Characterization of a DNA vaccine expressing a human immunodeficiency virus-like particle

An ideal human immunodeficiency virus type-1 (HIV-1) vaccine will most likely need to elicit cross-reactive neutralizing antibodies and a strong cell-mediated immune response against multiple HIV-1 antigens to confer protection against challenge. In this study, DNA vaccines were constructed to express virally regulated human immunodeficiency virus-like particles (VLP) to elicit broad-spectrum immune responses to multiple HIV-1 antigens. VLPs were efficiently produced using sequences encoding gag and pol gene products from an X4 isolate and sequences encoding for tat, rev, vpu, and env from R5 or R5X4 isolates. The integrase, vpr, vif, and nef genes were deleted. In addition, the long terminal repeats (LTRs) were removed and transcription of the VLP insert was driven by the addition of the cytomegalovirus immediate-early (CMV-IE) promoter. A second generation of VLP vaccine plasmids was constructed with mutations engineered into the VLP DNA to produce particles deficient in activities associated with viral reverse transcriptase and protease. Primate cell lines, transiently transfected with DNA, efficiently secreted VLP into the supernatant that banded within a sucrose gradient at densities similar to infectious virions. In addition, these particles incorporated Env on the particle surface that bound soluble human CD4. These VLPs provide a safe and efficient strategy for presenting multiple HIV-1 antigens, expressed from a single insert, to the immune system in a structure that mimics the infectious virion.