Structural Basis of the Cross-Reactivity of Genetically Related Human Anti-HIV-1 mAbs: Implications for Design of V3-Based Immunogens

Burke, Valicia; Williams, Constance; Sukumaran, Madhav; Kim, Seung-Sup; Li, Huiguang; Wang, Xiaohong; Gorny, Miroslaw K.; Zolla‐Pazner, Susan; Kong, Xiang‐Peng

doi:10.1016/j.str.2009.09.012

Cited by 79 publications

(112 citation statements)

References 56 publications

(86 reference statements)

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“…Signature motifs were derived previously based on the 3D shapes of epitopes in V3 that are recognized by human MAbs which are able to neutralize diverse strains of viruses from multiple clades (3,14,64). Briefly, by studying the crystal structures of MAb/epitope complexes (13,36,60,61), identifying the key V3 residues that are recognized by the MAb, and determining the stringency with which these residues are required for neutralization by that MAb, a signature motif for the epitope recognized by that MAb can be defined. For example, the signature motif for the relatively clade B-restricted anti-V3 MAb 447-52D is P 16 R 18 , where proline is required at position 16 in the V3 loop and arginine is required at position 18 (14,64).…”

Section: Resultsmentioning

confidence: 99%

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Cross-Clade HIV-1 Neutralizing Antibodies Induced with V3-Scaffold Protein Immunogens following Priming with gp120 DNA

Zolla‐Pazner

Kong

Jiang

et al. 2011

J Virol

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The V3 epitope is a known target for HIV-1 neutralizing antibodies (NAbs), and V3-scaffold fusion proteins used as boosting immunogens after gp120 DNA priming were previously shown to induce NAbs in rabbits. Here, we evaluated whether the breadth and potency of the NAb response could be improved when boosted with rationally designed V3-scaffold immunogens. Rabbits were primed with codon-optimized clade C gp120 DNA and boosted with one of five V3-cholera toxin B fusion proteins (V3-CTBs) or with double combinations of these. The inserts in these immunogens were designed to display V3 epitopes shared by the majority of global HIV-1 isolates. Double combinations of V3-CTB immunogens generally induced more broad and potent NAbs than did boosts with single V3-CTB immunogens, with the most potent and broad NAbs elicited with the V3-CTB carrying the consensus V3 of clade C (V3 C -CTB), or with double combinations of V3-CTB immunogens that included V3 C -CTB. Neutralization of tier 1 and 2 pseudoviruses from clades AG, B, and C and of peripheral blood mononuclear cell (PBMC)-grown primary viruses from clades A, AG, and B was achieved, demonstrating that priming with gp120 DNA followed by boosts with V3-scaffold immunogens effectively elicits cross-clade NAbs. Focusing on the V3 region is a first step in designing a vaccine targeting protective epitopes, a strategy with potential advantages over the use of Env, a molecule that evolved to protect the virus by poorly inducing NAbs and by shielding the epitopes that are most critical for infectivity.

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Section: Resultsmentioning

confidence: 99%

“…Recently, more detailed data have become available mapping the conserved structural elements in the V3 crown within and between HIV subtypes (3,13,14,36,71). Using these data, newly designed and optimally constructed V3-scaffold protein immunogens have now been synthesized and tested for antigenicity in vitro and for immunogenicity in vivo in rabbits.…”

mentioning

confidence: 99%

Cross-Clade HIV-1 Neutralizing Antibodies Induced with V3-Scaffold Protein Immunogens following Priming with gp120 DNA

Zolla‐Pazner

Kong

Jiang

et al. 2011

J Virol

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“…In such cradle-like Abs the antigen binding site consists of a groove in the Fab fragment, and the epitope lies in this groove, resembling a baby in a cradle; in this case, the major binding site is the hydrophobic core of the V3 crown, usually composed of hydrophobic, conserved residues 307, 309, and 317 (89,96,97).…”

