Induction of Epitope-Specific Neutralizing Antibodies against West Nile Virus

Oliphant, Theodore; Nybakken, Grant E.; Austin, S. Kyle; Xu, Qing; Bramson, Jonathan L.; Loeb, Mark; Throsby, Mark; Fremont, Daved H.; Pierson, Theodore C.; Diamond, Michael S.

doi:10.1128/jvi.00643-07

Cited by 161 publications

(216 citation statements)

References 61 publications

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“…The WNV E ectodomain (residues 1-404) and DIII (residues 296-404) bacterial expression constructs have been described (19). Proteins were expressed in E. coli and purified after an oxidative refolding protocol (19) and size-exclusion chromatography.…”

Section: Methodsmentioning

confidence: 99%

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Monoclonal antibody produced in plants efficiently treats West Nile virus infection in mice

Lai

Engle²,

Fuchs³

et al. 2010

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

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105

178

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Over the past decade, West Nile virus (WNV) has spread to all 48 of the lower United States as well as to parts of Canada, Mexico, the Caribbean, and South America, with outbreaks of neuroinvasive disease occurring annually. At present, no therapeutic or vaccine is available for human use. Epidemics of WNV and other emerging infectious disease threats demand cost-efficient and scalable production technologies that can rapidly transfer effective therapeutics into the clinical setting. We have previously reported that Hu-E16, a humanized anti-WNV mAb, binds to a highly conserved epitope on the envelope protein, blocks viral fusion, and shows promising postexposure therapeutic activity. Herein, we generated a plant-derived Hu-E16 mAb that can be rapidly scaled up for commercial production. Plant Hu-E16 was expressed at high levels within 8 days of infiltration in Nicotiana benthamiana plants and retained high-affinity binding and potent neutralizing activity in vitro against WNV. A single dose of plant Hu-E16 protected mice against WNV-induced mortality even 4 days after infection at rates that were indistinguishable from mammalian-cell-produced Hu-E16. This study demonstrates the efficacy of a plant-produced mAb against a potentially lethal infection several days after exposure in an animal challenge model and provides a proof of principle for the development of plant-derived mAbs as therapy against emerging infectious diseases.

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Section: Methodsmentioning

confidence: 99%

“…The ELISA for examining the binding of pHu-E16 to WNV E DIII was performed as described (19). DIII (amino acids 296-415) of the New York 1999 strain of WNV purified from Escherichia coli (19) was immobilized on microtiter plates.…”

Section: Methodsmentioning

confidence: 99%

Monoclonal antibody produced in plants efficiently treats West Nile virus infection in mice

Lai

Engle²,

Fuchs³

et al. 2010

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

Self Cite

105

178

View full text Add to dashboard Cite

show abstract

“…In this study, we administered IVIG at a high dose (1 g kg 21 ) to determine whether its antiinflammatory and immunomodulatory activities could protect against lethal WNV encephalitis, and we additionally tested IVIG adsorbed to be free of neutralizing antibodies directed to the envelope glycoprotein (E-gp). Most of the antibodies mediating protection against WNV infection are directed to E-gp (Oliphant et al, 2006(Oliphant et al, , 2007; hence, it was used to deplete IVIG of neutralizing antibodies. We found that a single dose of IVIG or WNV convalescent sera (WNV-IVIG) protected all mice from lethal WNV encephalitis when administered 24 h p.i.…”

Section: Introductionmentioning

confidence: 99%

Anti-inflammatory activity of intravenous immunoglobulins protects against West Nile virus encephalitis

Srivastava

Ramakrishna

Cantin

2015

Journal of General Virology

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West Nile virus (WNV), an important global human pathogen, targets neurons to cause lethal encephalitis, primarily in elderly and immunocompromised patients. Currently, there are no approved therapeutic agents or vaccines to treat WNV encephalitis. Recent studies have suggested that inflammation is a major contributor to WNV encephalitis morbidity. In this study we evaluated the use of IVIG (intravenous immunoglobulins -a clinical product comprising pooled human IgG) as an anti-inflammatory treatment in a model of lethal WNV infection. We report here that IVIG and pooled human WNV convalescent sera (WNV-IVIG) inhibited development of lethal WNV encephalitis by suppressing central nervous system (CNS) infiltration by CD45 high leukocytes. Pathogenic Ly6C high CD11b + monocytes were the major infiltrating subset in the CNS of infected control mice, whereas IVIG profoundly reduced infiltration of these pathogenic Ly6C high monocytes into the CNS of infected mice. Interestingly, WNV-IVIG was more efficacious than IVIG in controlling CNS inflammation when mice were challenged with a high-dose inoculum (10 5 versus 10 4 p.f.u.) of WNV. Importantly, adsorption of WNV E-glycoprotein neutralizing antibodies did not abrogate IVIG protection, consistent with virus neutralization not being essential for IVIG protection. These findings confirmed the potent immunomodulatory activity of generic IVIG, and emphasized its potential as an effective immunotherapeutic drug for encephalitis and other virus induced inflammatory diseases.

