Cationic Nanomicelles for Delivery of Plasmids Encoding Interleukin-4 and Interleukin-10 for Prevention of Autoimmune Diabetes in Mice

To evaluate the in vivo transfection efficiency of N-acyl derivatives of low-molecular weight chitosan (LMWC) to deliver pVIVO2-mIL4-mIL10 plasmid encoding interleukin-4 (IL-4) and interleukin-10 (IL-10) in multiple, low-dose streptozotocin induced diabetic mouse model. N-acyl LMWC nanomicelles were characterized for size and charge. The pVIVO2-mIL4-mIL10/N-acyl LMWC polyplexes were injected intramuscularly in mice and compared for transfection efficiency with naked DNA and FuGENEA (R) HD. Bicistronic pVIVO2-mIL4-mIL10 plasmid was compared with individual plasmids encoding IL-4 and IL-10 for efficacy. The levels of blood glucose and serum IL-4, IL-10, TNF-alpha and IFN-gamma were monitored. The ability of plasmid administration to protect from insulitis and biocompatibility of N-acyl LMWC were studied. The N-acyl LMWC led to significantly higher (p < 0.05) expression of IL-4 and IL-10 and reduced the levels of blood glucose, TNF-alpha and IFN-gamma, especially in animals treated with pVIVO2-mIL4-mIL10 plasmid. The pancreas of pDNA/N-acyl LMWC polyplex treated animals exhibited protection from insulitis and the delivery systems were found to be biocompatible. N-acyl derivatives of LMWC are efficient and biocompatible gene delivery vectors, and the administration of bicistronic pVIVO2-mIL4-mIL10 plasmid polyplexes can protect the pancreatic islets from insulitis, possibly due to the synergistic effect of IL-4 and IL-10 encoding plasmids.