Interleukin-4 or interleukin-10 expressed from adenovirus-transduced syngeneic islet grafts fails to prevent beta cell destruction in diabetic NOD mice.
BACKGROUND
We performed ex vivo adenoviral gene transfer in a mouse pancreatic islet transplant model to test the efficacy of this expression system. We then determined whether adenoviral-mediated expression of mouse interleukin (IL) 4 or IL-10 from transduced syngeneic islet grafts could prevent disease recurrence in diabetic nonobese diabetic (NOD) mice.


METHODS
An adenoviral vector expressing beta-galactosidase (AdCMV betaGal) was used to transduce BALB/c islets (2.5 x 10(3) plaque-forming units/islet), which were analyzed for glucose responsiveness, islet cell recovery, and efficiency of gene transfer. In vivo function and reporter gene expression were examined with AdCMV betaGal-transduced islet grafts in alloxan-induced diabetic syngeneic recipients. Adenoviruses expressing either IL-4 or IL-10 were used in a similar fashion to infect NOD islets, which were characterized in vitro, as well as transplanted into diabetic syngeneic recipients.


RESULTS
In vitro functional studies showed no significant difference between control or transduced islets, with 50+/-4% of AdCMV betaGal-infected islet cells staining positive for beta-galactosidase. Transplant recipients became nomoglycemic within 48 hr after transplant, and, although beta-galactosidase expression decreased over time, it was detectable in the graft for up to 8 weeks. Despite the nanogram quantities of IL-4 or IL-10 produced/day from each graft equivalent in vitro, transduced and transplanted NOD islets failed to prevent disease recurrence.


CONCLUSIONS
These results suggest that adenoviruses are efficient for at least medium term gene expression from islets in vivo, but neither IL-4 nor IL-10 alone can prevent autoimmune disease recurrence in NOD mice.
Smith
D K
DK
Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Canada.
Korbutt
G S
GS
Suarez-Pinzon
W L
WL
Kao
D
D
Rajotte
R V
RV
Elliott
J F
JF
D000256:Adenoviridae; D000818:Animals; D003587:Cytomegalovirus; D003922:Diabetes Mellitus, Type 1 / Q000601:surgery*; D005822:Genetic Vectors; D005947:Glucose / Q000494:pharmacology; D006085:Graft Survival / Q000276:immunology / Q000502:physiology*; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D016753:Interleukin-10 / Q000096:biosynthesis*; D015847:Interleukin-4 / Q000096:biosynthesis*; D007515:Islets of Langerhans / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D016381:Islets of Langerhans Transplantation / Q000276:immunology* / Q000473:pathology / Q000502:physiology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D016688:Mice, Inbred NOD; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D014162:Transfection; D014185:Transplantation, Isogeneic; D001616:beta-Galactosidase / Q000096:biosynthesis
D007328:Insulin; D011993:Recombinant Fusion Proteins; D016753:Interleukin-10; D015847:Interleukin-4; D001616:beta-Galactosidase; D005947:Glucose