Interleukin-10 plasmid construction and delivery for the prevention of type 1 diabetes.
Studies of animals with spontaneous autoimmune diabetes have revealed that autoreactive T cells that mediate islet beta cell destruction can be manipulated by the administration of Th(2) cytokines. In this article, the effect of interleukin-10 (IL-10) gene delivery was evaluated in vitro and in vivo with a novel IL-10 plasmid, pSI-IL-10-NFkappaB. In pSI-IL-10-NFkappaB, the expression of the IL-10 gene was driven by the SV40 promotor/enhancer. The nuclear factor kappaB (NFkappaB) binding sites were also introduced to facilitate nuclear transport of the plasmid in the cell. In vitro transfection assay with pSI-IL-10-NFkappaB showed a similar expression level of IL-10 to the plasmid without NFkappaB binding sites (pSI-IL-10). pSI-IL-10-NFkappaB and pSI-IL-10 were intravenously injected into 5-week-old nonobese diabetic (NOD) mice using polyethylenimine (PEI) as a gene carrier. Both groups had persistent gene expression, longer than 5 weeks, and secreted the similarly high IL-10 serum levels. Interestingly, the degree of insulitis in the pSI-IL-10-NFkappaB group was improved over the pSI-IL-10 group, PEI-only group, and noninjected controls. The serum glucose levels showed that single injection of pSI-IL-10-NFkappaB prevented the development of diabetes in 100% of the pSI-IL-10-NFkappaB-injected animals (5/5), while that of pSI-IL-10 prevented diabetes in 40% of the treated animals (2/5). These results suggest that pSI-IL-10-NFkappaB with PEI can effectively reduce the incidence of insulitis and type 1 diabetes in NOD mice.
Lee
Minhyung
M
Department of Bioengineering, College of Engineering, Hanyang University Hospital, 249-1 Kyomun-dong, Kuri, Kyunggi-do 471-020, Korea.
Park
Hyewon
H
Youn
Jeehee
J
Oh
Eun Taex
ET
Ko
Kyungsoo
K
Kim
Sungwan
S
Park
Yongsoo
Y
D000818:Animals; D001665:Binding Sites; D001786:Blood Glucose / Q000032:analysis; D002460:Cell Line; D003922:Diabetes Mellitus, Type 1 / Q000517:prevention & control*; D005260:Female; D018014:Gene Transfer Techniques; D006801:Humans; D066298:In Vitro Techniques; D016753:Interleukin-10 / Q000235:genetics* / Q000378:metabolism / Q000627:therapeutic use*; D051379:Mice; D016688:Mice, Inbred NOD; D010957:Plasmids / Q000235:genetics*; D011094:Polyethyleneimine / Q000378:metabolism; D011485:Protein Binding; D014162:Transfection
D001786:Blood Glucose; D016753:Interleukin-10; D011094:Polyethyleneimine