Prevention of autoimmune insulitis by delivery of a chimeric plasmid encoding interleukin-4 and interleukin-10.
The combined administration of interleukin-4 (IL-4) and interleukin-10 (IL-10) expression plasmids has demonstrated synergistic effects on the prevention of autoimmune diabetes. To this end, we constructed a co-expression 'chimeric' plasmid, pCMV-IL4-IL10, in which the expression of IL-4 and IL-10 was driven by two separate CMV immediate early promoters by using the biodegradable polymer, poly[alpha-(4-aminobutyl)-L-glycolic acid] (PAGA) as a gene carrier to optimize gene delivery. In vitro transfection assays of the chimeric plasmid in 293T cells showed higher expression levels as well as dose dependence than the single gene expression plasmids. To evaluate the in vivo efficacy of the chimeric plasmid, the pCMV-IL4-IL10/PAGA complex was intravenously injected into 4-week-old non-obese diabetic (NOD) mice and compared to the co-administration group. While both groups had persistent gene expression longer than 5 weeks, the IL-4 and IL-10 serum levels of the chimeric group were higher than those in the co-administration group. Furthermore, the degree of insulitis in the chimeric group was improved over both the co-administration and non-injected control groups. These results suggest that the chimeric IL-4 and IL-10 expression plasmid can effectively reduce the incidence of autoimmune insulitis.
Lee
Minhyung
M
Center for Controlled Chemical Delivery, 30 S 2000 East RM 201, Department of Pharmaceutics and Pharmaceutical Chemistry, University of Utah, Salt Lake City 84112-5820, USA.
Ko
Kyung Soo
KS
Oh
Seungjoon
S
Kim
Sung Wan
SW
D000818:Animals; D001327:Autoimmune Diseases / Q000517:prevention & control*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015316:Genetic Therapy; D006801:Humans; D007328:Insulin / Q000276:immunology*; D016753:Interleukin-10 / Q000235:genetics*; D015847:Interleukin-4 / Q000235:genetics*; D051379:Mice; D016688:Mice, Inbred NOD; D010957:Plasmids / Q000235:genetics*; D011100:Polyglycolic Acid / Q000031:analogs & derivatives* / Q000138:chemical synthesis; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D014162:Transfection
D007328:Insulin; D011993:Recombinant Fusion Proteins; C414262:poly((4-aminobutyl)glycolic acid); D016753:Interleukin-10; D015847:Interleukin-4; D011100:Polyglycolic Acid