The structural HCV genes delivered by MPG cell penetrating peptide are directed to enhance immune responses in mice model

Abstract

One of the significant problems in vaccination projects is the lack of an effective vaccine against hepatitis C virus (HCV). The goal of the current study is to evaluate and compare two DNA constructs encoding HCV core and coreE1E2 genes alone or complexed with MPG peptide as a delivery system for stimulation of antibody responses and IFN-γ secretion in Balb/c mice model. Indeed, MPG cell penetrating peptide was used to improve DNA immunization in mice. Our results demonstrated that MPG forms stable non-covalent nanoparticles with pcDNA-core and pcDNA-coreE1E2 at an N/P ratio of 10:1. The in vitro transfection efficiency of core or coreE1E2 DNA using MPG and TurboFect delivery systems was confirmed by western blot analysis. The results indicated the expression of the full-length core (∼21 kDa), and coreE1E2 (∼83 kDa) proteins using an anti-His monoclonal antibody. In addition, the expression of HCV core and coreE1E2 proteins was performed in bacteria and the purified recombinant proteins were injected to mice with Montanide 720 adjuvant. Our data showed that the immunized mice with HCV core and coreE1E2 proteins generated the mixture of sera IgG1 and IgG2a isotypes considerably higher than other groups. Furthermore, DNA constructs encoding core and coreE1E2 complexed with MPG could significantly induce IFN-γ secretion in lower concentrations than the naked core and coreE1E2 DNAs. Taken together, the DNA formulations as well as protein regimens used in this study triggered high-level IFN-γ production in mice, an important feature for the development of Th1 immune responses.

Cell penetrating peptides: the potent multi-cargo intracellular carriers

“…In this line, HPV16 E7 DNA/MPG in nanoparticle formulation at an N/P ratio of 10:1 elicited an effective Th1 cellular immune response and completely protected mice against tumors, as well [264]. In another study, the induction of cell-mediated immune response (Th1-biased response) against HCV core and HCV coreE1E2 antigens was stronger in mice immunized with coreE1E2 DNA/MPG and then core DNA/MPG nanoparticles as compared to coreE1E2 and core DNA constructs alone [265]. In addition, the use of Tat CPP fused to Nef antigen (Tat-Nef) in heterologous prime-boost strategy along with Cady-2 CPP significantly induced the Nef-specific T cell responses for development of HIV-1 vaccine [266].…”

Use of Protamine in Nanopharmaceuticals—A Review

“…In this case, two DNA constructs encoding HCV core and coreE1E2 genes were complexed with MPG, and then the efficacy of the complexes was compared to that of the antigens used alone in Balb/c mice. Mice immunized with the complexes generated a mixture of IgG1 and IgG2 antibodies as well as increased IFN-γ production [265]. Furthermore, MPG was used to assist antigen cross-presentation and increase the tumor immune response to a tumor vaccine.…”

Peptide-Based Nanoparticles for Therapeutic Nucleic Acid Delivery

“…Since 2015, only a few papers were published using MPG as a unique oligonucleotide delivery system, and this peptide was used more as a grafted entity on PLGA polymers in order to increase their cellular translocation [101,102]. However, some recent works were recently published from Dr. A. Bolhassani's group using MPG alone for the delivery of genes to develop an effective vaccine against the hepatitis C virus (HCV) [103] or human immunodeficiency virus (HIV) [104].…”