Section: Glycan-independent "Ladle-like" V3 Absmentioning

confidence: 99%

Antibodies Targeting the Envelope of HIV-1

Mayr

Zolla‐Pazner²

2015

Microbiol Spectr

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Antibodies (Abs) are a critical component of the human immune response against viral infections. In HIV-infected patients, a robust Ab response against the virus develops within months of infection; however, due to numerous strategies, the virus usually escapes the biological effects of the various Abs. Here we provide an overview of the different viral evasion mechanisms, including glycosylation, high mutation rate, and conformational masking by the envelope glycoproteins of the virus. In response to virus infection and to its evolution within a host, "conventional Abs" are generated, and these can also be induced by immunization; generally, these Abs are limited in their neutralization breadth and potency. In contrast, "exceptional Abs" require extended exposure to virus to generate the required hypermutation in the immunoglobulin variable regions, and they occur only in rare HIV-infected individuals, but they display impressive breadth and potency. In this review, we describe the major regions of the HIV envelope spike that are targeted by conventional and exceptional Abs. These include the first, second, and third variable loops (V1, V2, and V3) located at the apex of the envelope trimer, the CD4 binding site, and the membrane-proximal external region of the gp41 ectodomain. Lastly, we discuss the challenging task of HIV immunogen design and approaches for choosing which immunogens might be used to elicit protective Abs.

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“…Model rebuilding was performed using Coot 37 and refinement was performed using autobuster 38 with LSSR restraints. 39 High resolution Fab fragment structures with high homology to Fpro0165 Fab were used to build a model for target restraints (PDB code 3AAZ 40 and PDB code 3GHB 41 for the light and heavy chains respectively). The structural coordinates of Fpro0165 Fab have been deposited in the Protein Data Bank (PDB code 4UV4).…”

Section: Crystallography and Structural Determination Of Fpro0165 Fabmentioning

confidence: 99%

Affinity maturation of a novel antagonistic human monoclonal antibody with a long V_HCDR3 targeting the Class A GPCR formyl-peptide receptor 1

Douthwaite

Sridharan

Huntington

et al. 2015

mAbs

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(2015) Affinity maturation of a novel antagonistic human monoclonal antibody with a long V H CDR3 targeting the Class A GPCR formyl-peptide receptor 1, mAbs, 7:1, 152-166, DOI: 10.4161/19420862.2014.985158 To link to this article: https://doi.org/10. 4161/19420862.2014 Therapeutic monoclonal antibodies targeting G-protein-coupled receptors (GPCRs) are desirable for intervention in a wide range of disease processes. The discovery of such antibodies is challenging due to a lack of stability of many GPCRs as purified proteins. We describe here the generation of Fpro0165, a human anti-formyl peptide receptor 1 (FPR1) antibody generated by variable domain engineering of an antibody derived by immunization of transgenic mice expressing human variable region genes. Antibody isolation and subsequent engineering of affinity, potency and species cross-reactivity using phage display were achieved using FPR1 expressed on HEK cells for immunization and selection, along with calcium release cellular assays for antibody screening. Fpro0165 shows full neutralization of formyl peptide-mediated activation of primary human neutrophils. A crystal structure of the Fpro0165 Fab shows a long, protruding V H CDR3 of 24 amino acids and in silico docking with a homology model of FPR1 suggests that this long V H CDR3 is critical to the predicted binding mode of the antibody. Antibody mutation studies identify the apex of the long V H CDR3 as key to mediating the species cross-reactivity profile of the antibody. This study illustrates an approach for antibody discovery and affinity engineering to typically intractable membrane proteins such as GPCRs.

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Structural Basis of the Cross-Reactivity of Genetically Related Human Anti-HIV-1 mAbs: Implications for Design of V3-Based Immunogens

Cited by 79 publications

References 56 publications

Cross-Clade HIV-1 Neutralizing Antibodies Induced with V3-Scaffold Protein Immunogens following Priming with gp120 DNA

Cross-Clade HIV-1 Neutralizing Antibodies Induced with V3-Scaffold Protein Immunogens following Priming with gp120 DNA

Antibodies Targeting the Envelope of HIV-1

Affinity maturation of a novel antagonistic human monoclonal antibody with a long V_HCDR3 targeting the Class A GPCR formyl-peptide receptor 1

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Structural Basis of the Cross-Reactivity of Genetically Related Human Anti-HIV-1 mAbs: Implications for Design of V3-Based Immunogens

Cited by 79 publications

References 56 publications

Cross-Clade HIV-1 Neutralizing Antibodies Induced with V3-Scaffold Protein Immunogens following Priming with gp120 DNA

Cross-Clade HIV-1 Neutralizing Antibodies Induced with V3-Scaffold Protein Immunogens following Priming with gp120 DNA

Antibodies Targeting the Envelope of HIV-1

Affinity maturation of a novel antagonistic human monoclonal antibody with a long VHCDR3 targeting the Class A GPCR formyl-peptide receptor 1

Contact Info

Product

Resources

About

Affinity maturation of a novel antagonistic human monoclonal antibody with a long V_HCDR3 targeting the Class A GPCR formyl-peptide receptor 1