Anti-Human Immunodeficiency Virus Type 1 (HIV-1) Antibodies 2F5 and 4E10 Require Surprisingly Few Crucial Residues in the Membrane-Proximal External Region of Glycoprotein gp41 To Neutralize HIV-1

Zwick, Michael B.; Jensen, Richard Christian; Church, S.; Wang, Meng; Stiegler, Gabriela; Kunert, Renate; Katinger, Hermann; Burton, Dennis R.

doi:10.1128/jvi.79.2.1252-1261.2005

Cited by 261 publications

(304 citation statements)

References 79 publications

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“…HIV-1 Env pseudotyped virus clones containing L669 alanine or serine mutations with enhanced sensitivity to MPER neutralizing antibodies have been previously reported (5,20). For this study, we used env-pseudotyped viruses with the wild-type 669L (termed Env TND_669L pseudotyped virus) and the Env pseudotyped virus TND_669S with the 669S mutation (Fig.…”

Section: A Single L669s Mutation Accounts For the Enhanced Potency Ofmentioning

confidence: 99%

Prolonged exposure of the HIV-1 gp41 membrane proximal region with L669S substitution

Shen

Dennison

Liu

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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The conserved membrane-proximal external region (MPER) of HIV-1 envelope is a target for the rare broadly neutralizing 2F5, Z13, and 4E10 monoclonal antibodies (mAbs). One strategy to elicit such antibodies is to design an immunogen with increased exposure of the 2F5 and 4E10 mAb epitopes. In this study we characterize a single leucine to serine substitution at position 669 (L669S) in the gp41 Env MPER that confers >250-fold more neutralization sensitivity to 2F5 and 4E10 mAbs than does the wild-type gp41 sequence. On synthetic liposomes, increased solvent exposure of MPER tryptophan residues and stable docking of 2F5 and 4E10 mAbs to mutant MPER peptide liposomes indicate more favorable membrane orientation of MPER neutralizing epitopes with L669S substitution. The time during which virus is sensitive to 2F5 mAb-mediated neutralization is approximately 3-fold longer when the mutation is present. These data suggest that a major contribution to the L669S mutant virus phenotype of enhanced susceptibility to MPER mAbs is prolonged exposure of the MPER neutralizing epitope during viral entry. immunogen | broadly neutralizing antibodiesA major challenge for an effective HIV-1 vaccine is the inability of immunogens to induce broadly neutralizing antibodies (nAbs). One goal for antibody-based HIV-1 vaccine strategies is to elicit broadly neutralizing antibodies similar in breadth to the membrane-proximal external region (MPER) 2F5 and 4E10 monoclonal antibodies (mAbs) that neutralize a majority of transmitted viruses (1). MPER-specific broadly neutralizing antibodies are rarely made in HIV-1 infection (2-4), but recent studies from our group and others have shown that 2F5-like antibodies responsible for neutralization breadth can be found in ≈0.3% of HIV-1 infected subjects (5), whereas 4E10-like antibodies can be found in ∼3% of HIV-1 positive subjects (6). Vaccination with antigenic envelope constructs expressing 2F5 and 4E10 epitopes has not induced high-titered neutralizing antibodies (7-10). One hypothesis for the failure of such vaccines to elicit broadly neutralizing antibodies is that the Env epitopes presented to host B cells are not in the correct envelope conformation; for the MPER, this conformation may be the transient, prehairpin gp41 intermediate (11,12). Another hypothesis is that 2F5 and 4E10 mAbs, which are unusual in having long hydrophobic CDR3 loops, may be particularly effective in reaching epitopes near the virion lipid bilayer, but may be difficult to induce because of down-regulation of polyreactive B cell clones through B cell tolerance mechanisms (13-18).Previous work has identified two amino acid substitutions (T569A, I675V) (19) and L669S (5) in the MPER that increased neutralization sensitivity by MPER antibodies; however, the mechanism by which neutralization sensitivity was increased was not determined (5,19). We now report that the mechanism of enhanced neutralization sensitivity of a single amino acid substitution (leucine to serine substitution at position 669, L669S) in the heptad re...

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Section: A Single L669s Mutation Accounts For the Enhanced Potency Ofmentioning

confidence: 99%

Prolonged exposure of the HIV-1 gp41 membrane proximal region with L669S substitution

Shen

Dennison

Liu

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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“…Similarly, humans treated with T20/Fuzeon make high levels of anti-T20 antibody that does not inhibit T20 efficacy as a fusion inhibitor [123]. Interestingly a mAb, D50, that binds to an epitope just N-terminal to the 2F5 epitope, also binds to T20, but does not inhibit T20 activity as a fusion inhibitor [124,125]. One possible explanation for the poor immunogenicity of the MPER neutralizing epitopes is that the conformation of the gp41 MPER region in most immunogens is not in the correct prefusion confirmation to induce 2F5 and 4E10-like antibodies, but rather is in a non-native conformation.…”

mentioning

confidence: 99%

Aiming to induce broadly reactive neutralizing antibody responses with HIV-1 vaccine candidates

