scholarly journals Isolation and Characterization of Human Monoclonal Antibodies from Individuals Infected with West Nile Virus

2006 ◽  
Vol 80 (14) ◽  
pp. 6982-6992 ◽  
Author(s):  
Mark Throsby ◽  
Cecile Geuijen ◽  
Jaap Goudsmit ◽  
Arjen Q. Bakker ◽  
Jehanara Korimbocus ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
West Nile Virus ◽  
Viral Envelope ◽  
Phage Library ◽  
Enzyme Linked Immunosorbent Assay ◽  
West Nile ◽  
Isolation And Characterization ◽  
E Protein ◽  
Domain Iii

ABSTRACT Monoclonal antibodies (MAbs) neutralizing West Nile Virus (WNV) have been shown to protect against infection in animal models and have been identified as a correlate of protection in WNV vaccine studies. In the present study, antibody repertoires from three convalescent WNV-infected patients were cloned into an scFv phage library, and 138 human MAbs binding to WNV were identified. One hundred twenty-one MAbs specifically bound to the viral envelope (E) protein and four MAbs to the premembrane (prM) protein. Enzyme-linked immunosorbent assay-based competitive-binding assays with representative E protein-specific MAbs demonstrated that 24/51 (47%) bound to domain II while only 4/51 (8%) targeted domain III. In vitro neutralizing activity was demonstrated for 12 MAbs, and two of these, CR4374 and CR4353, protected mice from lethal WNV challenge at 50% protective doses of 12.9 and 357 μg/kg of body weight, respectively. Our data analyzing three infected individuals suggest that the human anti-WNV repertoire after natural infection is dominated by nonneutralizing or weakly neutralizing MAbs binding to domain II of the E protein, while domain III-binding MAbs able to potently neutralize WNV in vitro and in vivo are rare.

scholarly journals Recovery of West Nile Virus Envelope Protein Domain III Chimeras with Altered Antigenicity and Mouse Virulence

2016 ◽  
Vol 90 (9) ◽  
pp. 4757-4770 ◽  
Author(s):  
Alexander J. McAuley ◽  
Maricela Torres ◽  
Jessica A. Plante ◽  
Claire Y.-H. Huang ◽  
Dennis A. Bente ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Receptor Binding ◽  
Encephalitis Virus ◽  
Target Cells ◽  
West Nile ◽  
E Protein ◽  
Domain Iii ◽  
Chimeric Viruses

ABSTRACTFlaviviruses are positive-sense, single-stranded RNA viruses responsible for millions of human infections annually. The envelope (E) protein of flaviviruses comprises three structural domains, of which domain III (EIII) represents a discrete subunit. The EIII gene sequence typically encodes epitopes recognized by virus-specific, potently neutralizing antibodies, and EIII is believed to play a major role in receptor binding. In order to assess potential interactions between EIII and the remainder of the E protein and to assess the effects of EIII sequence substitutions on the antigenicity, growth, and virulence of a representative flavivirus, chimeric viruses were generated using the West Nile virus (WNV) infectious clone, into which EIIIs from nine flaviviruses with various levels of genetic diversity from WNV were substituted. Of the constructs tested, chimeras containing EIIIs from Koutango virus (KOUV), Japanese encephalitis virus (JEV), St. Louis encephalitis virus (SLEV), and Bagaza virus (BAGV) were successfully recovered. Characterization of the chimerasin vitroandin vivorevealed differences in growth and virulence between the viruses, within vivopathogenesis often not being correlated within vitrogrowth. Taken together, the data demonstrate that substitutions of EIII can allow the generation of viable chimeric viruses with significantly altered antigenicity and virulence.IMPORTANCEThe envelope (E) glycoprotein is the major protein present on the surface of flavivirus virions and is responsible for mediating virus binding and entry into target cells. Several viable West Nile virus (WNV) variants with chimeric E proteins in which the putative receptor-binding domain (EIII) sequences of other mosquito-borne flaviviruses were substituted in place of the WNV EIII were recovered, although the substitution of several more divergent EIII sequences was not tolerated. The differences in virulence and tissue tropism observed with the chimeric viruses indicate a significant role for this sequence in determining the pathogenesis of the virus within the mammalian host. Our studies demonstrate that these chimeras are viable and suggest that such recombinant viruses may be useful for investigation of domain-specific antibody responses and the more extensive definition of the contributions of EIII to the tropism and pathogenesis of WNV or other flaviviruses.

