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: 4.92). 11/2004; 173(7):4331-41. DOI: 10.4049/jimmunol.173.7.4331
Source: PubMed


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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    • "In fact, we first demonstrated that autologous DC generated in the presence of antisense oligonucleotides (AS-ODN) targeting the CD40, CD80 and CD86 costimulation protein primary transcripts could prevent and reverse T1D in the NOD strain via upregulation of Foxp3+ Tregs and novel IL-10+ B-regulatory cells (Bregs) [54] [55] [56] [57] and that human embodiments of these cell products were safe and potentially of some benefit in humans, in a phase I clinical trial [58]. "
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    • "In addition to suppression of inflammatory cytokine synthesis, the activated DCs would provide an element of safety because they have a limited lifetime of approximately 5 days and therefore would have only a transient effect on the immune response [139]. Additional studies have demonstrated prevention and reversal of type 1 diabetes in NOD mice using costimulation impaired, immunosuppressive bone marrow-derived DCs generated ex vivo with a mixture of antisense oligonucleotides targeting the primary transcripts of DC costimulatory factors CD40, CD80, and CD86 [140]. Phase 1 clinical trials show that the vaccine is well tolerated in patients [141]. "
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    • "We now add suppressive B-cells/Bregs to the list. RA production by iDC can explain, in part, our observations of increased CD4+ CD25+ T-cell frequency in successfully-treated NOD mice [23] as well as the increased frequency of suppressive B-cells, which express, as we have shown herein, RA receptors. The seminal findings of Clare-Salzer and colleagues [102], who showed that B-cells were increased in frequency in NOD mice injected DC from syngeneic pancreatic lymph nodes, could represent an a process of DC-induced suppressive B-cell expansion, as part of a therapeutic mechanism as they originally reported. "
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