Lentiviral-vector-mediated expression of murine IL-1 receptor antagonist or IL-10 reduces the severity of endotoxin-induced uveitis

Abstract

Uveitis is a sight threatening inflammatory disorder that remains a significant cause of visual loss. We investigated lentiviral gene delivery of IL-1ra or IL-10 to ameliorate murine endotoxin-induced uveitis (EIU). An HIV-1-based vector containing the mIL-1ra or mIL-10 cDNA demonstrated high expression of biologically active cytokine. Following administration of Lenti.GFP into the anterior chamber, transgene expression was observed in corneal endothelial cells, trabecular meshwork and iris cells. To treat EIU, mice were injected with Lenti.IL-1ra, Lenti.IL-10 or a combination of these. EIU was induced 14 days after vector administration and mice were culled 12 h following disease induction. Lenti.IL-1ra or Lenti.IL10 treated eyes showed significantly lower mean inflammatory cell counts in the anterior and posterior chambers compared with controls. The aqueous total protein content was also significantly lower in treated eyes, demonstrating better preservation of the blood-ocular barrier. Furthermore, the treated eyes showed less in vivo fluorescein leakage from inner retinal vessels compared with controls. The combination of both IL-1ra and IL-10 had no additive effect. Thus, lentiviral gene delivery of IL-1ra or IL-10 significantly reduces the severity of experimental uveitis, suggesting that lentiviral mediated expression of immunomodulatory genes in the anterior chamber offers an opportunity to treat uveitis.

Impact of IL-1 signalling on experimental uveitis and arthritis

“…Given the presence of the receptor and the increased agonist concentrations, the finding that IL-1R deficiency did not significantly affect the severity of uveitis was somewhat surprising, but not completely unexpected. This observation is consistent with previous reports of EIU in rabbits,39 but is not in complete agreement with a report demonstrating that IL-1Ra overexpression in mouse eyes infected with a lentiviral expression vector suppresses EIU 40. Methods for the latter report differed substantially from ours; those authors used four times the amount of LPS and performed multiple LPS injections, suggesting their observation may be dependent on the high doses of LPS.…”

Effects of kefir supernatant and lactic acid bacteria isolated from kefir grain on cytokine production by macrophage

“…3). IL-10 could contribute an inhibitory effect to IL-1b (Trittibach et al, 2008). Thus, both cytokines could not be detected after 24 h of co-culture.…”

Intraocular Delivery of miR-146 Inhibits Diabetes-Induced Retinal Functional Defects in Diabetic Rat Model

“…Our data suggests that lenti-miR-146a delivered functional miR-146a, which sustained its function for at least 3 months in vivo. Using similar lentiviral constructs, robust transgene expression in vivo has been reported in RPE, [79][80][81] photoreceptors, 79,80,82 cornea endothelial cells, 81,83 neurons in the brain, 84,85 and so on, as early as 4 days after viral injection, 83 lasting as long as 3 months 82,84 ; and the beneficial effect of the transgene can persist as long as 7 months. 80 Whether long-term delivery of miR-146 can be achieved by a single injection of lenti-miR-146a still needs to be determined in future studies.…”