Mechanism of Dengue Virus Broad Cross-Neutralization by a Monoclonal Antibody

Cockburn, J.J.B.; Sanchez, M.E. Navarro; Fretes, N.; Urvoas, Agathe; Staropoli, Isabelle; Kikuti, Carlos; Coffey, Lark L.; Seisdedos, F. Arenzana; Bedouelle, Hugues; Rey, F.A.

doi:10.1016/j.str.2012.01.001

Cited by 126 publications

(178 citation statements)

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“…Two mouse-adapted strains (DENV-2 and DENV-3) have been reported to cause lethal infection in wild-type mice (82)(83)(84), although mechanistically how this occurs remains unknown, as DENV isolates uniformly fail to an- a Amino acids were determined based on loss of binding to the indicated MAb by using prM-E display of a shotgun alanine-scanning mutagenesis library (see Materials and Methods for details). b Loops and strands that comprised specific epitopes were assigned based on existing crystal structures of DIII from DENV-1, DENV-2, and DENV-3 E proteins (17,19,20,89).…”

Section: Discussionmentioning

confidence: 99%

Functional Analysis of Antibodies against Dengue Virus Type 4 Reveals Strain-Dependent Epitope Exposure That Impacts Neutralization and Protection

Sukupolvi-Petty

Brien

Austin

et al. 2013

J Virol

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g Although prior studies have characterized the neutralizing activities of monoclonal antibodies (MAbs) against dengue virus (DENV) serotypes 1, 2, and 3 (DENV-1, DENV-2, and DENV-3), few reports have assessed the activity of MAbs against DENV-4. Here, we evaluated the inhibitory activity of 81 new mouse anti-DENV-4 MAbs. We observed strain-and genotype-dependent differences in neutralization of DENV-4 by MAbs mapping to epitopes on domain II (DII) and DIII of the envelope (E) protein. Several anti-DENV-4 MAbs inefficiently inhibited at least one strain and/or genotype, suggesting that the exposure or sequence of neutralizing epitopes varies within isolates of this serotype. Remarkably, flavivirus cross-reactive MAbs, which bound to the highly conserved fusion loop in DII and inhibited infection of DENV-1, DENV-2, and DENV-3, more weakly neutralized five different DENV-4 strains encompassing the genetic diversity of the serotype after preincubation at 37°C. However, increasing the time of preincubation at 37°C or raising the temperature to 40°C enhanced the potency of DII fusion loop-specific MAbs and some DIII-specific MAbs against DENV-4 strains. Prophylaxis studies in two new DENV-4 mouse models showed that neutralization titers of MAbs after preincubation at 37°C correlated with activity in vivo. Our studies establish the complexity of MAb recognition against DENV-4 and suggest that differences in epitope exposure relative to other DENV serotypes affect antibody neutralization and protective activity. Dengue virus (DENV) is a member of the Flaviviridae family of RNA viruses and is genetically related to other human pathogens of global concern, including yellow fever, West Nile (WNV), and Japanese encephalitis viruses. In nature, DENV disease occurs exclusively in humans after Aedes aegypti or Aedes albopictus mosquito inoculation, with clinical disease ranging from a debilitating febrile illness (dengue fever [DF]) to a life-threatening hemorrhagic and capillary leak syndrome (dengue hemorrhagic fever [DHF]/dengue shock syndrome [DSS]). No approved antiviral treatment is currently available, although candidate tetravalent vaccines are in advanced clinical trials (reviewed in references 1 and 2). Because of the increased geographic range of its mosquito vectors, urbanization, and international travel, DENV continues to spread worldwide and now causes an estimated 390 million infections and 500,000 cases of DHF/DSS per year, with 3.6 billion people at risk (3, 4). Given that the most advanced live-attenuated DENV vaccine candidate showed a poor 30% overall efficacy rate in a recently published phase 2b clinical trial (5), there is an urgent need for understanding the correlates of protection, especially those of neutralizing antibodies.DENV is an enveloped virus with a single-stranded, positivepolarity RNA genome. Based on experiments with DENV-1 and DENV-2 isolates, the mature DENV virion is ϳ500 Å in diameter with a highly organized outer protein shell, a 50-Å lipid membrane bilayer, and a nucleocapsid core ...

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Section: Discussionmentioning

confidence: 99%

Functional Analysis of Antibodies against Dengue Virus Type 4 Reveals Strain-Dependent Epitope Exposure That Impacts Neutralization and Protection

Sukupolvi-Petty

Brien

Austin

et al. 2013

J Virol

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“…This is relevant because a previous structure of DENV--4 sE in complex with an antibody that binds away from the EDE also had the 150 loop disordered 13 , highlighting an intrinsic mobility in this area depending on the E amino acid sequence. Displacement of the 150 loop allows the EDE1 light chain to come closer to sE and interact with domain III (compare panel e with f and g in Fig.1) in the region of the "A strand" epitope, which has been structurally characterized previously for murine DENV cross--reactive antibodies 16,17 . These domain III contacts are centered on the conserved E residue K310, the side chain of which makes a lid covering the indole ring of W101 of the fusion loop (ED Fig.…”

mentioning

confidence: 89%

Recognition determinants of broadly neutralizing human antibodies against dengue viruses

