Autoantigen-specific protection of non-obese diabetic mice from cyclophosphamide-accelerated diabetes by vaccination with dendritic cells

Abstract

Aims/hypothesis. Dendritic cells (DCs) are professional antigen presenting cells involved in the initiation of primary immune responses and the preservation of peripheral tolerance. The aim of this study was to develop a DC vaccine for autoantigen-specific prevention of autoimmune diabetes. Methods. Splenocytes from diabetes-prone NOD mice were cultured in conditioned media to obtain a homogeneous DC sub-population for vaccination experiments. These cells were used to modulate autoimmune responses in NOD mice after synchronization of diabetes with cyclophosphamide. After immunisation with insulin-pulsed DCs the incidence of diabetes, the insulitis grade and the cytokine production was examined. Results. The long-term culture of splenocytes resulted in the generation of a cell line, termed NOD-DC1, which have a phenotype of myeloid DCs (CD11c, CD11b, DEC-205), express MHC class II and costimulatory molecules (CD40, CD80, CD86). The NOD-DC1 cells have preserved functional activity shown by the detection of a high antigen uptake capacity, the induction of a mixed lymphocyte reaction and stimuli-dependent IL-6 and TNF-α secretion. Vaccination with insulin-pulsed NOD-DC1 cells results in an antigen-specific prevention of diabetes. This was mediated by a reduction of the severity of insulitis and a decrease of T helper 1 effector cells. Conclusion/interpretation. We describe the generation of a DC line which confers protection from diabetes in an antigen-specific way. Our data shows that autoantigen-loaded DCs can induce strong immunoregulatory effects supporting the hypothesis that DCs are promising candidates to develop novel vaccines for the prevention of autoimmune diabetes. [Diabetologia (2003[Diabetologia ( ) 46:1357[Diabetologia ( -1365

1,25‐Dihydroxyvitamin D3 alters murine dendritic cell behaviour in vitro and in vivo

“…This may be attributed to the specific settings of allotransplantation with full MHC mismatch, where direct immune recognition will also be a major component, limiting the contribution of dendritic cells. The systemic in vivo use of 1,25(OH) 2 D 3 yields more pronounced results in animal models of autoimmunity [42,43], where dendritic cells play a more important role, as in the prevention of graft rejection. In the latter situation, combination with classical immunosuppressants is often needed to obtain significant protective effects [44][45][46][47].…”

Immunosuppressive effects of an ethyl acetate extract from Urtica dentata Hand on skin allograft rejection

“…FACS was performed as described by Krueger et al (2003). Cells (2 × 10 6 cells/ml) were sustained at 4 • C for 30 min with the following antibodies diluted in PBS containing 5% FCS: PEconjugated anti-CD86, PE-conjugated anti-MHC-II.…”

Histopathological Changes in Insulin, Glucagon and Somatostatin Cells in the Islets of NOD Mice During Cyclophosphamide-accelerated Diabetes: A Combined Immunohistochemical and Histochemical Study

“…Cyclophosphamide was first employed by Harada and Makino to synchronise and accelerate diabetes in the NOD mouse and was subsequently proposed to have an immunosuppressive effect also (Harada & Makino 1984, Yasunami & Bach 1988. Numerous investigators have since employed this model to study the immunopathogenesis of IDDM and to evaluate a range of preventive therapies (Bacelj et al 1989, Kay et al 1991, Zhang et al 1993, Rothe et al 1994, Cailleau et al 1997, Reddy et al 1999, 2001a, b, Maron et al 2001, Krueger et al 2003. In the cyclophosphamide model, a single injection of the drug to female NOD mice during established insulitis, results in clinical diabetes approximately 14 days later in most animals (Reddy et al 1999).…”