scholarly journals A VLP-based vaccine targeting domain III of the West Nile virus E protein protects from lethal infection in mice

2010 ◽  
Vol 7 (1) ◽  
Author(s):  
Gunther Spohn ◽  
Gary T Jennings ◽  
Byron EE Martina ◽  
Iris Keller ◽  
Markus Beck ◽  
...  
Keyword(s):  
West Nile Virus ◽  
The West ◽  
West Nile ◽  
E Protein ◽  
Lethal Infection ◽  
Domain Iii

Mosaic RNA Phage VLPs Carrying Domain III of the West Nile Virus E Protein

2014 ◽  
Vol 56 (5) ◽  
pp. 459-469 ◽  
Author(s):  
Indulis Cielens ◽  
Ludmila Jackevica ◽  
Arnis Strods ◽  
Andris Kazaks ◽  
Velta Ose ◽  
...  
Keyword(s):  
West Nile Virus ◽  
The West ◽  
West Nile ◽  
E Protein ◽  
Domain Iii ◽  
Rna Phage

scholarly journals Antiviral Peptides Targeting the West Nile Virus Envelope Protein

2006 ◽  
Vol 81 (4) ◽  
pp. 2047-2055 ◽  
Author(s):  
Fengwei Bai ◽  
Terrence Town ◽  
Deepti Pradhan ◽  
Jonathan Cox ◽  
Ashish ◽  
...  
Keyword(s):  
West Nile Virus ◽  
Envelope Protein ◽  
Phage Display Library ◽  
Viral Envelope ◽  
Host Cells ◽  
Viral Envelope Protein ◽  
Virus Envelope ◽  
West Nile ◽  
Inhibition Concentration

ABSTRACT West Nile virus (WNV) can cause fatal murine and human encephalitis. The viral envelope protein interacts with host cells. A murine brain cDNA phage display library was therefore probed with WNV envelope protein, resulting in the identification of several adherent peptides. Of these, peptide 1 prevented WNV infection in vitro with a 50% inhibition concentration of 67 μM and also inhibited infection of a related flavivirus, dengue virus. Peptide 9, a derivative of peptide 1, was a particularly potent inhibitor of WNV in vitro, with a 50% inhibition concentration of 2.6 μM. Moreover, mice challenged with WNV that had been incubated with peptide 9 had reduced viremia and fatality compared with control animals. Peptide 9 penetrated the murine blood-brain barrier and was found in the brain parenchyma, implying that it may have antiviral activity in the central nervous system. These short peptides serve as the basis for developing new therapeutics for West Nile encephalitis and, potentially, other flaviviruses.

scholarly journals Human Monoclonal Antibodies against West Nile Virus Induced by Natural Infection Neutralize at a Postattachment Step

2009 ◽  
Vol 83 (13) ◽  
pp. 6494-6507 ◽  
Author(s):  
Matthew R. Vogt ◽  
Bastiaan Moesker ◽  
Jaap Goudsmit ◽  
Mandy Jongeneelen ◽  
S. Kyle Austin ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
West Nile Virus ◽  
Cultured Cells ◽  
Natural Infection ◽  
Minor Component ◽  
Human Mabs ◽  
West Nile ◽  
Epidemic Encephalitis ◽  
E Protein

ABSTRACT West Nile virus (WNV) is a neurotropic flavivirus that is now a primary cause of epidemic encephalitis in North America. Studies of mice have demonstrated that the humoral immune response against WNV limits primary infection and protects against a secondary challenge. The most-potent neutralizing mouse monoclonal antibodies (MAbs) recognize an epitope on the lateral ridge of domain III (DIII-lr) of the envelope (E) protein. However, studies with serum from human patients show that antibodies against the DIII-lr epitope comprise, at best, a minor component of the human anti-WNV antibody response. Herein, we characterize in detail two WNV-specific human MAbs, CR4348 and CR4354, that were isolated from B-cell populations of convalescent patients. These MAbs strongly neutralize WNV infection of cultured cells, protect mice against lethal infection in vivo, and yet poorly recognize recombinant forms of the E protein. Instead, CR4348 and CR4354 bind determinants on intact WNV virions and subviral particles in a pH-sensitive manner, and neutralization is altered by mutations at the dimer interface in domain II and the hinge between domains I and II, respectively. CR4348 and CR4354 human MAbs neutralize infection at a postattachment step in the viral life cycle, likely by inhibiting acid-induced fusion within the endosome.

