Antisense oligonucleotides down-regulating costimulation confer diabetes-preventive properties to nonobese diabetic mouse dendritic cells

Diabetes Institute, Department of Pediatrics, University of Pittsburgh School of Medicine, PA 15213, USA.
The Journal of Immunology (Impact Factor: 5.36). 11/2004; 173(7):4331-41. DOI: 10.4049/jimmunol.173.7.4331
Source: PubMed

ABSTRACT Phenotypically "immature" dendritic cells (DCs), defined by low cell surface CD40, CD80, and CD86 can elicit host immune suppression in allotransplantation and autoimmunity. Herein, we report the most direct means of achieving phenotypic immaturity in NOD bone marrow-derived DCs aiming at preventing diabetes in syngeneic recipients. CD40, CD80, and CD86 cell surface molecules were specifically down-regulated by treating NOD DCs ex vivo with a mixture of antisense oligonucleotides targeting the CD40, CD80, and CD86 primary transcripts. The incidence of diabetes was significantly delayed by a single injection of the engineered NOD DCs into syngeneic recipients. Insulitis was absent in diabetes-free recipients and their splenic T cells proliferated in response to alloantigen. Engineered DC promoted an increased prevalence of CD4(+)CD25(+) T cells in NOD recipients at all ages examined and diabetes-free recipients exhibited significantly greater numbers of CD4(+)CD25(+) T cells compared with untreated NOD mice. In NOD-scid recipients, antisense-treated NOD DC promoted an increased prevalence of these putative regulatory T cells. Collectively, these data demonstrate that direct interference of cell surface expression of the major costimulatory molecules at the transcriptional level confers diabetes protection by promoting, in part, the proliferation and/or survival of regulatory T cells. This approach is a useful tool by which DC-mediated activation of regulatory T cells can be studied as well as a potential therapeutic option for type 1 diabetes.

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    • "Three patients received control DC generated in the presence of GM- CSF and IL-4 and seven patients received immunosuppressive DC generated in the presence of GM-CSF, IL-4, and antisense oligonucleotides targeting CD40, CD80, and CD86 transcripts. Use of tolerogenic DC generated with these antisense oligonucleotides was shown previously by the same team to have a preventive and curative effect on diabetes in NOD mice (Machen et al., 2004). This Phase I study demonstrated that intradermal injections of autologous TolDC (both control and immunosuppressive DC) are well-tolerated and safe in diabetic patients; no adverse effects or toxicity was observed. "
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    ABSTRACT: The use of immunosuppressive (IS) drugs to treat transplant recipients has markedly reduced the incidence of acute rejection and early graft loss. However, such treatments have numerous adverse side effects and fail to prevent chronic allograft dysfunction. In this context, therapies based on the adoptive transfer of regulatory cells are promising strategies to induce indefinite transplant survival. The use of tolerogenic dendritic cells (DC) has shown great potential, as preliminary experiments in rodents have demonstrated that administration of tolerogenic DC prolongs graft survival. Recipient DC, Donor DC, or Donor Ag-pulsed recipient DC have been used in preclinical studies and administration of these cells with suboptimal immunosuppression increases their tolerogenic potential. We have demonstrated that autologous unpulsed tolerogenic DC injected in the presence of suboptimal immunosuppression are able to induce Ag-specific allograft tolerance. We derived similar tolerogenic DC in different animal models (mice and non-human primates) and confirmed their protective abilities in vitro and in vivo. The mechanisms involved in the tolerance induced by autologous tolerogenic DC were also investigated. With the aim of using autologous DC in kidney transplant patients, we have developed and characterized tolerogenic monocyte-derived DC in humans. In this review, we will discuss the preclinical studies and describe our recent results from the generation and characterization of tolerogenic monocyte-derived DC in humans for a clinical application. We will also discuss the limits and difficulties in translating preclinical experiments to theclinic.
    Frontiers in Immunology 08/2012; 3:218. DOI:10.3389/fimmu.2012.00218
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    • "One such example is the use of immunoregulatory dendritic cells (iDC), autologous monocytederived DCs in which the CD40, CD80 and CD86 co-stimulatory molecules are downregulated by antisense molecules before reintroduction . An approved clinical safety study (NCT00445913) will evaluate the success achieved in NOD mice [40] [41]. Second, from a clinical standpoint it will prove useful to pre-screen patients for a specific type of treatment based on the presence or absence of certain 'biomarkers', for example autoreactive T cells. "
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    ABSTRACT: Redirection of immune responses by manipulation of antigen-presenting cells is an emerging strategy for immunosuppressive treatment of autoimmune diseases. In vivo expansion of dendritic cells (DC) by Fms-like tyrosine kinase-3 (Flt3)-Ligand (FL) treatment was shown to delay diabetes onset in the NOD model of autoimmune diabetes. However, we show here that Flt3 stimulation actually accelerates autoimmunity when autoreactive CD8 T cells are detectable in blood prior to treatment. With autoreactive CD8 cells present, the capacity of FL to expand DCs and induce Treg remained intact, but both numbers and the functional response of islet-specific CD8s were boosted. Also, the inhibitory receptor PD-1 on (autoreactive) CD8 T cells and its ligand PD-L1 on Treg were no longer upregulated. These data highlight the need to pre-screen for T cell autoreactivity prior to generalized DC expansion and illustrate how accelerated disease can occur when the intended initiation of regulatory mechanisms is impaired later in diabetogenesis.
    Journal of Autoimmunity 06/2010; 34(4-34):445-452. DOI:10.1016/j.jaut.2009.11.010
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