Gene transfer with IL-4 and IL-13 improves survival in lethal endotoxemia in the mouse and ameliorates peritoneal macrophages immune competence

Abstract

Systemic anti-cytokine therapies have been unsuccessful in preventing mortality from gram-negative bacteremia in humans partly because of the failure to neutralize pro-inflammatory cytokines at sites of exaggerated production. In an attempt to deliver anti-inflammatory cytokines to organs directly, gene transfer was employed. Thirty-six BALB/c mice were injected intraperitoneally with cationic liposomes containing plasmids encoding the human interleukin-4 (hIL-4) or IL-13 gene. Both, hIL-4 and hIL-13 mRNA were detected by reverse transcription-polymerase chain reaction analysis in the liver and the spleen of the animals. Fourty-eight hours after the in vivo gene transfer, these 36 mice and 18 mock-transfected mice, were challenged with a lethal dose of E. coli lipopolysaccharide with D-galactosamine (D-GalN). Gene transfer with hIL-4 reduced the serum tumor necrosis factor (TNF)-alpha production in response to endotoxin/D-GalN by 80% from 113.1 pg/ml in mock-transfected animals to 22.2 pg/ml (p < 0.05); human IL-13 gene transfer reduced serum TNF-alpha levels by 90% (113.1 pg/ml to 11.6 pg/ml; p < 0.05). Survival was improved from 20% to over 83% in both treatment groups (p < 0.001). Our data demonstrate a potent in vivo anti-inflammatory action of both IL-4 and IL-13. In addition, the immune functions of peritoneal macrophages are significantly ameliorated in both treatment groups, with IL-13 demonstrating better macrophage immune modulation than IL-4 (p < 0.05).

The Involvement of Tyrosine Kinases, Cyclic AMP/Protein Kinase A, and p38 Mitogen-Activated Protein Kinase in IL-13-Mediated Arginase I Induction in Macrophages: Its Implications in IL-13-Inhibited Nitric Oxide Production

“…Therefore, we conclude that the IL-13-reduced NO production is regulated by at least two mechanisms, the up-regulated arginase activity and the down-regulated iNOS expression. These results also suggest that the induced arginase, by inhibiting NO production from activated macrophages, may contribute at least in part to the observed anti-inflammatory effect of IL-13 in vivo (42). It is worth noting that L-norvaline specifically inhibits arginase activity at the enzyme kinetic level (15,26), and it does not affect iNOS expression, as shown in the present study.…”

IL-9 Protects Mice from Gram-Negative Bacterial Shock: Suppression of TNF-α, IL-12, and IFN-γ, and Induction of IL-10

“…This concept is supported by much evidence derived from studies in animal and in human systems in which the systemic injection of LPS or live bacteria or the presence of bacterial infection is associated with the sustained production of proinflammatory cytokines. Various studies in animal models have demonstrated the protective effect of anti-cytokine maneuvers, including the administration or induced expression of the Th2 cytokines IL-10 (17-19) and IL-4 (20). In addition, the antiinflammatory cytokine IL-13 has been found to provide protection from LPS-induced endotoxemia in a manner that is similar to but distinct from that of IL-10 (23).…”

“…IL-10 can act as a potent modulator of cytokine production and lethality in LPS-induced pathology (17)(18)(19). Gene transfer with IL-4 improves survival in lethal endotoxemia (20), and selective, compartmentalized blockade of TNF-␣ overproduction by IL-4 enhances pulmonary clearance of Pseudomonas aeruginosa in mice (21). We have recently shown that rIL-4 may protect animals from death in a toxic shock model with the bacterium, correlating with decreased levels of circulating TNF-␣ soon after challenge (22).…”

“…We have recently shown that rIL-4 may protect animals from death in a toxic shock model with the bacterium, correlating with decreased levels of circulating TNF-␣ soon after challenge (22). IL-13, a cytokine that shares anti-inflammatory properties with IL-4 and IL-10, protects mice from LPS-induced lethal endotoxemia (20,23).…”

Liposome-mediated, nonviral gene transfer induces a systemic inflammatory response which can exacerbate pre-existing inflammation

“…13,14 Several groups have demonstrated that liposome and plasmid-mediated expression of either IL-10, soluble TNF receptors, COX-2, IL-4 or IL-11 could reduce the inflammatory consequences of endotoxin shock or caerulein-induced pancreatitis. [15][16][17][18] However, these therapies were generally administered hours or days before the induction of experimental inflammation, and the resulting inflammatory response to the administered liposomes and plasmid DNA may have induced tolerance in these animals and contributed to the improved morbidity and mortality. The present study examined whether the inflammatory response to administered liposomes and plasmid DNA could affect the outcome during experimental inflammation.…”