibitor of sarco-endoplasmic Ca2+-ATPase, leading to rapid elevation of cytoplasmic Ca2+ concentration. Previous reports have shown that TGN increases the production of various cytokines from macrophages and dendritic cells. Here, we examine the effects of TGN on murine T cells. Nanomolar concentrations of TGN are a significant inducer of IL-2 production with full activity at 50 nM. Micromolar concentrations of TGN, however, are inhibitory to IL-2 production and T cell proliferation. The IL-2 production-inducing activity of TGN is much more prominent when T cells are primed with concanavalin A or anti-CD3 mAb, and is due to the increase of cytoplasmic Ca2+ concentration. TGN at 50 nM does not affect interferon-gamma or IL-4 production from T cells. Thus, the present study shows that low nanomolar concentrations of TGN could be useful in potentiating IL-2 production from antigen-primed T cells.

Dexamethasone (Dex) is known to reduce inflammation and autoimmunity by modulating dendritic cell (DC) function, maturation, and differentiation. Nanoparticles (NPs) prepared using biodegradable and biocompatible poly(lactic-co-glycolic acid) (PLGA) can be used to target-delivery vehicles to DCs. In the present study, PLGA nanoparticles containing ovalbumin (OVA) and Dex (NP[OVA+Dex]) were prepared by W1/O/W2 double emulsion solvent evaporation method, and evaluated their antigen-specific immune tolerizing effects. DCs treated with NP[OVA+Dex] were decreased in the expression of MHC class II (I-Ab) and co-stimulatory molecules (CD80 and CD86), and increased in the proportion of CD86−PD-L1+ cell population. DCs treated with NP[OVA+Dex] were reduced in the production of pro-inflammatory cytokine, such as IL-1β, IL-6, IL-12p40 and TNF-α, whereas the production of immunosurpressive cytokine IL-10 was enhanced. Treatment of DCs with NP[OVA+Dex] were diminished in MHC class II-restricted OVA presentation and allogenic T cell stimulatory capacity. NP[OVA+Dex]-treated DCs induced CD4+CD25+FoxP3+ regulatory T cell differentiation from CD4+CD25− T cells isolated from normal, or OT-II mice. Mice injected i.v., or i.g. with NP[OVA+Dex] were suppressed in both inducing OVA-specific CTL responses and producing OVA-specific IgG. The present study demonstrates that NPs encapsulated with antigen and Dex are useful tools in inducing antigen-specific immune tolerance.