Endotoxin-minimized HIV-1 p24 fused to murine hsp70 activates dendritic cells, facilitates endocytosis and p24-specific Th1 response in mice
Language English Country Netherlands Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
26021827
DOI
10.1016/j.imlet.2015.05.010
PII: S0165-2478(15)00086-3
Knihovny.cz E-resources
- Keywords
- HIV, HIV-1 protein p24, Heat shock protein, Vaccine,
- MeSH
- Antigens, Viral immunology MeSH
- CD8-Positive T-Lymphocytes immunology MeSH
- Dendritic Cells immunology MeSH
- Endocytosis immunology MeSH
- Endotoxins immunology MeSH
- HIV Core Protein p24 immunology metabolism MeSH
- Immunization MeSH
- Mice, Inbred BALB C MeSH
- Mice MeSH
- Cross-Priming immunology MeSH
- HSP70 Heat-Shock Proteins immunology metabolism MeSH
- Recombinant Fusion Proteins immunology MeSH
- Th1 Cells immunology MeSH
- AIDS Vaccines immunology MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Antigens, Viral MeSH
- Endotoxins MeSH
- HIV Core Protein p24 MeSH
- HSP70 Heat-Shock Proteins MeSH
- Recombinant Fusion Proteins MeSH
- AIDS Vaccines MeSH
Heat shock proteins hsp70 and gp96 have been confirmed as adjuvants enabling induction of cell- and antibody-mediated immunity specific to associated protein or peptide antigens due to the activation of naive dendritic cells and supporting cross-presentation of associated antigen. An efficacious vaccine preventing HIV-1 infection should induce (1) antibodies neutralizing HIV-1 Env protein, preventing virus spreading and (2) CD4(+) Th1 and CD8(+) T cells specific to viral proteins, especially gag p24, important for elimination of HIV-1 infected cells. As p24 is relatively poorly recognized by dendritic cells, its targeting to DC is important for enhancement of vaccine efficacy. In this study, a p24 protein fused to the C- or N-terminus of murine hsp70 was produced as a recombinant protein and administered without any adjuvant to experimental BALB/c mice. Consequently, p24-specific cellular and humoral immune responses were measured. To minimize the effect of bacterial endotoxin, each protein was subjected to a repeated endotoxin phase extraction until each preparation contained less than 2.5 endotoxin unit (EU) per mg of antigen. In addition, endocytosis of p24 fused to hsp70 by dendritic cells and their activation were characterized. The fusion to hsp70 protein enhanced endocytosis of p24 as well as activation of dendritic cells in vitro. After immunization of mice, hsp70-p24 fusion protein induced the strongest p24-specific CD4(+) and CD8(+) T cells (IFN-γ production) and humoral (IgG2b) responses corresponding to Th1 type dominance, whereas p24-hsp70 or p24 itself induced weaker responses.
References provided by Crossref.org
