Chimeric HIV-1 virus-like particles containing gp120 epitopes as a result of a ribosomal frameshift elicit Gag- and SU-specific murine cytotoxic T-lymphocyte activities.
Insect cell expression of the HIV-1 Gag precursor protein by recombinant baculoviruses results in the assembly and budding of noninfectious virus-like particles (VLPs). The VLPs resemble immature virus in ultrastructural morphology and can be purified by conventional retroviral techniques. The virus-like appearance of the particles suggested that they could be used to package additional peptides. The retroviral frameshift mechanism was used to translate the pol gene products by expressing additional genetic information as chimeric Gag-Pol fusion proteins. Sequences encoding the carboxyl 65% of the HIV-1 surface glycoprotein (gp120, SU) were inserted into the Gag-Pol reading frame immediately downstream of the Gag stop codon. The assembly and budding of large quantities of Gag and chimeric Gag-SU VLPs were observed by standard transmission electron microscopy. The presence of gp120 epitopes in the Gag-SU VLPs was confirmed by immunoelectron microscopy and Western blot analysis using monoclonal anti-gp120 antibodies. Mice inoculated with the Gag-SU pseudovirions developed cytotoxic lymphocyte responses to both HIV-1 Gag and Env epitopes yet humoral immune responses only to Gag epitopes. The chimeric Gag-SU particles may have applications as vaccines or immunotherapeutic treatments for HIV-1 infection. In addition, the frameshift mechanism can be applied to the packaging of other viral or cellular proteins.
Tobin
G J
GJ
Laboratory of Cell and Molecular Structure, SAIC Frederick, Frederick Cancer Research and Development Center, Maryland 21701, USA. tobin@mail.ncifcrf.gov
Li
G H
GH
Fong
S E
SE
Nagashima
K
K
Gonda
M A
MA
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D002678:Chimera / Q000235:genetics / Q000276:immunology*; D017931:DNA Primers / Q000235:genetics; D000939:Epitopes / Q000235:genetics; D005260:Female; D016368:Frameshift Mutation; D015686:Gene Products, env / Q000235:genetics / Q000276:immunology; D015683:Gene Products, gag / Q000235:genetics / Q000276:immunology; D015699:HIV Envelope Protein gp120 / Q000235:genetics / Q000276:immunology*; D015497:HIV-1 / Q000235:genetics / Q000254:growth & development / Q000276:immunology*; D006801:Humans; D066298:In Vitro Techniques; D007181:Inclusion Bodies, Viral / Q000276:immunology / Q000648:ultrastructure / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D016253:Microscopy, Immunoelectron; D018411:Spodoptera; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
D017931:DNA Primers; D000939:Epitopes; D015686:Gene Products, env; D015683:Gene Products, gag; D015699:HIV Envelope Protein gp120