Haynes¹,

Montefiori²

2006

Expert Review of Vaccines

103

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Neutralizing antibody induction is a key feature of many effective vaccines and is the only immune response that has proven to be capable of completely blocking AIDS virus infection in animal models. Unfortunately, the extensive genetic variability and complex immune-evasion strategies of HIV-1 have thwarted all attempts to date at eliciting an effective neutralizing antibody response with candidate HIV-1 vaccine immunogens. Recent advances in our understanding of how these evasion strategies operate, coupled with growing progress in unravelling the structure and immunobiology of the viral envelope glycoproteins, are contributing to novel immunogen designs to overcome the many barriers to inducing protective antibodies against HIV-1. Keywordsantiretroviral therapy; HIV-1; host cell fusion; immunogen; neutralizing antibody induction Development of a safe, practical and effective HIV-1 vaccine is one of the highest priorities of the global scientific community [1,2]. Although antiretroviral therapy (ART) has dramatically prolonged the lives of HIV-1 infected patients, ART is not yet routinely available in developing countries, and the global rate of spread of HIV-1 continues unabated. If no effective AIDS vaccine is developed by 2010, the number of people infected world-wide with HIV-1 could exceed 60 million [3].In spite of more than 20 years of research, the types of immune responses needed to protect an immunized individual from HIV-1 infection are not known. Its is known that CD8 + cytotoxic T-cell responses can control HIV-1 replication to varying degrees in acute HIV-1 infection (AHI) [4,5], and when induced by immunogens, can control viral set point after simian immunodeficiancy virus (SIV) or simian HIV (SHIV) challenges in non-human primates [4]. Strong proliferative CD4 + T-cell responses to HIV-1 proteins have been demonstrated to correlate well with immune control of HIV-1 viral load [5]. Therefore, it is highly desirable for an HIV-1 vaccine co induce robust CD4 + and CD8 + T-cell responses in order to control virus replication and reduce the likelihood of subsequent transmission [4][5][6][7]. The potential for complete ('sterilizing') immunity from HIV-1 infection may depend on the presence of pre-existing neutralizing antibodies. We know that neutralizing antibodies can prevent the acquisition of AIDS virus infection after intravenous, vaginal, rectal and oral virus challenge in nonhuman primates [8][9][10][11][12]. This level of protection is highly attractive for a vaccine against a virus, such as HIV-1, which integrates genetically and forms latent viral reservoirs soon after infection. However, the diversity of transmitted HIV-1 genetic variants and the relative resistance of the majority of HIV-1 primary isolates to most types of inducible neutralizing antibodies have posed major hurdles to current vaccine efforts. Furthermore, the few rare broadly reactive neutralizing monoclonal antibodies (mAbs) that have been isolated from HIV-1 infected patients represent species of antibodies that...

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“…1). These core epitopes were determined by peptide mapping; however, alanine-scanning mutagenesis of the MPER in intact viruses established that only a few residues (D664/K665/W666, W672/F673/W680, and N671/ D674 HXB2 numbering throughout) are essential for 2F5, 4E10, and Z13e1 neutralization, respectively (7,9). Crystal structures of Fab 2F5, 4E10, and Z13e1 complexed with their respective epitope peptides subsequently confirmed that residues DKW, WFW, and ND are buried in the paratopes of these antibodies (10)(11)(12)(13)(14).…”

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confidence: 99%

Aromatic residues at the edge of the antibody combining site facilitate viral glycoprotein recognition through membrane interactions

Scherer

Leaman

Zwick

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

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107

117

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The broadly neutralizing anti-HIV antibody 4E10 recognizes an epitope very close to the virus membrane on the glycoprotein gp41. It was previously shown that epitope recognition improves in a membrane context and that 4E10 binds directly, albeit weakly, to lipids. Furthermore, a crystal structure of Fab 4E10 complexed to an epitope peptide revealed that the centrally placed, protruding H3 loop of the antibody heavy chain does not form peptide contacts. To investigate the hypothesis that the H3 loop apex might interact with the viral membrane, two Trp residues in this region were substituted separately or in combination with either Ala or Asp by site-directed mutagenesis. The resultant IgG variants exhibited similar affinities for an epitope peptide as WT 4E10 but lower apparent affinities for both viral membrane mimetic liposomes and Env(−) virus. Variants also exhibited lower apparent affinities for Env(+) virions and failed to significantly neutralize a number of 4E10-sensitive viruses. For the extremely sensitive HXB2 virus, variants did neutralize, but at 37-to >250-fold lower titers than WT 4E10, with Asp substitutions exerting a greater effect on neutralization potency than Ala substitutions. Because reductions in lipid binding reflect trends in neutralization potency, we conclude that Trp residues in the antibody H3 loop enable membrane proximal epitope recognition through favorable lipid interactions. The requirement for lipophilic residues such as Trp adjacent to the antigen binding site may explain difficulties in eliciting 4E10-like neutralizing antibody responses by immunization and helps define a unique motif for antibody recognition of membrane proximal antigens.4E10 | gp41 | HIV | neutralizing antibody | tryptophan

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Anti-Human Immunodeficiency Virus Type 1 (HIV-1) Antibodies 2F5 and 4E10 Require Surprisingly Few Crucial Residues in the Membrane-Proximal External Region of Glycoprotein gp41 To Neutralize HIV-1

Cited by 261 publications

References 79 publications

Prolonged exposure of the HIV-1 gp41 membrane proximal region with L669S substitution

Prolonged exposure of the HIV-1 gp41 membrane proximal region with L669S substitution

Aiming to induce broadly reactive neutralizing antibody responses with HIV-1 vaccine candidates

Aromatic residues at the edge of the antibody combining site facilitate viral glycoprotein recognition through membrane interactions

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