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Protection from experimental endotoxemia by a recombinant adeno‐associated virus encoding interleukin 10
Author(s) -
Yamano Seiichi,
Scott Dorothy E.,
Huang LiYun,
Mikolajczyk Malgorzata,
Pillemer Stanley R.,
Chiorini John A.,
Golding Basil,
Baum Bruce J.
Publication year - 2001
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.213
Subject(s) - lipopolysaccharide , in vivo , spleen , adeno associated virus , recombinant dna , cytokine , immunology , genetic enhancement , vector (molecular biology) , septic shock , interleukin 10 , medicine , biology , pharmacology , virus , in vitro , gene , sepsis , biochemistry , microbiology and biotechnology
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 (10 10 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. Copyright © 2001 John Wiley & Sons, Ltd.