title: Selective recognition of oligomeric HIV-1 primary isolate envelope glycoproteins by potently neutralizing ligands requires efficient precursor cleavage.
abstract: A critical component of an effective HIV vaccine will be the induction of broadly neutralizing antibodies. Comprising the HIV spike, the exterior envelope glycoprotein gp120 and the transmembrane glycoprotein gp41 mediate receptor binding, viral entry, and are the targets for neutralizing antibodies. The gp120 and gp41 glycoproteins are derived from the gp160 precursor glycoprotein and following gp160 glycosylation, oligomerization and cleavage in the endoplasmic reticulum and Golgi, remain as non-covalently associated trimers of heterodimers. Previously, using cell-surface envelope glycoproteins derived from infection of a laboratory-adapted HIV-1 strain, a correlation had been established between the binding of gp120-directed antibodies to the viral glycoprotein and the ability of the antibodies to neutralize laboratory-adapted isolates. However, this has been more difficult to demonstrate for glycoproteins derived from primary patient isolates. Here, using a FACS-based method, we report that only gp120-directed neutralizing antibodies and the neutralizing ligand soluble CD4 efficiently bind to glycoproteins derived from the JR-FL primary isolate provided that the gp160 precursor protein is efficiently cleaved. Precursor cleavage was demonstrated by cell-surface biotinylation and Western blotting. In stark contrast, both non-neutralizing and neutralizing antibodies bind non-cleaved envelope glycoproteins from JR-FL and YU2 isolates. These data imply that significant changes in Env spike structure are dependent upon precursor gp160 cleavage and are consistent with a restricted-binding-to-Env model of neutralization. The data also have implications in regards to the use and design of non-cleaved envelope glycoprotein trimeric immunogens as a means to selectively and preferentially present neutralizing epitopes to the host immune system.
authors.affiliation: Structural Virology Section, Vaccine Research Center, NIAID, National Institutes of Health, 40 Convent Drive, Building 40, Room 4512, Bethesda, MD 20892, USA.
authors.forename: Marie
authors.initials: M
authors.lastname: Pancera
authors.forename: Richard
authors.initials: R
authors.lastname: Wyatt
mesh_terms: D000956:Antigens, Viral / Q000096:biosynthesis / Q000737:chemistry*; D002460:Cell Line; D006023:Glycoproteins / Q000737:chemistry* / Q000235:genetics; D015483:HIV Antibodies / Q000097:blood*; D019302:HIV Envelope Protein gp160 / Q000032:analysis; D015497:HIV-1 / Q000737:chemistry* / Q000276:immunology; D008024:Ligands; D009500:Neutralization Tests; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology
chemical_list: D000956:Antigens, Viral; D006023:Glycoproteins; D015483:HIV Antibodies; D019302:HIV Envelope Protein gp160; D008024:Ligands; D011993:Recombinant Fusion Proteins