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Development of adenovirus vectors encoding rat complement regulators for use in therapy in rodent models of
Y McGrath1, G W Wilkinson, O B Spiller
1Department of Medical Biochemistry, University of Wales College of Medicine, Cardiff, United Kingdom.
Abstract:
C activation has been implicated in the pathogenesis of numerous inflammatory human diseases and disease models. A therapy based on C inhibition might therefore be of benefit to reduce inflammation and ameliorate disease. C inhibition in vivo can be accomplished by the delivery of soluble recombinant C regulators either systemically or directly to a target site, but effects are transitory. We have developed a strategy for the efficient delivery of the membrane-bound rat C inhibitors, CD59, Crry, and decay-accelerating factor (DAF), using replication-deficient adenovirus vectors with the intention of treating rat models of disease in which C is implicated. The adenovirus recombinants(RAd), RAdCD59, RAdCrry, and RAdDAF, respectively, have been tested for expression and function of the transgene in vitro. Infection of human fetal foreskin fibroblasts resulted in high levels of expression of each of the rat inhibitors. The constructs were also tested for inhibition of rat C-mediated cell lysis and C3b deposition. In a cell lysis assay, each inhibited to varying degrees of efficiency in the order RAdCD59 = RAdDAF > RAdCrry. In a C3b deposition assay, RAdDAF caused a greater reduction in C3b deposition than RAdCrry and RAdCD59 was ineffective. These agents, individually or in combination, provide the tools for testing the effects of prolonged inhibition of C at a target site on the progress of experimental models of disease.
Insights
Adenovirus vectors efficiently deliver rat complement inhibitors CD59, Crry, and decay-accelerating factor (DAF) to target cells. This strategy enables prolonged complement inhibition for treating inflammatory diseases.
Area of Science:
- Immunology
- Molecular Biology
- Gene Therapy
Background:
- Complement (C) activation is central to inflammatory diseases.
- Current C inhibition therapies have transient effects.
- Targeted, prolonged C inhibition is needed for effective treatment.
Purpose of the Study:
- To develop adenovirus vectors for delivering rat C inhibitors (CD59, Crry, DAF).
- To assess the expression and function of these inhibitors in vitro.
- To evaluate their potential for treating C-mediated inflammatory diseases.
Main Methods:
- Constructed replication-deficient adenoviruses (RAd) encoding rat CD59, Crry, and DAF.
- Infected human fibroblasts to confirm transgene expression.
- Assayed inhibition of rat C-mediated cell lysis and C3b deposition.
Main Results:
- High expression levels of CD59, Crry, and DAF achieved.
- RAdCD59 and RAdDAF showed significant inhibition of cell lysis.
- RAdDAF effectively reduced C3b deposition, while RAdCD59 was ineffective.
Conclusions:
- Adenovirus-mediated delivery provides a tool for sustained C inhibition.
- These agents can be used individually or combined for disease modeling.
- This approach holds promise for developing new therapies for inflammatory conditions.