In vitro cell-based Hyperurecemia-hemotest bioassay for evaluation of cytokine status of patients with gouty arthritis

Abstract

Aim. To develop an in vitro method for assessing the activity of the inflammasome under conditions of hyperuricemic stimulation of inflammatory interleukins. Material and methods. Whole blood cells of donors diluted with RPMI were cultured in vitro in the presence of different concentrations of uric acid. The production of cytokines in the cell growth media of hematopoietic cells stimulated with uric acid was evaluated using an enzyme-linked immunosorbent assay (ELISA). Results. Simulating the hyperuricemia in vivo, an in vitro cell-based bioassay was developed to stimulate blood cells of individual donors with uric acid. Using the developed in vitro Hyperuricemia-hemotest bioassay, quantitative differences were found in the production of inflammatory cytokines by the blood cells of potentially healthy donors and patients with hyperuricemia and gouty arthritis. Conclusion. As a new approach in personalized diagnostics, a hyperuricemic (HU)-hemotest system was developed, which can serve as an in vitro cell model for studying the activation of inflammasome by inflammatory signaling molecules. Using this test-system, it was shown that in vitro blood cells of patients sensitized under in vivo conditions with factors specific for hyperuricemia and gouty arthritis react differently to the presence of uric acid than cells of potentially healthy donors.