Immunogenicity of a novel DNA vaccine cassette expressing multiple human immunodeficiency virus (HIV-1) accessory genes.
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.


CONCLUSIONS
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.
Ayyavoo
V
V
Department of Pathology & Laboratory Medicine, University of Pennsylvania, Philadelphia 19104, USA.
Kudchodkar
S
S
Ramanathan
M P
MP
Le
P
P
Muthumani
K
K
Megalai
N M
NM
Dentchev
T
T
Santiago-Barrios
L
L
Mrinalini
C
C
Weiner
D B
DB
D016475:3T3 Cells; D000818:Animals; D015153:Blotting, Western; D002455:Cell Division / Q000276:immunology; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis; D003601:Cytotoxicity Tests, Immunologic; D005455:Fluorescent Antibody Technique; D015693:Gene Products, nef / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D016346:Gene Products, vif / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D006367:HeLa Cells; D054298:Human Immunodeficiency Virus Proteins; D006801:Humans; D007273:Injections, Intramuscular; D051379:Mice; D008807:Mice, Inbred BALB C; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006377:T-Lymphocytes, Helper-Inducer / Q000166:cytology / Q000378:metabolism; D014162:Transfection; D014611:Vaccination; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D054334:Viral Regulatory and Accessory Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D054311:nef Gene Products, Human Immunodeficiency Virus; D054320:vif Gene Products, Human Immunodeficiency Virus; D000086082:Viroporin Proteins
D016207:Cytokines; D015693:Gene Products, nef; D016346:Gene Products, vif; D054298:Human Immunodeficiency Virus Proteins; D011993:Recombinant Fusion Proteins; D019444:Vaccines, DNA; D054334:Viral Regulatory and Accessory Proteins; D054311:nef Gene Products, Human Immunodeficiency Virus; D054320:vif Gene Products, Human Immunodeficiency Virus; C514859:vpu protein, Human immunodeficiency virus 1; D000086082:Viroporin Proteins