title: Ex vivo gene transfer of viral interleukin-10 to BB rat islets: no protection after transplantation to diabetic BB rats.
abstract: 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.
authors.affiliation: Institute of Physiology, Medical School, Ernst-Moritz-Arndt-University of Greifswald, Greifswalderstrasse 11c, D-17495 Karlsburg, Germany. kuttler@uni-greifswald.de
authors.forename: Beate
authors.initials: B
authors.lastname: Kuttler
authors.forename: Heike
authors.initials: H
authors.lastname: Wanka
authors.forename: Nora
authors.initials: N
authors.lastname: Klöting
authors.forename: Bernhard
authors.initials: B
authors.lastname: Gerstmayer
authors.forename: Hans-Dieter
authors.initials: HD
authors.lastname: Volk
authors.forename: Birgit
authors.initials: B
authors.lastname: Sawitzki
authors.forename: Thomas
authors.initials: T
authors.lastname: Ritter
mesh_terms: 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
chemical_list: D001786:Blood Glucose; D007328:Insulin; D019014:fas Receptor; D016753:Interleukin-10; D013311:Streptozocin