Activation of Human Dendritic Cells by Nanoparticles Carrying CTL Epitopes of Non-Structural Proteins of Hepatitis C Virus

Abstract

Activation of human dendritic cells by nanoparticles carrying CTL epitopes of non-structural proteins of hepatitis C virus. Archives of Microbiology and Immunology 6 (2022): 39-50. Abstract The aim of this study was to produce immunogenic nanoparticles carrying cytotoxic T lymphocyte (CTL) epitopes of hepatitis C virus non-structural proteins. We have obtained recombinant proteins forming virus-like particles and containing the sequences of self-assembling peptides (SAP), PADRE, and CTL epitopes. Using atomic force microscopy (AFM), the size of thus obtained nanoparticles was shown to be dependent on number of CTL epitopes proliferative activity and interferon-γ (IFN-γ) production by the stimulated T lymphocytes were evaluated by their secondary stimulation with commercially available mixture of peptides from non-structural proteins of hepatitis C virus. The greatest stimulating effect on T lymphocytes was exerted by DCs activated by nanoparticles, consisting of the recombinant mosaic protein with SAP at the N terminus and CTL epitopes of NS3, NS4a, and NS4b proteins at the C terminus. The presence of SAP in recombinant proteins’ sequences increased their immunogenicity. glycine linker at the BamH1-Sac1 to improve the interaction of the hexahistidine site with the Ni sorbent during purification. Nucleotide sequences optimized for E. coli encoded the recombinant proteins.