Improved antigenicity of the HIV env protein by cleavage site removal.
The HIV env glycoprotein mediates virus infection and cell fusion through an interaction with the CD4 molecule present at the surface of T4+ lymphocytes. Although env presents a major antigenic target, vaccinia recombinants expressing env elicit low titres of anti-env antibody (Kieny et al., Bio/Technology, 4, 790-795, 1986). To delimit the functional domains of env and to improve the immunogenicity of the vaccinia recombinants we constructed variants expressing env proteins in which the site permitting cleavage of the gp160 precursor to yield gp120 and gp41 was removed, the gp120 and gp41 moieties separated or in which the signal sequence and hydrophobic domains were replaced by equivalents from rabies virus G. Analysis of variants revealed that the gp120 moiety is alone capable of interacting with CD4 and of provoking aggregation of T4+ lymphocytes, whereas cell-associated gp41 liberated by gp160 cleavage was essential for cell fusion. The identity of the signal and transmembrane zones however appeared unimportant. Although removal of the consensus sequence permitting cleavage of gp160 prevented syncytium formation but not aggregation of T4+ lymphocytes, significant cleavage continued to take place. Removal of a second potential cleavage site blocked gp160 cleavage. The live viruses were examined for immunogenicity: recombinant 1139 which lacks both putative cleavage sites was found to elicit a 10-fold higher antibody response in experimental animals than the parental recombinant.
Kieny
M P
MP
Transgène S.A., Strasbourg, France.
Lathe
R
R
Rivière
Y
Y
Dott
K
K
Schmitt
D
D
Girard
M
M
Montagnier
L
L
Lecocq
J
J
D000595:Amino Acid Sequence; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D006678:HIV / Q000472:pathogenicity*; D015488:HIV Antigens / Q000276:immunology*; D019302:HIV Envelope Protein gp160; D006868:Hydrolysis; D008969:Molecular Sequence Data; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000096:biosynthesis; D012191:Retroviridae Proteins / Q000276:immunology*; D014759:Viral Envelope Proteins / Q000276:immunology*
D015488:HIV Antigens; D019302:HIV Envelope Protein gp160; D011994:Recombinant Proteins; D012191:Retroviridae Proteins; D014759:Viral Envelope Proteins