Expression of human immunodeficiency virus type 1 Gag protein precursor and envelope proteins from a vesicular stomatitis virus recombinant: high-level production of virus-like particles containing HIV envelope.
Recombinant vesicular stomatitis viruses have been developed as high-level expression vectors which serve as effective vaccine vectors in animals (Roberts et al., 1998, J. Virol. 72, 4704-4711; Roberts et al., 1999, J. Virol. 73, 3723-3732). Here we show that two genes can be expressed simultaneously from a single, live-attenuated VSV recombinant. The genes used encode the Pr55(gag) protein precursor of HIV-1 (1.7-kb gene) and an HIV-1 envelope (Env) protein (2.4 kb gene). Our results show that VSV can accommodate up to a 40% increase in genome size with only a threefold reduction in virus titer. Recombinants expressing the Pr55(gag) protein precursor with or without Env protein produced abundant HIV virus-like particles (VLPs) in addition to bullet-shaped VSV particles. HIV Env protein expressed from a VSV recombinant also expressing Gag was specifically incorporated into the HIV VLPs but not into the VSV particles. In contrast, VSV G protein was found in both VSV particles and in HIV VLPs. Such VSV/HIV recombinants producing HIV VLPs with Env protein could be an effective source of HIV-like particles inducing both cellular and antibody-mediated immunity to HIV-1.
Haglund
K
K
Department of Pathology, Yale University School of Medicine, New Haven, Connecticut, 06510, USA.
Forman
J
J
Kräusslich
H G
HG
Rose
J K
JK
D016915:AIDS Vaccines / Q000235:genetics; D000818:Animals; D002460:Cell Line; D004274:DNA, Recombinant / Q000235:genetics; D015683:Gene Products, gag / Q000235:genetics / Q000378:metabolism*; D005822:Genetic Vectors; D015699:HIV Envelope Protein gp120 / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000235:genetics* / Q000502:physiology*; D006801:Humans; D008562:Membrane Glycoproteins; D008854:Microscopy, Electron; D010957:Plasmids / Q000235:genetics; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014613:Vaccines, Attenuated / Q000235:genetics / Q000378:metabolism; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D014759:Viral Envelope Proteins / Q000235:genetics; D014771:Virion / Q000378:metabolism / Q000502:physiology
D016915:AIDS Vaccines; D004274:DNA, Recombinant; C028391:G protein, vesicular stomatitis virus; D015683:Gene Products, gag; D015699:HIV Envelope Protein gp120; D008562:Membrane Glycoproteins; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; D014613:Vaccines, Attenuated; D014759:Viral Envelope Proteins; C059767:p55 gag precursor protein, Human immunodeficiency virus 1