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“…By binding to the viral surface protein, antibodies can interfere with receptor attachment, virus internalization, or membrane fusion. Although the most potent inhibitory mouse antibodies against WNV bind to DIII of E (17)(18)(19), these appear to play a less dominant role in the human immune response against WNV infection (20). CR4354 is a strongly neutralizing human monoclonal antibody (MAb) against WNV that was isolated from a patient infected with WNV and inhibits infection subsequent to cell attachment (21).…”

mentioning

confidence: 99%

Neutralization of West Nile virus by cross-linking of its surface proteins with Fab fragments of the human monoclonal antibody CR4354

Kaufmann

Vogt²,

Goudsmit³

et al. 2010

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

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123

121

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Many flaviviruses are significant human pathogens, with the humoral immune response playing an essential role in restricting infection and disease. CR4354, a human monoclonal antibody isolated from a patient, neutralizes West Nile virus (WNV) infection at a postattachment stage in the viral life-cycle. Here, we determined the structure of WNV complexed with Fab fragments of CR4354 using cryoelectron microscopy. The outer glycoprotein shell of a mature WNV particle is formed by 30 rafts of three homodimers of the viral surface protein E. CR4354 binds to a discontinuous epitope formed by protein segments from two neighboring E molecules, but does not cause any detectable structural disturbance on the viral surface. The epitope occurs at two independent positions within an icosahedral asymmetric unit, resulting in 120 binding sites on the viral surface. The cross-linking of the six E monomers within one raft by four CR4354 Fab fragments suggests that the antibody neutralizes WNV by blocking the pH-induced rearrangement of the E protein required for virus fusion with the endosomal membrane.antibody | cryoelectron microscopy | flavivirus W est Nile virus (WNV) is a human pathogen that causes a febrile illness, which can progress to encephalitis, paralysis, and death. The virus is endemic in parts of Africa, Asia, and Europe, and in the past decade has spread throughout North America and into Central and South America (1). WNV is closely related to other arthropod-transmitted, medically relevant flaviviruses, such as dengue, yellow fever, Japanese encephalitis, and tick-borne encephalitis viruses. These lipid-enveloped viruses enter host cells by receptor-mediated endocytosis. The singlestranded, positive-sense RNA genome is released into the cytoplasm after low pH induces the fusion of the viral lipid envelope with the endosomal membrane.Mature WNV virions are roughly spherical with a diameter of about 500 Å. The outer viral surface is composed of an icosahedral scaffold of 180 closely packed copies of the membrane-anchored envelope (E) glycoprotein. Sets of three, nearly parallel E homodimers are associated into rafts that form a herringbone pattern on the surface of mature virions. The ectodomain of E has three structural domains, DI, DII, and DIII (2-5), with domain DI positioned structurally between DII and DIII. DII contains a fusion loop at its distal end that is indispensable for virus-cell membrane fusion. The Ig-like C-terminal domain DIII undergoes a major, pH-triggered positional rearrangement essential for fusion, and may also be involved in receptor binding (2, 6-12). During cell entry of flaviviruses, low endosomal pH triggers a proposed E protein rearrangement cascade, including the dissociation of E dimers and the outward rotation of DII during the repositioning of E monomers into fusion-active trimers (6, 9, 13).The humoral immune response is essential for protection against flavivirus infection and disease (14,15). The E glycoprotein is the principal antigen that elicits neutralizing antibodies agai...

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Induction of Epitope-Specific Neutralizing Antibodies against West Nile Virus

Cited by 161 publications

References 61 publications

Monoclonal antibody produced in plants efficiently treats West Nile virus infection in mice

Monoclonal antibody produced in plants efficiently treats West Nile virus infection in mice

Anti-inflammatory activity of intravenous immunoglobulins protects against West Nile virus encephalitis

Neutralization of West Nile virus by cross-linking of its surface proteins with Fab fragments of the human monoclonal antibody CR4354

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