scholarly journals Development of Monoclonal Antibodies to West Nile Virus and Their Application in Immunohistochemistry

2012 ◽  
Vol 19 (11) ◽  
pp. 1853-1858 ◽  
Author(s):  
Jiro Hirota ◽  
Shinya Shimizu ◽  
Tomoyuki Shibahara ◽  
Takashi Isobe ◽  
Manabu Yamada ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
West Nile Virus ◽  
Japanese Encephalitis ◽  
Nonstructural Protein ◽  
Cross Reactivity ◽  
Encephalitis Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
West Nile ◽  
Nonstructural Protein 1 ◽  
Formalin Fixed Paraffin Embedded

ABSTRACTWest Nile virus (WNV) is endemic throughout Africa, Eurasia, America, and Australia and has important implications for avian, horse, and human health. In these regions, dead birds are monitored for the presence of WNV through immunohistochemistry (IHC) and PCR. However, a number of the tools for IHC are inadequate owing to their cross-reactivity to other Japanese encephalitis serogroup viruses. Here we have established eight monoclonal antibodies (MAbs) to WNV. Four of them bound to the envelope protein, three of them bound to nonstructural protein 1 (NS1), and one bound to precursor membrane protein (prM), as shown by Western blot analysis. The anti-NS1 MAbs and the anti-prM MAb did not cross-react with Japanese encephalitis virus (JEV), Murray valley encephalitis virus, or St. Louis encephalitis virus in an indirect enzyme-linked immunosorbent assay. One NS1-specific MAb, SHW-32B1, and the previously reported NS1-specific MAb, SHW-7A11, were shown by IHC to specifically detect the cytoplasm of degenerated cells in the heart and brain of a WNV-infected goose. Neither of these MAbs were shown by IHC to cross-react with degenerated cells in the brain of a JEV-infected pig. These MAbs are the first reported anti-NS1 MAbs that can be used for WNV-specific IHC using formalin-fixed, paraffin-embedded sections. They may be useful for WNV research and surveillance.

scholarly journals Development of a highly specific serodiagnostic ELISA for West Nile virus infection using subviral particles

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Keisuke Maezono ◽  
Shintaro Kobayashi ◽  
Koshiro Tabata ◽  
Kentaro Yoshii ◽  
Hiroaki Kariwa
Keyword(s):  
West Nile Virus ◽  
West Nile Virus Infection ◽  
Cross Reactivity ◽  
Terminal Region ◽  
Full Length ◽  
Enzyme Linked Immunosorbent Assay ◽  
E Proteins ◽  
West Nile ◽  
E Protein ◽  
Subviral Particles

AbstractWest Nile virus (WNV), a member of the Japanese encephalitis virus (JEV) serocomplex group, causes lethal encephalitis in humans and horses. Because serodiagnosis of WNV and JEV is hampered by cross-reactivity, the development of a simple, secure, and WNV-specific serodiagnostic system is required. The coexpression of prM protein and E protein leads to the secretion of subviral particles (SPs). Deletion of the C-terminal region of E protein is reported to affect the production of SPs by some flaviviruses. However, the influence of such a deletion on the properties and antigenicity of WNV E protein is unclear. We analyzed the properties of full-length E protein and E proteins lacking the C-terminal region as novel serodiagnostics for WNV infection. Deletion of the C-terminal region of E protein suppressed the formation of SPs but did not affect the production of E protein. The sensitivity of an enzyme-linked immunosorbent assay (ELISA) using the full-length E protein was higher than that using the truncated E proteins. Furthermore, in the ELISA using full-length E protein, there was little cross-reactivity with anti-JEV antibodies, and the sensitivity was similar to that of the neutralization test.

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