Assembly and immunogenicity of chimeric Gag-Env proteins derived from the human immunodeficiency virus type 1.
We evaluated the potential of the precursor Gag protein (Pr55) of the human immunodeficiency virus type 1 (HIV-1) as a carrier for the presentation of envelope epitopes. Recombinant chimeric core-envelope protein-expressing constructs were derived by deletion of regions within the gag gene, especially of regions encoding p24 capsid epitopes. Sequences encoding either the principal neutralization determinant (PND) and/or the CD4-binding domains (CD4BS) were then inserted. Deletion of residues 196-226 within the p24 capsid protein did not prevent self-assembly into virus-like particles (VLPs) whereas deletion of residues 299-328 completely abolished VLP formation. Thus the major homology region (MHR) and proximal sequences are required for capsid assembly. An immunization study in mice showed that assembled chimeric proteins elicited strong anti-Gag, weak anti-envelope, and no neutralizing humoral responses. Nonassembled chimeric proteins were poor immunogens. Mapping of Pr55 antigenic sites using sera from immunized mice and peptides overlapping the entire Gag precursor showed that p24 capsid and p17 matrix epitopes presented to the immune system differed from the mature form (p24 or p17) and the multimeric immature form (Pr55).
Laboratoire de Virologie, Centre National de la Recherche Scienifique, Centre Hospitalier Universitaire Bretonneau, France.
C
C
Truong
D
D
Brand
F
F
Mallet
P
P
Roingeard
S
S
Brunet
F
F
Barin
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018604:Epitope Mapping; D005260:Female; D017353:Gene Deletion; D015686:Gene Products, env / Q000235:genetics* / Q000276:immunology; D015683:Gene Products, gag / Q000235:genetics* / Q000276:immunology; D015497:HIV-1 / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology*; D017421:Sequence Analysis
D015686:Gene Products, env; D015683:Gene Products, gag; D011993:Recombinant Fusion Proteins