Studying expression of IL-1β gene under the action of siRNA complexes with anti-influenza effect

Abstract

Influenza is one of the most urgent global health problems today. The influenza virus has immunosuppressive properties, which can lead to the development of secondary immunodeficiencies, interfering with the functioning of the interferon system activation, thus leading to impaired production of pro-inflammatory cytokines. IL-1 is the most important player in development of antiviral immunity. This cytokine plays an important role in boosting the expression of the MCP-1 and MCP-3 genes and maturation of macrophages and dendritic cells. Induction of IL-1 production occurs due to interaction of the ligand with Toll-like receptors. Currently, there is a lot of drugs aimed at the prevention and treatment of influenza infection. However, their use in some cases is difficult due to high mutational variability of the influenza virus, thus making it resistant to these drugs. Therefore, the issue of developing and creating effective methods to combat such infections is of particular importance. A promising approach to the treatment and prevention of viral respiratory infections may be connected with RNA interference. This process consists of degradation of foreign mRNA by small interfering RNA (siRNA) molecules. The aim of the present study was to evaluate expression of the IL-1 gene upon transfection of miRNA complexes directed to the cellular FLT4, Nup98, Nup205 genes. Evaluation of changed viral reproduction was carried out using titration by CPE virus-containing fluid. Expression level of the IL-1 gene was determined by means of real-time RT-PCR. Assessment of the changes in viral reproduction allowed us to reveal that the use of all the miRNA complexes directed to the cellular genes lead to a significant decrease in viral reproduction on the 1st day after infection. Usage of Nup205 + FLT4 and FLT4 + Nup205 + Nup98 complexes proved to cause a decrease in viral reproduction on the second day as well (p 0.05), as compared with nonspecific and viral controls. When analyzing expression profile of the IL-1 gene, an increase in its expression was observed on the 1st day for all miRNA complexes and on the 2nd and 3rd days for the Nup98 + FLT4 and Nup205 + Nup98 complexes. In the course of the study, it was found that suppression of the cellular genes FLT4, Nup98 and Nup205 activities, which are necessary for viral reproduction, led to a significant decrease in viral activity and an increase in IL-1 expression.