PloS one, 2013-05, Vol.8 (5), p.e62720
2013
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- Autor(en) / Beteiligte
- Titel
- Consensus HIV-1 FSU-A integrase gene variants electroporated into mice induce polyfunctional antigen-specific CD4+ and CD8+ T cells
- Ist Teil von
- PloS one, 2013-05, Vol.8 (5), p.e62720
- Ort / Verlag
- United States: Public Library of Science
- Erscheinungsjahr
- 2013
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- Our objective is to create gene immunogens targeted against drug-resistant HIV-1, focusing on HIV-1 enzymes as critical components in viral replication and drug resistance. Consensus-based gene vaccines are specifically fit for variable pathogens such as HIV-1 and have many advantages over viral genes and their expression-optimized variants. With this in mind, we designed the consensus integrase (IN) of the HIV-1 clade A strain predominant in the territory of the former Soviet Union and its inactivated derivative with and without mutations conferring resistance to elvitegravir. Humanized IN gene was synthesized; and inactivated derivatives (with 64D in the active site mutated to V) with and without elvitegravir-resistance mutations were generated by site-mutagenesis. Activity tests of IN variants expressed in E coli showed the consensus IN to be active, while both D64V-variants were devoid of specific activities. IN genes cloned in the DNA-immunization vector pVax1 (pVaxIN plasmids) were highly expressed in human and murine cell lines (>0.7 ng/cell). Injection of BALB/c mice with pVaxIN plasmids followed by electroporation generated potent IFN-γ and IL-2 responses registered in PBMC by day 15 and in splenocytes by day 23 after immunization. Multiparametric FACS demonstrated that CD8+ and CD4+ T cells of gene-immunized mice stimulated with IN-derived peptides secreted IFN-γ, IL-2, and TNF-α. The multi-cytokine responses of CD8+ and CD4+ T-cells correlated with the loss of in vivo activity of the luciferase reporter gene co-delivered with pVaxIN plasmids. This indicated the capacity of IN-specific CD4+ and CD8+ T-cells to clear IN/reporter co-expressing cells from the injection sites. Thus, the synthetic HIV-1 clade A integrase genes acted as potent immunogens generating polyfunctional Th1-type CD4+ and CD8+ T cells. Generation of such response is highly desirable for an effective HIV-1 vaccine as it offers a possibility to attack virus-infected cells via both MHC class I and II pathways.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1932-6203eISSN: 1932-6203DOI: 10.1371/journal.pone.0062720Titel-ID: cdi_plos_journals_1350492313
- Format
- –
- Schlagworte
- Animals, Antigens, Biology, CD4 antigen, CD4-Positive T-Lymphocytes - immunology, CD8 antigen, CD8-Positive T-Lymphocytes - immunology, Cell Line, Cell lines, Clinical trials, Critical components, Deoxyribonucleic acid, DNA, DNA vaccines, Drug resistance, Drug Resistance, Viral - genetics, Drug Resistance, Viral - immunology, E coli, Electroporation, Enzymes, Escherichia coli, Flow Cytometry, Gene expression, Genes, Genetic engineering, Genetic vectors, Genomes, HIV, HIV Integrase - biosynthesis, HIV Integrase - genetics, HIV Integrase Inhibitors - metabolism, HIV-1 - enzymology, HIV-1 - immunology, House mouse, Human immunodeficiency virus, Humans, Immunization, In vivo methods and tests, Infections, Influenza, Injection, Integrase, Interferon, Interleukin 2, Luciferases, Lymphocyte Activation - immunology, Lymphocytes, Lymphocytes T, Major histocompatibility complex, Medicin och hälsovetenskap, Medicine, Mice, Mice, Inbred BALB C, Microbial drug resistance, Molecular biology, Mutagenesis, Mutagenesis, Site-Directed, Mutation, Peptides, Peripheral blood mononuclear cells, Plasmids, Quinolones, Reporter gene, Splenocytes, T cells, Territory, Tumor necrosis factor-α, Vaccines, Viral infections, Virology, Viruses, γ-Interferon