Enhanced exposure of the CD4-binding site to neutralizing antibodies by structural design of a membrane-anchored human immunodeficiency virus type 1 gp120 domain

Lan Wu, Tongqing Zhou, Zhi Yong Yang, Krisha Svehla, Sijy O'Dell, Mark K. Louder, Ling Xu, John R. Mascola, Dennis R. Burton, James A. Hoxie, Robert W. Doms, Peter D. Kwong, Gary J. Nabel

Research output: Contribution to journalArticle

Abstract

The broadly neutralizing antibody immunoglobulin G1 (IgG1) b12 binds to a conformationally conserved surface on the outer domain of the human immunodeficiency virus type 1 (HIV-1) gp120 envelope (Env) glycoprotein. To develop outer domain proteins (ODs) that could be recognized selectively by CD4-binding-site (CD4-BS) antibodies, membrane-anchored ODs were generated from an HIV-1 clade B virus, TA1 R3A, which was highly sensitive to neutralization by the IgG1 b12 antibody. A 231-residue fragment of gp120 (residues 252 to 482) linked to transmembrane regions from CD4 showed b12 binding comparable to that of the native Env spike as measured by flow cytometry. Truncation of the β20-β21 hairpin (residues 422 to 436 to Gly-Gly) improved overall protein expression. Replacement of the immunodominant central 20 amino acids of the V3 loop (residues 302 to 323) with a basic hexapeptide (NTRGRR) increased b12 reactivity further. Surface calculations indicated that the ratio of b12 epitope to exposed immunogenic surface in the optimized OD increased to over 30%. This OD variant [OD(GSL)(Δα20-21)(hCD4-TM)] was recognized by b12 and another CD4-BS-reactive antibody, b13, but not by eight other CD4-BS antibodies with limited neutralization potency. Furthermore, optimized membrane-anchored OD selectively absorbed neutralizing activity from complex antisera and b12. Structurally designed membrane-anchored ODs represent candidate immunogens to elicit or to allow the detection of broadly neutralizing antibodies to the conserved site of CD4 binding on HIV-1 gp120.

Original languageEnglish (US)
Pages (from-to)5077-5086
Number of pages10
JournalJournal of Virology
Volume83
Issue number10
DOIs
StatePublished - May 2009
Externally publishedYes

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Antibody Binding Sites
Human immunodeficiency virus 1
Neutralizing Antibodies
neutralizing antibodies
neutralization
HIV-1
binding sites
Binding Sites
immunoglobulins
Membranes
Immunoglobulins
Macacine herpesvirus 1
Cercopithecine Herpesvirus 1
antibodies
epitopes
flow cytometry
antiserum
Immune Sera
glycoproteins
Epitopes

ASJC Scopus subject areas

  • Immunology
  • Virology

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Enhanced exposure of the CD4-binding site to neutralizing antibodies by structural design of a membrane-anchored human immunodeficiency virus type 1 gp120 domain. / Wu, Lan; Zhou, Tongqing; Yang, Zhi Yong; Svehla, Krisha; O'Dell, Sijy; Louder, Mark K.; Xu, Ling; Mascola, John R.; Burton, Dennis R.; Hoxie, James A.; Doms, Robert W.; Kwong, Peter D.; Nabel, Gary J.

In: Journal of Virology, Vol. 83, No. 10, 05.2009, p. 5077-5086.

Research output: Contribution to journalArticle

Wu, L, Zhou, T, Yang, ZY, Svehla, K, O'Dell, S, Louder, MK, Xu, L, Mascola, JR, Burton, DR, Hoxie, JA, Doms, RW, Kwong, PD & Nabel, GJ 2009, 'Enhanced exposure of the CD4-binding site to neutralizing antibodies by structural design of a membrane-anchored human immunodeficiency virus type 1 gp120 domain', Journal of Virology, vol. 83, no. 10, pp. 5077-5086. https://doi.org/10.1128/JVI.02600-08
Wu, Lan ; Zhou, Tongqing ; Yang, Zhi Yong ; Svehla, Krisha ; O'Dell, Sijy ; Louder, Mark K. ; Xu, Ling ; Mascola, John R. ; Burton, Dennis R. ; Hoxie, James A. ; Doms, Robert W. ; Kwong, Peter D. ; Nabel, Gary J. / Enhanced exposure of the CD4-binding site to neutralizing antibodies by structural design of a membrane-anchored human immunodeficiency virus type 1 gp120 domain. In: Journal of Virology. 2009 ; Vol. 83, No. 10. pp. 5077-5086.
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abstract = "The broadly neutralizing antibody immunoglobulin G1 (IgG1) b12 binds to a conformationally conserved surface on the outer domain of the human immunodeficiency virus type 1 (HIV-1) gp120 envelope (Env) glycoprotein. To develop outer domain proteins (ODs) that could be recognized selectively by CD4-binding-site (CD4-BS) antibodies, membrane-anchored ODs were generated from an HIV-1 clade B virus, TA1 R3A, which was highly sensitive to neutralization by the IgG1 b12 antibody. A 231-residue fragment of gp120 (residues 252 to 482) linked to transmembrane regions from CD4 showed b12 binding comparable to that of the native Env spike as measured by flow cytometry. Truncation of the β20-β21 hairpin (residues 422 to 436 to Gly-Gly) improved overall protein expression. Replacement of the immunodominant central 20 amino acids of the V3 loop (residues 302 to 323) with a basic hexapeptide (NTRGRR) increased b12 reactivity further. Surface calculations indicated that the ratio of b12 epitope to exposed immunogenic surface in the optimized OD increased to over 30{\%}. This OD variant [OD(GSL)(Δα20-21)(hCD4-TM)] was recognized by b12 and another CD4-BS-reactive antibody, b13, but not by eight other CD4-BS antibodies with limited neutralization potency. Furthermore, optimized membrane-anchored OD selectively absorbed neutralizing activity from complex antisera and b12. Structurally designed membrane-anchored ODs represent candidate immunogens to elicit or to allow the detection of broadly neutralizing antibodies to the conserved site of CD4 binding on HIV-1 gp120.",
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AU - O'Dell, Sijy

AU - Louder, Mark K.

AU - Xu, Ling

AU - Mascola, John R.

AU - Burton, Dennis R.

AU - Hoxie, James A.

AU - Doms, Robert W.

AU - Kwong, Peter D.

AU - Nabel, Gary J.

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