Research Article
Morozova OV
Pritchina TN
Isaeva EI
Recieved on: 2025-03-27, Accepted on: 2025-05-23, Published on: 2025-06-02
Intracellular delivery of natural and artificial nanomaterials is based on endocytosis, phagocytosis, macropinocytosis and pinocytosis. The endocytosis is the main way for nanomaterials of 50-500 nm even in the absence of receptors. In endocytic course both natural, environmental and artificial nanomaterials become trapped in endosomes and later in lysosomes, where lysosomal enzymes hydrolyze foreign proteins, nucleic acids, carbohydrates and lipids. Antigen presentation of the short hydrolyzed fragments with the major histocompatibility complex (MHC) induces innate immunity with cytokine gene expression.
Objective: analysis of human interferon gene expression in cells and culture media in the presence of protein nanoparticles, extracellular vesicles and post infection with Mycoplasma spp., influenza A virus (IAV) and respiratory syncytial virus (RSV).
Methods: Protein nanoparticles (NP) were fabricated by nanoprecipitation of bovine serum albumin (BSA) and human immunoglobulins (Ig) from their solutions in fluoroalcohol. Extracellular vesicles (EV) were isolated from the conditioned culture media of human embryonic and cancer cells by means of differential centrifugation at 21,000 g. Interferon (IFN) α, β, γ and λ RNA were detected using reverse transcription with subsequent quantitative real-time PCR (RT2-PCR) with fluorescent hydrolysis probes. Interferons were also revealed by using ELISA.
Results: NP consisting of non-immunogenic proteins (albumin and human IgG) induced human IFN α, β and λ, but not IFN γ gene expression. Neither IFN RNA of type I and II nor corresponding proteins were detected in the presence of EV due to flexible conformations of lipids and trace amounts of integral transmembrane proteins in cellular membranes. The only exception was IFN λ detected by ELISA with concentrations 60-70 pg/ml in culture media of the human embryonal cells after addition of heterologous EV isolated from human cancer cells. Both viral and mycoplasma infections resulted in IFN gene expression that inhibited subsequent additional viral infections.
Conclusion: Th1 polarized innate immunity with IFN production resulted in protection against further viral infection.