Adenoviral vector-mediated interleukin-10 expression in vivo: intramuscular gene transfer inhibits cytokine responses in endotoxemia.
Interleukin-10 (IL-10) is a potent anti-inflammatory/immune cytokine and has received growing attention for its therapeutic potential. To aid therapeutic studies of IL-10 in vivo, a replication-deficient adenoviral vector expressing mouse IL-10 was constructed and characterized. The transgene protein IL-10 was shown to markedly inhibit endotoxin-induced tumor necrosis factor alpha (TNF alpha) production by mouse and rat macrophages in vitro. Intramuscular injection of this vector in mice resulted in efficient expression of transgene mRNA in the muscle and active release of IL-10 protein into the bloodstream. To investigate the therapeutic potential of IL-10 using this vector, endotoxemia was induced by intraperitoneal injection of a sublethal dose of endotoxin. Expression of TNF alpha and IL-6 mRNA in the lung, spleen and heart and the circulating levels of these cytokines markedly increased in endotoxemia. This endotoxin-induced TNF alpha and IL-6 up-regulation was however suppressed in mice expressing IL-10 after intramuscular gene transfer. While cytokine gene expression was inhibited to varying degrees in different organs, a maximal reduction was seen in the lung, thus also indicating the efficacy of systemic IL-10 gene product at multiple tissue sites. Finally, we provided evidence that only when present in abnormally high concentrations in the circulation following intraperitoneal gene delivery, IL-10 by itself had some toxic effects of transient nature, primarily manifested by acute phase reaction and hemostatic disturbance. Thus, our studies demonstrate the usefulness of adenoviral vectors for therapeutic applications of IL-10 in vivo.
Xing
Z
Z
Department of Pathology, McMaster University, Hamilton, Ontario, Canada.
Ohkawara
Y
Y
Jordana
M
M
Graham
F L
FL
Gauldie
J
J
D000210:Acute-Phase Reaction / Q000209:etiology; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D019446:Endotoxemia / Q000276:immunology / Q000628:therapy*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000627:therapeutic use*; D006470:Hemorrhage / Q000209:etiology / Q000473:pathology; D006801:Humans; D007273:Injections, Intramuscular; D016753:Interleukin-10 / Q000009:adverse effects / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats
D016207:Cytokines; D012333:RNA, Messenger; D016753:Interleukin-10