Prolonged xenograft survival of islets infected with small doses of adenovirus expressing CTLA4Ig.
BACKGROUND
Systemic administration of the inhibitor of costimulation, CTLA4Ig, has been shown to prolong islet graft survival. The purpose of this study was to compare local and systemic expression of murine CTLA4Ig in transplants of rat islets into mice.


METHODS
Murine CTLA4Ig was made by joining two polymerase chain reaction products, the extracellular portion of CTLA4 and the Fc portion of IgG2a. Recombinant adenovirus expressing CTLA4Ig (AdCTLA4Ig) was generated using the strategy of Cre-lox recombination. Isolated rat islets infected with AdCTLA4Ig at multiplicities of infection (MOIs) ranging from 0.1 to 10 were transplanted into streptozocin diabetic male B6AF1 mice. Control islets were mock infected or infected with AdLacZ or AdsIg, a recombinant adenovirus expressing only the Fc portion of IgG2a. Also, AdCTLA4Ig and control viruses were injected intramuscularly into mouse transplant recipients at the time of islet transplantation to provide CTLA4Ig systemically.


RESULTS
Control islets transplanted into diabetic mice were rejected in 13-17 days. Islets infected with AdCTLA4Ig had dose-dependent prolongation of graft survival. Prolonged survival was even found with very low MOIs of 0.1 and 0.5, with survivals of 24+/-4.2 and 25+/-2.2 days, respectively. Survival with an MOI of 10 was 39+/-8.7 days. With intramuscular injection, no prolongation was found at the lowest relative MOIs of 0.2 and 1, but there was dose-dependent prolongation of graft survival with larger doses. At the highest relative MOI of 400, survival was prolonged to 58+/-10 days.


CONCLUSIONS
Rat islets infected with AdCTLA4Ig transplanted into mice had prolonged graft survival. Prolonged survival with MOIs as low as 0.1 and 0.5 indicates that only a minority of islet cells need to express CTLA4Ig to exert an effect. Moreover, the results suggest that the improved islet graft survival is due to a local influence of CTLA4Ig.
Department of Genetics, Harvard Medical School and Howard Hughes Medical Institute, The Children's Hospital, Boston, Massachusetts 02115, USA. feng@helix.mgh.harvard.edu
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Feng
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Quickel
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Hollister-Lock
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McLeod
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Bonner-Weir
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Mulligan
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Weir
D000069594:Abatacept; D000257:Adenoviridae Infections / Q000235:genetics*; D000818:Animals; D015703:Antigens, CD; D000943:Antigens, Differentiation / Q000235:genetics* / Q000494:pharmacology; D060908:CTLA-4 Antigen; D015870:Gene Expression / Q000187:drug effects; D006085:Graft Survival / Q000187:drug effects; D018796:Immunoconjugates; D007141:Immunoglobulin Fc Fragments / Q000494:pharmacology; D007166:Immunosuppressive Agents / Q000494:pharmacology; D007273:Injections, Intramuscular; D016381:Islets of Langerhans Transplantation / Q000276:immunology*; D007763:Lac Operon / Q000235:genetics; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D051381:Rats; D017207:Rats, Sprague-Dawley; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000494:pharmacology; D014183:Transplantation, Heterologous / Q000276:immunology*; D001616:beta-Galactosidase / Q000235:genetics
D015703:Antigens, CD; D000943:Antigens, Differentiation; D060908:CTLA-4 Antigen; C556707:Ctla4 protein, mouse; C556708:Ctla4 protein, rat; D018796:Immunoconjugates; D007141:Immunoglobulin Fc Fragments; D007166:Immunosuppressive Agents; D011993:Recombinant Fusion Proteins; D000069594:Abatacept; D001616:beta-Galactosidase