A microsphere-based vaccine prevents and reverses new-onset autoimmune diabetes.
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.
Phillips
Brett
B
Diabetes Institute, Division of Immunogenetics, Department of Pediatrics, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, USA.
Nylander
Karen
K
Harnaha
Jo
J
Machen
Jennifer
J
Lakomy
Robert
R
Styche
Alexis
A
Gillis
Kimberly
K
Brown
Larry
L
Lafreniere
Debra
D
Gallo
Michael
M
Knox
Janet
J
Hogeland
Kenneth
K
Trucco
Massimo
M
Giannoukakis
Nick
N
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
D000906:Antibodies; D018122:B7-1 Antigen; D051940:B7-2 Antigen; D001786:Blood Glucose; D019013:CD40 Antigens; D007328:Insulin; D016376:Oligonucleotides, Antisense