Abstract
Objective:
To develop an HIV-1 accessory gene immunogen using a DNA vaccine approach.
Methods:
HIV-1 accessory genes vif, vpu and nef were modified to express under the control of a single promoter with cellular proteolytic cleavage sites between the coding sequences (VVN-P). Immune responses induced by these constructs were evaluated in mice.
Results:
DNA vaccine construct (pVVN-P) expressing Vif, Vpu and Nef was processed and the fusion protein was cleaved appropriately. Vif, Vpu and Nef as a fusion protein with proteolytic cleavage sites (VVN-P) is able to induce a significant level of cellular immune responses. We also observed that accessory genes Vif, Vpu and Nef (VVN-P) induced an effective T helper 1 proliferative response measured by cytokine production. Furthermore, expression cassette pVVN-P was able to induce cytotoxic T lymphocyte (CTL) responses against diverse HIV-1 viruses in infected target cells.
Conclusion:
We conclude that cell-mediated immune responses induced by accessory gene constructs from clade B may have a broader recognition of divergent HIV-1 viruses and should be further examined for both prophylactic and therapeutic vaccination schemes against HIV-1.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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3T3 Cells
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Animals
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Blotting, Western
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Cell Division / immunology
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Cells, Cultured
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Cytokines / biosynthesis
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Cytotoxicity Tests, Immunologic
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Fluorescent Antibody Technique
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Gene Products, nef / genetics
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Gene Products, nef / immunology
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Gene Products, nef / metabolism
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Gene Products, vif / genetics
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Gene Products, vif / immunology
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Gene Products, vif / metabolism
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HIV-1 / genetics
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HIV-1 / immunology*
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HIV-1 / isolation & purification
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HeLa Cells
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Human Immunodeficiency Virus Proteins
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Humans
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Injections, Intramuscular
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Mice
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Mice, Inbred BALB C
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / immunology
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Recombinant Fusion Proteins / metabolism
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T-Lymphocytes, Helper-Inducer / cytology
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T-Lymphocytes, Helper-Inducer / metabolism
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Transfection
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Vaccination
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Vaccines, DNA / genetics
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Vaccines, DNA / immunology*
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Vaccines, DNA / metabolism
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Viral Regulatory and Accessory Proteins / genetics
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Viral Regulatory and Accessory Proteins / immunology
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Viral Regulatory and Accessory Proteins / metabolism
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nef Gene Products, Human Immunodeficiency Virus
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vif Gene Products, Human Immunodeficiency Virus
Substances
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Cytokines
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Gene Products, nef
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Gene Products, vif
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Human Immunodeficiency Virus Proteins
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Recombinant Fusion Proteins
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Vaccines, DNA
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Viral Regulatory and Accessory Proteins
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nef Gene Products, Human Immunodeficiency Virus
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vif Gene Products, Human Immunodeficiency Virus
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vpu protein, Human immunodeficiency virus 1