Protection from experimental endotoxemia by a recombinant adeno-associated virus encoding interleukin 10

Seiichi Yamano, Dorothy E. Scott, Li Yun Huang, Malgorzata Mikolajczyk, Stanley R. Pillemer, John A. Chiorini, Basil Golding, Bruce J. Baum

Research output: Contribution to journalArticle

Abstract

Background Interleukin 10 (IL-10) is a homodimeric cytokine that shows considerable clinical promise. Adeno-associated virus (AAV) vectors appear increasingly useful for in vivo gene-transfer applications. Methods A recombinant AAV type 2 vector encoding human IL-10 (rAAVhIL10) was constructed by using an adenoviral-free, three-plasmid co-transfection. Cytokine production was measured by using an enzyme-linked immunosorbent assay. Endotoxic shock was induced by lipopolysaccharide (LPS) injection. Results As media from rAAVhIL10-infected COS cells caused a dose-dependent blockade of IL-12 secretion from spleen cells of IL-10 knockout (KO) mice challenged with Brucella abortus, it was clear that vector-derived hIL-10 was biologically active in vitro. Intravenous or intramuscular administration of relatively modest levels of rAAVhIL10 (1010 genomes) to IL-10 KO mice resulted in hIL-10 secretion into the bloodstream, which, at 8 weeks, gave median serum levels of 0.9 and 0.45 pg/ml, respectively. Acute endotoxic shock led to a 33% mortality rate, and severe morbidity, in control IL-10 KO mice, whereas no mortality and little morbidity were seen in IL-10 KO mice given rAAVhIL10 7 weeks earlier. Conclusions The findings demonstrate that a modest dose of rAAVhIL10 administered in vivo provides long-term protection against LPS-induced endotoxic shock in a murine model. Thus, this vector may be useful for clinical applications requiring sustained IL-10 expression, for example in the treatment of several autoimmune diseases.

Original languageEnglish (US)
Pages (from-to)450-457
Number of pages8
JournalJournal of Gene Medicine
Volume3
Issue number5
DOIs
StatePublished - Sep 2001

Keywords

  • Adeno-associated virus
  • Cytokines
  • Gene transfer
  • Immunoregulation
  • Immunotherapy

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics
  • Drug Discovery
  • Genetics(clinical)

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  • Cite this

    Yamano, S., Scott, D. E., Huang, L. Y., Mikolajczyk, M., Pillemer, S. R., Chiorini, J. A., Golding, B., & Baum, B. J. (2001). Protection from experimental endotoxemia by a recombinant adeno-associated virus encoding interleukin 10. Journal of Gene Medicine, 3(5), 450-457. https://doi.org/10.1002/jgm.213