Cell type specific and glucose responsive expression of interleukin-4 by using insulin promoter and water soluble lipopolymer.
For gene therapy, tissue targeting of gene delivery systems is required for the maximum efficiency. In this study, we constructed pRIP-IL4 in which the expression of interleukin-4 (IL-4) was driven by the rat insulin promoter. WSLP-pRIP-IL4 complex was characterized by pancreas beta-cell specific and glucose responsive expression of IL-4. pRIP-IL4 was completely retarded at a 6:1 or higher N/P (nitrogen atom of WSLP/phosphate of plasmid) ratio in 1% agarose gel. In addition, WSLP protected plasmid DNA from DNase I for more than 1 h. In cytotoxicity assay, WSLP showed less cytotoxicity than PEI (25000 Da) to mouse insulinoma (MIN6) cells. ELISA showed that pRIP-IL4 expressed much higher levels of IL-4 in MIN6 cells than in NIH3T3 cells. The expression level of IL-4 by pRIP-IL4 increased with increasing concentration of glucose. Also, IL-4 was expressed in a dose-dependent manner. This WSLP-pRIP-IL4 system will be useful in the development of a pancreas specific expression system for the prevention of diabetes without systemic side effects.
Lee
M
M
Center for Controlled Chemical Delivery, University of Utah, 30 S 2000 E, RM 201, Salt Lake City, UT 84112-5820, USA. minhyung.lee@deans.pharm.utah.edu
Han
S
S
Ko
K S
KS
Kim
S W
SW
D000818:Animals; D002460:Cell Line; D015316:Genetic Therapy; D005947:Glucose / Q000494:pharmacology*; D007328:Insulin / Q000235:genetics*; D015847:Interleukin-4 / Q000008:administration & dosage* / Q000096:biosynthesis; D051379:Mice; D011094:Polyethyleneimine / Q000008:administration & dosage; D011108:Polymers / Q000008:administration & dosage; D011401:Promoter Regions, Genetic; D014162:Transfection
D007328:Insulin; D011108:Polymers; D015847:Interleukin-4; D011094:Polyethyleneimine; D005947:Glucose