Simultaneous use of natural adjuvants and cell penetrating peptides improves HCV NS3 antigen-specific immune responses
| Autor: | Seyed Mehdi Sadat, Shiva Irani, Sina Alizadeh, Azam Bolhassani |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Viral Hepatitis Vaccines
0301 basic medicine Recombinant Fusion Proteins viruses medicine.medical_treatment Immunology HSP27 Heat-Shock Proteins Cell-Penetrating Peptides Hepacivirus Viral Nonstructural Proteins Mice 03 medical and health sciences Immunogenicity Vaccine 0302 clinical medicine Immune system Adjuvants Immunologic Antigen Heat shock protein Vaccines DNA medicine Animals Humans Immunology and Allergy NS3 biology Chemistry virus diseases biochemical phenomena metabolism and nutrition Hepatitis C Molecular biology digestive system diseases Granzyme B Disease Models Animal HEK293 Cells 030104 developmental biology Cytokine biology.protein Female Antibody Adjuvant 030215 immunology |
| Zdroj: | Immunology Letters. 212:70-80 |
| ISSN: | 0165-2478 |
| Popis: | To improve an effective hepatitis C virus (HCV) therapeutic vaccine, induction of a strong and long term HCV antigen-specific immune response is an important parameter. HCV non-structural protein 3 (NS3) has antigenic properties and plays a major role in viral clearance. In this study, DNA constructs encoding HCV NS3 and heat shock protein 27 (Hsp27)-NS3 genes, and the recombinant (r) NS3 and rHsp27-NS3 proteins complexed with HR9 and Cady-2 cell penetrating peptides (CPPs) were utilized to evaluate antibody, cytokine and Granzyme B secretion in mice. Herein, the formation of NS3 and Hsp27-NS3 DNA/ HR9 CPP complexes were revealed by gel retardation assay and protection against DNase and protease. Cady-2 peptide was used to form the nanoparticles with rNS3 and rHsp27-NS3 proteins. The size and charge of the nanoparticles were confirmed by SEM and Zetasizer instruments. Next, in vitro transfection of the nanoparticles was assessed by flow cytometry and western blotting. Finally, humoral and cellular immune responses were evaluated using different modalities in mice. Our data showed that HR9 and Cady-2 could form stable nanoparticles with DNA and proteins, respectively and enhance their delivery into HEK-293 T cells in a non-covalent approach. Furthermore, the heterologous Hsp27-NS3 DNA + HR9 prime/rHsp27-NS3+Cady-2 protein boost elicited a higher Th1 cellular immune response with a predominant IgG2a, IgG2b, IFN-γ profile and strong Granzyme B secretion than those induced by other groups. Briefly, the combination of a natural adjuvant (Hsp27) and CPPs (HR9 and Cady-2) could significantly stimulate effective immune responses as a promising approach for development of HCV therapeutic vaccines. |
| Databáze: | OpenAIRE |
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