Ex vivo gene transfer of viral interleukin-10 to BB rat islets: no protection after transplantation to diabetic BB rats.
Allogeneic and autoimmune islet destruction limits the success of islet transplantation in autoimmune diabetic patients. This study was designed to investigate whether ex vivo gene transfer of viral interleukin-10 (vIL-10) protects BioBreeding (BB) rat islets from autoimmune destruction after transplantation into diabetic BB recipients. Islets were transduced with adenoviral constructs (Ad) expressing the enhanced green fluorescent protein (eGFP), alpha-1 antitrypsin (AAT) or vIL-10. Transduction efficiency was demonstrated by eGFP-positive cells and vIL-10 production. Islet function was determined in vitro by measuring insulin content and insulin secretion and in vivo by grafting AdvIL-10-transduced islets into syngeneic streptozotocin (SZ)-diabetic, congenic Lewis (LEW.1 W) rats. Finally, gene-modified BB rat islets were grafted into autoimmune diabetic BB rats. Ad-transduction efficiency of islets increased with virus titre and did not interfere with insulin content and insulin secretion. Ad-transduction did not induce Fas on islet cells. AdvIL-10-transduced LEW.1 W rat islets survived permanently in SZ-diabetic LEW.1 W rats. In diabetic BB rats AdvIL-10-transduced BB rat islets were rapidly destroyed. Prolongation of islet culture prior to transplantation improved the survival of gene-modified islets in BB rats. Several genes including those coding for chemokines and other peptides associated with inflammation were down-regulated in islets after prolonged culture, possibly contributing to improved islet graft function in vivo. Islets transduced ex vivo with vIL-10 are principally able to cure SZ-diabetic rats. Autoimmune islet destruction in diabetic BB rats is not prevented by ex vivo vIL-10 gene transfer to grafted islets. Graft survival in autoimmune diabetic rats may be enhanced by improvements in culture conditions prior to transplantation.
Kuttler
Beate
B
Institute of Physiology, Medical School, Ernst-Moritz-Arndt-University of Greifswald, Greifswalderstrasse 11c, D-17495 Karlsburg, Germany. kuttler@uni-greifswald.de
Wanka
Heike
H
Klöting
Nora
N
Gerstmayer
Bernhard
B
Volk
Hans-Dieter
HD
Sawitzki
Birgit
B
Ritter
Thomas
T
D000256:Adenoviridae; D000818:Animals; D015551:Autoimmunity / Q000276:immunology; D001786:Blood Glucose / Q000032:analysis; D016000:Cluster Analysis; D003921:Diabetes Mellitus, Experimental / Q000473:pathology*; D005434:Flow Cytometry; D005822:Genetic Vectors; D004854:Herpesvirus 4, Human / Q000378:metabolism*; D007328:Insulin / Q000378:metabolism; D016753:Interleukin-10 / Q000235:genetics*; D007515:Islets of Langerhans / Q000378:metabolism* / Q000473:pathology*; D016381:Islets of Langerhans Transplantation; D020411:Oligonucleotide Array Sequence Analysis; D051381:Rats; D011913:Rats, Inbred BB; D011917:Rats, Inbred Lew; D013311:Streptozocin; D014161:Transduction, Genetic; D019014:fas Receptor / Q000378:metabolism
D001786:Blood Glucose; D007328:Insulin; D019014:fas Receptor; D016753:Interleukin-10; D013311:Streptozocin