Rouvinski

Guardado-Calvo

Barba-Spaeth

et al. 2015

Nature

309

353

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2Dengue disease is caused by four different flavivirus 1 serotypes, which infect 390 million people yearly with 25% symptomatic cases 2 and for which no licensed vaccine is available. Recent phase III vaccine trials showed partial protection, and in particular no protection for dengue virus serotype 2 (DENV--2) 3,4 . Structural studies so far have characterized only epitopes recognized by serotype specific human antibodies 5,6 . We recently isolated human antibodies potently neutralizing all four DENV serotypes 7 . Here we describe the X--ray structures of four of these broadly neutralizing antibodies (bnAbs) in complex with the envelope glycoprotein E from DENV--2, revealing that the recognition determinants are at a serotype conserved site at the E dimer interface, including the exposed main chain of the E fusion loop 8 and the two conserved glycan chains.This "E--dimer dependent epitope" (EDE) is also the binding site for the viral glycoprotein prM during virus maturation in the secretory pathway of the infected cell 9 , explaining its conservation across serotypes and highlighting an Achilles heel of the virus with respect to antibody neutralization. These findings will be instrumental for devising novel immunogens to protect simultaneously against all four serotypes of dengue virus.Exposed at the surface of infectious mature DENV particles, protein E is the sole target of neutralizing antibodies. It displays an icosahedral arrangement in which 90 E dimers completely coat the viral surface 10,11 and which is sensitive to the environmental pH. Upon entry of DENV into cells via receptor--mediated endocytosis, the acidic 3 endosomal environment triggers an irreversible fusogenic conformational change in E that leads to fusion of viral and endosomal membranes 1 . The structure of the isolated E dimer has been determined by X--ray crystallography using the soluble ectodomain (sE) 8,12 . Protein E is relatively conserved, displaying about 65% amino acid sequence identity when comparing the most distant DENV serotypes. In particular, there are two conserved N--linked glycosylation sites at positions N67 and N153. To examine its interaction with the antibodies, we selected four highly potent bnAbs identified in the accompanying work: 747(4) A11 and 747 B7 (EDE2 group, requiring glycosylation at position N153 for efficient binding) and 752--2 C8 and 753(3) C10 (EDE1 group, binding regardless of the glycosylation at N153) 7 -referred to as A11, B7, C8 and C10 from hereon. The EDE2 bnAbs were isolated from the same patient (who had a secondary infection with DENV--2), and are somatic variants of the same IgG clone, derived from the IGHV3--74 and IGLV2--23 germ lines. The heavy chain has a very long (26 amino acids, IMGT convention) complementarity--determining region 3 (CDR H3). The EDE1 bnAbs were isolated from different patients and derive from (EDE1 C8, the patient appeared to have a primary infection of undetermined serotype) and IGHV1--3* and IGLV2--14 (EDE1 C10, from a patient with secondary DENV--1 infecti...

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“…Ultimately no infectious virus was detected in this or in subsequent C6/36 passages. These results imply an important structural role for residue K310, which forms a latch on a hydrophobic pocket containing the fusion loop and is also involved in inter-dimer interactions (Cockburn et al, 2012). The decreased mass and lack of charge of alanine may expose the hydrophobic pocket and also prevent the inter-dimer interactions required for infectious virus ( Fig.…”

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confidence: 91%

Functional analysis of dengue virus (DENV) type 2 envelope protein domain 3 type-specific and DENV complex-reactive critical epitope residues

Pitcher

Sarathy

Matsui

et al. 2015

Journal of General Virology

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The dengue virus (DENV) envelope protein domain 3 (ED3) is the target of potent virus neutralizing antibodies. The DENV-2 ED3 contains adjacent type-specific and DENV complex-reactive antigenic sites that are composed of a small number of residues that were previously demonstrated to be critical for antibody binding. Site-directed mutagenesis of a DENV-2 16681 infectious clone was used to mutate critical residues in the DENV-2 type-specific (K305A and P384A) and DENV complex-reactive (K310A) antigenic sites. The K305A mutant virus multiplied like the parent virus in mosquito and mammalian cells, as did the P384A mutant virus, which required a compensatory mutation (G330D) for viability. However, the K310A mutant virus could not be recovered. The DENV-2 type-specific critical residue mutations K305A and P384A+G330D reduced the ability of DENV-2 type-specific, but not DENV complex-reactive, mAbs to neutralize virus infectivity and this was directly correlated with mAb binding affinity to the rED3 mutants.The disease dengue (DEN) is caused by four serologically and genetically related dengue viruses (DENVs) termed DENV-1, -2, -3 and -4. The DENV envelope (E) protein is composed of three domains: E protein domain I (ED1) is the central domain, ED2 is the dimerization domain and contains the conserved fusion loop, and ED3 is the putative receptor-binding domain (Modis et al

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Mechanism of Dengue Virus Broad Cross-Neutralization by a Monoclonal Antibody

Cited by 126 publications

References 48 publications

Functional Analysis of Antibodies against Dengue Virus Type 4 Reveals Strain-Dependent Epitope Exposure That Impacts Neutralization and Protection

Functional Analysis of Antibodies against Dengue Virus Type 4 Reveals Strain-Dependent Epitope Exposure That Impacts Neutralization and Protection

Recognition determinants of broadly neutralizing human antibodies against dengue viruses

Functional analysis of dengue virus (DENV) type 2 envelope protein domain 3 type-specific and DENV complex-reactive critical epitope residues

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