title: A microsphere-based vaccine prevents and reverses new-onset autoimmune diabetes.
abstract: OBJECTIVE
  This study was aimed at ascertaining the efficacy of antisense oligonucleotide-formulated microspheres to prevent type 1 diabetes and to reverse new-onset disease.
  
  
  METHODS
  Microspheres carrying antisense oligonucleotides to CD40, CD80, and CD86 were delivered into NOD mice. Glycemia was monitored to determine disease prevention and reversal. In recipients that remained and/or became diabetes free, spleen and lymph node T-cells were enriched to determine the prevalence of Foxp3(+) putative regulatory T-cells (Treg cells). Splenocytes from diabetes-free microsphere-treated recipients were adoptively cotransferred with splenocytes from diabetic NOD mice into NOD-scid recipients. Live-animal in vivo imaging measured the microsphere accumulation pattern. To rule out nonspecific systemic immunosuppression, splenocytes from successfully treated recipients were pulsed with beta-cell antigen or ovalbumin or cocultured with allogeneic splenocytes.
  
  
  RESULTS
  The microspheres prevented type 1 diabetes and, most importantly, exhibited a capacity to reverse clinical hyperglycemia, suggesting reversal of new-onset disease. The microspheres augmented Foxp3(+) Treg cells and induced hyporesponsiveness to NOD-derived pancreatic beta-cell antigen, without compromising global immune responses to alloantigens and nominal antigens. T-cells from successfully treated mice suppressed adoptive transfer of disease by diabetogenic splenocytes into secondary immunodeficient recipients. Finally, microspheres accumulated within the pancreas and the spleen after either intraperitoneal or subcutaneous injection. Dendritic cells from spleen of the microsphere-treated mice exhibit decreased cell surface CD40, CD80, and CD86.
  
  
  CONCLUSIONS
  This novel microsphere formulation represents the first diabetes-suppressive and reversing nucleic acid vaccine that confers an immunoregulatory phenotype to endogenous dendritic cells.
authors.affiliation: Diabetes Institute, Division of Immunogenetics, Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA.
authors.forename: Brett
authors.initials: B
authors.lastname: Phillips
authors.forename: Karen
authors.initials: K
authors.lastname: Nylander
authors.forename: Jo
authors.initials: J
authors.lastname: Harnaha
authors.forename: Jennifer
authors.initials: J
authors.lastname: Machen
authors.forename: Robert
authors.initials: R
authors.lastname: Lakomy
authors.forename: Alexis
authors.initials: A
authors.lastname: Styche
authors.forename: Kimberly
authors.initials: K
authors.lastname: Gillis
authors.forename: Larry
authors.initials: L
authors.lastname: Brown
authors.forename: Debra
authors.initials: D
authors.lastname: Lafreniere
authors.forename: Michael
authors.initials: M
authors.lastname: Gallo
authors.forename: Janet
authors.initials: J
authors.lastname: Knox
authors.forename: Kenneth
authors.initials: K
authors.lastname: Hogeland
authors.forename: Massimo
authors.initials: M
authors.lastname: Trucco
authors.forename: Nick
authors.initials: N
authors.lastname: Giannoukakis
mesh_terms: D000818:Animals; D000906:Antibodies / Q000008:administration & dosage; D018122:B7-1 Antigen / Q000276:immunology; D051940:B7-2 Antigen / Q000276:immunology; D001786:Blood Glucose / Q000378:metabolism; D019013:CD40 Antigens / Q000276:immunology; D003922:Diabetes Mellitus, Type 1 / Q000097:blood / Q000276:immunology*; D005260:Female; D007328:Insulin / Q000627:therapeutic use; D008213:Lymphocyte Activation / Q000187:drug effects; D051379:Mice; D008810:Mice, Inbred C57BL; D016688:Mice, Inbred NOD; D008863:Microspheres; D016376:Oligonucleotides, Antisense / Q000627:therapeutic use; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology
chemical_list: D000906:Antibodies; D018122:B7-1 Antigen; D051940:B7-2 Antigen; D001786:Blood Glucose; D019013:CD40 Antigens; D007328:Insulin; D016376:Oligonucleotides, Antisense