Adeno-associated virus vector-mediated IL-10 gene delivery prevents type 1 diabetes in NOD mice

Abstract

The development of spontaneous autoimmune diabetes in nonobese diabetic (NOD) mice provides for their use as a model of human type 1 diabetes. To test the feasibility of muscle-directed gene therapy to prevent type 1 diabetes, we developed recombinant adeno-associated virus (rAAV) vectors containing murine cDNAs for immunomodulatory cytokines IL-4 or IL-10. Skeletal muscle transduction of female NOD mice with IL-10, but not IL-4, completely abrogated diabetes. rAAV-IL-10 transduction attenuated the production of insulin autoantibodies, quantitatively reduced pancreatic insulitis, maintained islet insulin content, and altered splenocyte cytokine responses to mitogenic stimulation. The beneficial effects were host specific, as adoptive transfer of splenocytes from rAAV IL-10-treated animals rapidly imparted diabetes in naive hosts, and the cells contained no protective immunomodulatory capacity, as defined through adoptive cotransfer analyses. These results indicate the utility for rAAV, a vector with advantages for therapeutic gene delivery, to transfer immunoregulatory cytokines capable of preventing type 1 diabetes. In addition, these studies provide foundational support for the concept of using immunoregulatory agents delivered by rAAV to modulate a variety of disorders associated with deleterious immune responses, including allergic reactions, transplantation rejection, immunodeficiencies, and autoimmune disorders.

IL-10 delivery by AAV5 vector attenuates inflammation in mice with pseudomonas pneumonia

“…59 To facilitate pseudotyping, pAAV.Cb-mIL10 was co-transfected with pXYZ5, an adenoviral helper plasmid with AAV2 rep/AAV5 cap. 60 Anion-exchange chromatography was used to purify the AAV5 pseudotyped vector, and the dot-blot assay was used to determine the titer of rAAV virions harboring vector genomes.…”

Immunity to adeno-associated virus serotype 2 delivered transgenes imparted by genetic predisposition to autoimmunity

“…We recently observed muscle inflammation in nonobese diabetic (NOD) mice, a model for autoimmune type I diabetes, that had received interleukin-4 (IL-4) and interleukin-10 (IL-10) therapy via intramuscular AAV2-IL-4 or AAV2-IL-10 injections. 23 This further raised the question as to how genetic predisposition to autoimmunity affects the effectiveness of AAV gene therapy.…”

Adeno-associated virus-mediated human IL-10 gene transfer suppresses the development of experimental autoimmune orchitis

“…13,24 The effects of IL-10 on autoimmune diseases using experimental models have been studied for years, but the results are somewhat confusing. In nonobese diabetic mice, IL-10 supplementation prevented autoimmune diabetes in some experimental protocols, 11,12 whereas augmented it in others. 25 Similarly, the results of a recent study by Kaneko et al 26 of IL-10 in EAO differ from our present results.…”

“…The fact that IL-10 is an immunoregulatory cytokine 10 with potential therapeutic value for organ-specific autoimmune inflammatory disease [11][12][13][14] strongly suggest that IL-10 has the potential to suppress autoimmune inflammation occurring in the testes.…”